bromochloroacetic-acid and Lung-Neoplasms

Circulating tumour cells (CTCs) are a rare cell subpopulation regulated by the tumour microenvironment. In hypoxic conditions, CTCs are able to invade the lymphatic and circulatory systems leading to metastasis at distant sites.. To mimic in vivo oxygen variations and effects on CTCs, we have cultured five non-small cell lung cancer (NSCLC) cell lines under normoxic and hypoxic conditions, followed by a pulse of reoxygenation for 4 h.. Proliferation, spheroid-formation and colony formation ability under varying O. Our data suggest that when investigating CTCs as a prognostic biomarker in NSCLC, it is also essential to take into consideration EMT status to obtain a comprehensive overview of CTCs in circulation.

Topics: Biomarkers; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cell Line; Cell Line, Tumor; Epithelial-Mesenchymal Transition; ErbB Receptors; Humans; Hypoxia; Keratins; Lung Neoplasms; Neoplastic Cells, Circulating; Oxygen; RNA, Messenger; Tumor Microenvironment; Vimentin

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; Carcinoma, Renal Cell; Diagnosis, Differential; Humans; Keratins; Kidney Neoplasms; Liver Neoplasms; Lung Neoplasms; Male; Neoplasm Proteins; Neoplasms, Second Primary; Neoplasms, Unknown Primary; Organ Specificity; Prostatic Neoplasms

Hyalinizing clear cell carcinoma (HCCC) is common in head and neck sites but extremely rare in the lung. This case report describes an HCCC in the lung of a 54-year-old female patient.. We summarize the histomorphologic, immunophenotypic, and molecular features for our and three previously reported HCCCs of the lung with emphasis on potential diagnostic pitfalls.. Sections of a well-circumscribed 3.5-cm lung mass were characterized by a bronchocentric tumor growing in sheets, nests, and cords in a background of hyalinized stroma. Tumor cell appearance was clear to eosinophilic, lacking significant pleomorphism or mitotic activity. By immunohistochemistry, the tumor cells were strongly positive with antibodies to pan-keratin, p63, and CK5/6 while negative for CK7, CK20, thyroid transcription factor 1, napsin A, chromogranin, and synaptophysin. Next-generation sequencing demonstrated an EWSR1-ATF1 fusion transcript.. Awareness of key morphologic features of pulmonary HCCC is crucial for the recognition of this rare entity in the lung. Ancillary studies, including immunohistochemistry and molecular testing, are essential for the distinction from its mimics.

Topics: Adenocarcinoma, Clear Cell; Biomarkers, Tumor; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Membrane Proteins; Middle Aged; Oncogene Proteins, Fusion

The diagnosis of large cell carcinoma can only be made on a sampled resected tumor and should not be applied to biopsies or cytology. In the 2015 WHO classification, the definition of large cell carcinoma is restricted to carcinomas both lacking morphological signs of glandular, squamous or neuroendocrine differentiation and exhibiting a null or unclear phenotype (TTF1-/p40 ou p63 ou CK5/6+ focally). These carcinomas have an adenocarcinoma molecular profile because they harbor a significant number of KRAS and BRAF mutations, a profile that is more similar to adenocarcinoma than squamous cell carcinoma. They also have a worse prognosis than the other types of non-small cell lung carcinoma. Many large cell carcinomas previously classified on morphological data alone are now reclassified in the adenocarcinoma and squamous cell carcinoma types, including immunohistochemical features. The other large cell carcinoma subtypes from the 2004 WHO classification, i.e. large cell neuroendocrine carcinoma and basaloid carcinoma, are grouped respectively with the other neuroendocrine tumors and squamous cell carcinomas. Clear cell and rhabdoid features are now considered as cytological variants that can occur in any histopathological subtype and not as distinct subtypes. Lymphoepithelioma-like carcinoma is moved to the group of other and unclassified carcinomas as NUT carcinoma.

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoma, Large Cell; Cell Differentiation; DNA-Binding Proteins; Genes, ras; Humans; Immunophenotyping; Keratins; Lung Neoplasms; Prognosis; Proto-Oncogene Proteins B-raf; Transcription Factors; Tumor Suppressor Proteins

The functional role of IKKα in vivo is pretty complicated, largely due to its diverse functions through cell autonomous and non-autonomous manners. In addition, most of the studies on IKKα were derived from animal models, whether these findings hold true in human tumors remain unclear. Here we examined the expression of IKKα in nasopharyngeal carcinoma, which includes non-keratinizing carcinoma and keratinizing squamous cell carcinoma, and lung squamous cell carcinoma with keratinization and non-keratinization. We demonstrated that IKKα expression was almost negative in keratinizing cancer and higher expression of IKKα was found in non-keratinizing cancer, and that IKKα expression correlated with cellular differentiation of tumors in non-keratinizing nasopharyngeal carcinoma. These findings demonstrate that IKKα is diversely expressed in keratinizing and non-keratinizing carcinomas in the same type of cancer.

Topics: Biomarkers, Tumor; Carcinoma; Carcinoma, Squamous Cell; Cell Differentiation; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; I-kappa B Kinase; Kaplan-Meier Estimate; Keratins; Lung Neoplasms; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Phenotype; Prognosis; Time Factors

This report describes the clinical, histological and immunohistochemical features of two patients with primary pulmonary synovial sarcoma in the context of the literature. Chest pain, cough, haemoptysis and an enlarging pleural-based mass are the main clinical manifestations. Diagnosis depends on identifying epithelioid or spindle cells microscopically and on immunohistochemistry showing positivity for cytokeratin and vimentin and epithelial membrane antigen stains. Surgical excision is the main treatment approach.

Topics: Adult; Chest Pain; Cough; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Sarcoma, Synovial; Vimentin

Epithelioid hemangioendothelioma (EHE) is a rare tumor of vascular origin. While it can be found in any tissue, it is most often found in lung and liver and usually has an intermediate behavior. EHEs originating from pleural tissue have been less frequently described than those from other sites. Furthermore, to date, all of the cited pleural EHEs were described as highly aggressive. In the present report, we describe a rare case of pleural EHE extending to lung and bone in a 31-year-old woman. The histological diagnosis was confirmed by both conventional examination and immunohistochemistry. Her disease stabilized during the 4th course of adriamycin (45 mg/m(2), day 1-3), dacarbazine (300 mg/m(2), day 1-3) and ifosfamide (2,500 mg/m(2), day 1-3) with mesna, and she survived for 10 months after the diagnosis.

Topics: Adult; Antineoplastic Combined Chemotherapy Protocols; Diagnosis, Differential; Factor VIII; Female; Hemangioendothelioma, Epithelioid; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Pleural Neoplasms; Vimentin

To analyze the clinicopathological and immunohistochemical features of pulmonary blastoma (PB).. Four patients with PB, 1 male and 3 females, with the onset age of 2 months approximately 80 years, underwent resection of the tumor and were followed up for 32 months at most. The clinical data were analyzed retrospectively.. Pathology showed that the 4 cases all suffered from epithelial type PB. Under microscope, epithelial cells were atypical, mostly in mitogenic phase, and were lined up as many dense tubes. All of the patients underwent surgical resection and 3 - 5 cycles of chemotherapy and one had radiotherapy following surgery. The survival time was 5 - 32 months. One of the four patients died during follow-up due to metastasis 5 months after operation. The other patients still survived. Immunohistochemistry showed that cytokeratin and thyroid transcription factor-1 were positive, and vimentin, epithelial membrane antigen, and S-100 protein were negative in the tumor tissues; and a part of tumor cell presented positive Cg-A.. PB is rare and presents different clinical features. It is difficult to determine the diagnosis before operation. The modular structures and expression of neuroendocrine markers are helpful in differentiating epithelial type PB from usual adenocarcinoma in immunohistochemical staining.

Topics: Adult; Child; Disease-Free Survival; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Nuclear Proteins; Pulmonary Blastoma; Thyroid Nuclear Factor 1; Transcription Factors

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Adenocarcinoma, Mucinous; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Male; Middle Aged

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Humans; Keratins; Keratins, Type I; Keratins, Type II; Lung Neoplasms

Topics: Apoptosis; Breast Neoplasms; Drug Resistance, Neoplasm; Female; Gene Expression Regulation, Neoplastic; Humans; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Neoplasm Metastasis; Neoplasm Seeding; Neoplasms; Neoplastic Cells, Circulating

This is a case report of a rare patient with primary pulmonary synovial sarcoma. The patient was a 58-year-old woman who presented with a well-defined giant mass in the right lower field on a chest radiograph. A malignant pulmonary tumor was suspected and consequently a right middle and lower lobectomy was performed. Grossly, the tumor measured 10 x 8 x 7 cm, was whitish-yellow in color and friable with hemorrhage. Histologically, the tumor showed a dense proliferation of spindle cells. In some areas, a herringbone-like pattern with coagulation necrosis of large size was noted. Immunohistochemically, the tumor cells were focally positive for cytokeratin and epithelial membrane antigen (EMA). As these features suggested a monophasic synovial sarcoma, we looked for the presence of SYT-SSX fusion gene transcripts using RNA samples from the paraffin-embedded tissue. A reverse transcription-polymerase chain reaction (RT-PCR) amplified a single 118 bp fragment characteristic of the SYT-SSX1 fusion gene transcripts. As no tumor was found at other sites, it was diagnosed as primary pulmonary synovial sarcoma. Molecular testing proved to be very helpful or necessary when monophasic spindle cell synovial sarcoma was recognized in uncommon/unexpected sites. In our review of primary pulmonary synovial sarcomas confirmed by molecular detection of SYT-SSX fusion gene transcripts, the SYT-SSX2 fusion protein expression correlates with poorer prognosis. This is in contrast to the association between the SYT-SSX1 fusion protein expression and poorer prognosis in soft tissue synovial sarcomas.

Topics: Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Oncogene Proteins, Fusion; Paraffin Embedding; Reverse Transcriptase Polymerase Chain Reaction; Sarcoma, Synovial; Transcription, Genetic; Vimentin

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Large Cell; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Electrochemistry; Humans; Immunoenzyme Techniques; Keratin-19; Keratins; Luminescent Measurements; Lung Neoplasms; Radioimmunoassay; Reference Values; Specimen Handling

The purpose of this study was to determine the prognostic significance of a high pretreatment serum CYFRA 21-1 level (a cytokeratin 19 fragment) adjusted for the effects of well-known co-variables in non-small-cell lung cancer (NSCLC). This meta-analysis based on individual updated data gathered comprehensive databases from published or unpublished controlled studies dealing with the prognostic effect of serum CYFRA 21-1 level at presentation in NSCLC of any stage (nine institutions, 2063 patients). Multivariate regression was carried out with the Cox model. The proportional hazard assumption for each of the selected variables retained in the final model was originally checked by log minus log plots baseline hazard ratio. The follow-up ranged from 25 to 78 months. A total of 1616 events were recorded. In the multivariate analysis performed at the 1-year end point, a high pretreatment CYFRA 21-1 level was an unfavourable prognostic determinant in all centres except one (Hazard ratio (95% confidence interval): 1.88 (1.64-2.15), P<10(-4)). Other significant variables were stage of the disease, age and performance status. Within the first 18 months, the procedure disclosed a nearly similar hazard ratio for patients having a high pretreatment serum CYFRA 21-1 level (1.62 (1.42-1.86), P<10(-4)). For patients who did not undergo surgery, the hazard ratio during the first year of follow-up was 1.78 (1.54-2.07), P<10(-4). Finally, in the surgically treated population, at the 2-year end point, a high pretreatment CYFRA 21-1 and a locally advanced stage remained unfavourable prognostic determinants. In conclusion CYFRA 21-1 might be regarded as a putative co-variable in analysing NSCLC outcome inasmuch as a high serum level is a significant determinant of poor prognosis whatever the planned treatment.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Models, Theoretical; Neoplasm Staging; Patient Care Planning; Prognosis; Survival Analysis

Tumor markers are defined as substances which are produced by cancer cells or non-cancer cells reactive to cancer cells, and reflect the cancer status, such as its presence, characteristics, and volume. Clinically, many tumor markers are useful not only to assess the presence/absence of cancer, the primary site, histology, stage, and recurrence, but also to monitor the anti-cancer therapy. Tumor markers for lung cancer play only supporting roles because of their limited sensitivity and specificity, but they are clinically essential to daily medical oncology. This review addresses 6 important tumor markers for lung cancer, namely, CEA, SLX, CYFRA, SCC, ProGRP, and NSE.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Humans; Keratins; Lung Neoplasms; Oligosaccharides; Peptide Fragments; Peptides; Phosphopyruvate Hydratase; Recombinant Proteins; Serpins; Sialyl Lewis X Antigen

Primary pleomorphic adenoma of the lung is an uncommon condition. We present a case of primary pulmonary pleomorphic adenoma and its immunohistologic features. The presence of immunoreactivity to both anticytokeratin and antivimentin antibodies for its epithelial components is suggestive of a primary pulmonary lesion. Its high proliferation index and its immunoreactivity to tumor regulatory gene p16(INK4A) are features that, to our knowledge, have not been reported previously. They may have a role in the frequent recurrence of these tumors many years after their apparently complete excision. Detailed genetic investigation and long-term follow-up of this rare tumor will aid in the characterization of its biologic profile.

Topics: Adenoma, Pleomorphic; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Vimentin

Mesothelioma is a rare neoplasm of the serosal membranes. Signs and symptoms of a pleural effusion typically herald discovery of the tumor. We report a case of miliary metastatic mesothelioma involving both lungs in a 54-year-old man who presented with right-sided chest discomfort, numerous pulmonary nodules detected by computed tomography of the chest, and absent pleural effusion. Immunohistochemical and electron microscopy studies performed on wedge biopsies of parenchymal pulmonary nodules led to the diagnosis of metastatic mesothelioma. Subsequent pleural evaluation and biopsy of pleural thickening noted at a site of prior chest wall trauma identified the primary neoplasm and confirmed the diagnosis as malignant epithelioid mesothelioma. The histologic appearance of discohesive epithelioid cells in a distinctly myxoid background was the clue in this case leading to the consideration of metastatic mesothelioma and a thorough immunohistochemical evaluation of the tumor. This case shows that mesothelioma may metastasize throughout the lungs in a miliary pattern and the metastases may be clinically detected before the primary pleural tumor. Metastatic mesothelioma is a consideration for metastatic pulmonary tumors of unknown origin.

Topics: Biomarkers, Tumor; Biopsy; Calbindin 2; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Mesothelioma; Middle Aged; Neoplasms, Unknown Primary; Pleural Neoplasms; Radiography, Thoracic; S100 Calcium Binding Protein G; Tomography, X-Ray Computed

Ovarian metastasis originating from bronchioloalveolar carcinoma (BAC) has not been reported previously. We report a 63-year-old Chinese woman who was diagnosed as BAC with pleural metastasis in 1997. Four years later, she complained of vaginal bleeding, and a pelvic mass was discovered by an abdominal computerized tomography scan. Tumor debulking and total hysterectomy with bilateral salpingo-oopherectomy were performed. Pathology disclosed well-differentiated adenocarcinoma, with abundant clear cytoplasm, in the ovaries. Furthermore, immunohistochemical staining revealed that the tumor cells from the ovary and pleura were reactive to thyroid transcription factor 1 (TTF-1) and cytokeratin-7 (CK-7) but were negative for cytokeratin-20 (CK-20). The results of immunohistochemical staining, clinical course, and pathological features were compatible with the diagnosis of BAC with ovarian metastasis. In conclusion, to investigate the primary site of a metastatic ovarian cancer, clinicians should not forget the lungs since the incidence of lung cancer in females is increasing. Moreover, a monoclonal antibody panel for TTF-1, CK-7, and CK-20 may facilitate discrimination between primary and metastasized ovarian adenocarcinomas and/or identifying tumors of pulmonary origin.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Biomarkers, Tumor; Fallopian Tubes; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Middle Aged; Nuclear Proteins; Ovarian Neoplasms; Ovariectomy; Thyroid Nuclear Factor 1; Transcription Factors

Topics: Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Chorionic Gonadotropin, beta Subunit, Human; Cytotoxicity, Immunologic; Cytotoxins; Humans; Keratins; Lung Neoplasms; Neoplasm Staging; Phosphopyruvate Hydratase; Poland

We describe a case of pulmonary carcinoma with myoepithelial differentiation, analogous to basal cell adenocarcinoma of salivary glands. The patient, a 60 year-old man, smoker, presented with three peripheral nodules of the left lung. Preoperative staging was negative for metastatic disease and the patient underwent a surgical resection of the nodules. After 22 months, the patient is alive with no evidence of disease. Microscopically, the tumours were composed of atypical cells arranged in lobules, separated by basal membrane-like material. Immunohistochemically, tumour cells were positive for cytokeratin AE1/AE3, cytokeratin 14, vimentin, calponin, S-100 protein and gliofibrillary acid protein (GFAP). Electron microscopy showed features of epithelial and myoid differentiation and confirmed the myoepithelial nature of the tumour. Pulmonary tumours with myoepithelial differentiation are rare, but they have a wide and distinctive morphological spectrum.

Topics: Basement Membrane; Biomarkers, Tumor; Calcium-Binding Proteins; Calponins; Carcinoma, Basal Cell; Carcinoma, Non-Small-Cell Lung; Cell Differentiation; Epithelium; Glial Fibrillary Acidic Protein; Humans; Keratins; Lung Neoplasms; Male; Microfilament Proteins; Microscopy, Electron; Middle Aged; Myoepithelioma; Neoplasm Proteins; Organ Specificity; Remission Induction; S100 Proteins; Salivary Gland Neoplasms; Staining and Labeling; Vimentin

This report describes a benign myoepithelioma of the lung that occurred in a 60-year-old woman. The patient had experienced hoarseness for 6 weeks, and a computed tomographic scan showed a nodule of approximately 2 cm in diameter at the peripheral portion of her right upper lung. Positron emission tomography showed no uptake of F-18 fluorodeoxyglucose in the nodule. Wedge biopsy of the lesion showed benign spindle cells arranged in a whorled pattern. The cells were positive for both cytokeratin and smooth muscle actin, which corresponded to the presence of tonofilaments and myofilaments that were identified ultrastructurally. The features of the present case of benign myoepithelioma that differ from features of previously reported benign and malignant cases of myoepithelioma in the lung are discussed in the report.

Topics: Actin Cytoskeleton; Actins; Biopsy; Cell Nucleus; Cytoplasm; Deoxyglucose; Female; Fluorine Radioisotopes; Glial Fibrillary Acidic Protein; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Muscle, Smooth; Myoepithelioma; Tomography, Emission-Computed; Tomography, X-Ray Computed; Vimentin

Lung tumor markers fall into several categories including oncofetal proteins, structural proteins and their fragments, enzymes, membrane antigens, peptide and non-peptide hormones. Cytokeratins (CK) are well known structural proteins whose degradation gives rise to soluble fragments, measurable in the blood of patients and capable of cancer marking. Among them, Tissue Polypeptide Antigen (TPA), Tissue Polypeptide-Specific Antigen (TPS) and Cytokeratin-19-Fragments (Cyfra 21-1) are the most studied CK fragments' complexes. This article will review biological characteristics and clinical properties of these substances, emphasizing as their concentration in the peripheral blood might reflect the mass of tumor, the rate of cancer cell lysis, and other potentially unfavorable tumor characteristics. Assaying the concentration of CK fragments in the blood is an easy and effective way to assess lung cancer and monitor its clinical evolution.

Topics: Biomarkers, Tumor; Cytoskeleton; Disease Progression; Humans; Keratins; Lung Neoplasms; Peptides; Tissue Polypeptide Antigen

Carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), neuron-specific enolase (NSE), cytokeratin 19 fragment (CYFRA), and pro-gastrin-releasing peptide (proGRP) can be used as tumor markers for lung cancer. CEA is sensitive for adenocarcinoma, SCC and CYFRA for squamous cell carcinoma, and NSE and proGRP for small cell carcinoma. A tumor marker is generally used as a marker to monitor the clinical course. Serum levels of pro-GRP, reflect the disease course of patients with small cell lung cancer more accurately than NSE or CEA. Among the patients with clinical N0-1 non-small cell lung cancer high serum CEA levels, adenocarcinoma histology, and large tumor dimension were significant predictors of pathologic N2 disease. CEA played a new role in predicting metastasis to mediastinal lymph nodes A more effective treatment may enhance the value of tumor markers to predict relapse.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Humans; Keratins; Lung Neoplasms; Peptide Fragments; Peptides; Phosphopyruvate Hydratase; Prognosis; Recombinant Proteins; Serpins

Various evaluation methods are available for aiding clinicians in lung cancer management. Some of these methods are highly specific. However, they are also invasive and burdened by non-negligible complication rates (e.g., mediastinoscopy); other methods are highly accurate and noninvasive, but require expensive equipment and well-trained personnel (e.g., PET scanning); others are fast, inexpensive and safe. However, their diagnostic yield is low and requires further clinical testing (an example of such tests is the chest-x-ray film). There is probably only one way to perform an easy, inexpensive, repeatable test, which is also fairly accurate and predictive. This is tumor marker testing, which--as a large and specialized literature shows--can be highly effective when based on a cytokeratin-derived marker assay.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Dose-Response Relationship, Drug; Humans; Keratin-19; Keratins; Lung Neoplasms; Molecular Diagnostic Techniques; Sensitivity and Specificity

A number of serum tumor markers are clinically relevant for primary lung carcinomas. None of them, however, is applied to screening of lung cancers because of their unsatisfactory sensitivity and specificity. Among them, measurements of CEA, CYFRA 21-1, NSE, and pro GRP frequently give subsidiary information as to differential diagnosis, monitoring of treatment, and early detection of recurrence. Current trend of the tumor markers for lung cancer includes development of new markers such as p53 tumor suppressor gene product and I-CTP for detecting bone metastasis. Attempts to detect micrometastases by means of RT-PCR of these marker genes are also discussed.

Topics: Antibodies, Neoplasm; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Collagen; Collagen Type I; Humans; Keratin-19; Keratins; Lung Neoplasms; Peptide Fragments; Peptides; Recombinant Proteins; Tumor Suppressor Protein p53

In this report, we describe a highly unusual case of glandular malignant peripheral nerve sheath tumor presenting as a neck mass in a previously healthy 29-year-old man. Grossly, the tumor was found to arise from a swollen peripheral nerve trunk. The tumor was largely composed of spindle cells that demonstrated marked nuclear pleomorphism and numerous abnormal mitotic figures. In addition, histologically malignant glandular structures lined by simple nonciliated columnar cells with goblet cells were found clustered in the center of the tumor. Examination of the swollen peripheral nerve trunk revealed the presence of a plexiform neurofibroma. The spindle cells were positive for S100. The glands were negative for S100 but positive for keratin, epithelial membrane antigen, and neuroendocrine markers (somatostatin, chromogranin, Leu-7, and calcitonin). This patient was subsequently diagnosed as having von Recklinghausen disease and died of tumor metastasis to the lungs 34 months after the presentation. To our knowledge, only 3 similar cases have been previously described in the literature.

Topics: Adult; Biomarkers; Carcinoembryonic Antigen; Cell Nucleus; Epithelial Cells; Fatal Outcome; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Mitosis; Mucin-1; Nerve Sheath Neoplasms; Neurofibromatosis 1; S100 Proteins; Skin Neoplasms

Topics: Bone Marrow; Bone Marrow Examination; Bone Marrow Neoplasms; Breast Neoplasms; Colorectal Neoplasms; Esophageal Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Neoplastic Cells, Circulating; Ovarian Neoplasms; Prognosis; Prospective Studies; Randomized Controlled Trials as Topic; Risk Factors; Stomach Neoplasms; Uterine Cervical Neoplasms

Clear cell adenocarcinoma of the lung is extremely rare. On radiography, a 45-year-old female with fever was found to have an abnormal shadow in the left lower lung field. Bronchoscopy revealed a polypoid tumor in the left bronchus. On biopsy, the tumor was determined to be adenocarcinoma. Preoperative examination found no tumors outside of the lung. The patient underwent left lower lobectomy with bronchial wedge resection. The tumor had completely obstructed and dilated the left lower bronchus, but had not invaded the tissue outside the bronchial wall. Microscopically, the cytoplasm of the tumor cells contained abundant glycogen, and the tumor had solid and glandular structures. The tumor was diagnosed as clear cell adenocarcinoma of the lung.

Topics: Adenocarcinoma, Clear Cell; Bronchial Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Microscopy, Electron; Middle Aged; Mucin-1; Polyps

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Neoplasms; Neoplasms, Hormone-Dependent; Prostate-Specific Antigen; Prostatic Neoplasms; Telomerase

The increased knowledge about the molecular mechanisms leading to the development of a tumor has opened new horizons for basic and applied research. Lung cancer is among the tumor entities with the highest incidence and mortality rates. Recently new drugs and therapeutic options for patients with lung cancer were developed. These developments demand new and improved techniques for the sensitive and specific detection of lung tumor cells. Some of the molecular genetic features of lung tumor cells are summarized and the possibilities to use these characteristics as new tumor markers are discussed.

Topics: Biomarkers, Tumor; Diagnosis, Differential; Genes, ras; Humans; Keratins; Loss of Heterozygosity; Lung Neoplasms; Microsatellite Repeats; Mutation; Tumor Suppressor Protein p53

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Humans; Keratin-19; Keratins; Lung Neoplasms

Topics: Adenocarcinoma; Biomarkers, Tumor; Cadherins; Calbindin 2; Diagnosis, Differential; Humans; Hyaluronan Receptors; Immunohistochemistry; Keratins; Lung Neoplasms; Mesothelioma; Pleural Neoplasms; S100 Calcium Binding Protein G; Thrombomodulin

Neuroendocrine bladder tumours are exceptional, and the positive diagnosis is only established when they are already large and advanced. We report an original case in view of its small dimensions. We discuss the differential diagnosis (mainly bladder metastases from lung cancer) and pathological specificities, particularly the value of epithelial immunolabelling allowing exclusion of lymphoma. Because of the similarities with bronchial neuroendocrine tumours, the potential value of serum NSE assay should be emphasized. Combined surgery-cisplatin-based adjuvant chemotherapy is recommended.

Topics: Aged; Antineoplastic Agents; Carcinoembryonic Antigen; Carcinoma, Neuroendocrine; Chemotherapy, Adjuvant; Cisplatin; Diagnosis, Differential; Female; Humans; Keratins; Lung Neoplasms; Lymphoma; Mucin-1; Neoplasm Staging; Phosphopyruvate Hydratase; Synaptophysin; Urinary Bladder Neoplasms

This report describes the fine-needle aspiration (FNA) cytology of a case of pleuropulmonary blastoma in a 3-yr-9-mo-old male. Pleuropulmonary blastoma is considered by most authors to be distinct from pulmonary blastoma and is a rare malignant tumor of the intrathoracic cavity. FNA smears were cellular with numerous small ovoid to spindled cells with oval to elliptical nuclei exhibiting finely granular chromatin and inconspicuous nucleoli. The cytoplasm was scant and eosinophilic with indistinct borders. Focal chondroid material and blastema-like cells were noted. The differential diagnosis suggested by the cytologic findings included rhabdomysosarcoma, teratoma, neuroblastoma, malignant mesenchymoma, pleuropulmonary blastoma, and metastatic tumor. To our knowledge, this is the first report of the cytology of this tumor.

Topics: Actins; Biopsy, Needle; Child, Preschool; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Pleural Neoplasms; Pulmonary Blastoma; S100 Proteins; Vimentin

Sarcomatoid carcinomas (SC) of the lung are the most common pulmonary neoplasms that exhibit a composition by spindled or pleomorphic tumor cells. As such, many of them may be confused easily with true sarcomas diagnostically unless special immunohistological or ultrastructural analyses are performed. Reactivity is expected for keratin, epithelial membrane antigen, or collagen type IV in the sarcomalike elements in SC, although it may be focal. Electron microscopy often shows the presence of junctional complexes between tumor cells, with or without pericellular basal lamina and cytoplasmic skeins of intermediate filaments. Current terminological preferences are such that several formerly used terms--including "spindle-cell carcinoma," "pulmonary blastoma," "squamous cell carcinoma with pseudosarcomatous stroma," "pseudosarcoma," and "carcinosarcoma"--are now encompassed by the more generic designation of "sarcomatoid carcinoma." The clinical course of patients with this neoplasm is aggressive, with an overall 5-year survival rate approximating 20%.

Topics: Carcinosarcoma; Collagen; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mucin-1; Pulmonary Blastoma

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Humans; Keratin-19; Keratins; Lung Neoplasms; Mass Screening; Neoplasm Staging; Phosphopyruvate Hydratase; Prognosis; Sensitivity and Specificity

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Humans; Keratin-19; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase; Prognosis; Survival Analysis; Treatment Outcome

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Drug Monitoring; Humans; Keratin-19; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase; Treatment Outcome

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Humans; Keratin-19; Keratins; Lung Neoplasms; Neoplasm Recurrence, Local; Phosphopyruvate Hydratase; Sensitivity and Specificity

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Bronchogenic; Humans; Keratin-19; Keratins; Lung Neoplasms; Prognosis; Reproducibility of Results; Sensitivity and Specificity; Treatment Outcome

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Humans; Keratin-19; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase; Sensitivity and Specificity

Sclerosing mucoepidermoid carcinoma with eosinophilia (SMECE) is a recently recognized malignant neoplasm of the thyroid gland. Two additional cases of this condition which occurred in a 70-year-old woman and a 69-year-old woman are presented. The case of the 70-year-old woman (patient 1) is the first report of distant metastasis, besides lymph node metastasis, for this type of tumor. The patient initially presented with a thyroid mass, and the thyroid gland with surrounding cervical lymph nodes was removed. Because of focal keratin "pearl" formation, the tumor was misinterpreted as a metastatic squamous cell carcinoma to the thyroid. Approximately 4 years later, the patient developed a left supraclavicular mass and lung densities. A pathological fracture of the right humeral head followed, and the left supraclavicular mass recurred along with newly developed subcutaneous nodules on the chest wall and arm. Open lung and bone biopsies revealed metastatic SMECE, which was morphologically identical to that of the thyroid mass. The 69-year-old woman (patient 2) had, in 1983, undergone thyroidectomy with left radical neck dissection; this had been diagnosed as follicular carcinoma of the thyroid with lymph node involvement. After multiple isolated lymph nodes metastases, the patient developed locally extensive, recurrent tumor that showed microscopic features of SMECE. Review of the previous thyroid tumor and lymph nodes revealed the same type of histology. To our knowledge, only a single report containing eight cases of this distinctive carcinoma of the thyroid has been published. Herein we describe characteristic morphological features of two additional cases of this rare malignancy, one with distant metastasis, and we review the related literature.

Topics: Aged; Carcinoma, Mucoepidermoid; Carcinoma, Squamous Cell; Diagnosis, Differential; Eosinophilia; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Sclerosis; Thyroid Neoplasms; Thyroiditis, Autoimmune

The problems with classification and diagnosis of bronchopulmonary carcinoid and other neuroendocrine tumors are described in this paper. Single neuroendocrine cells and so-called neuroepithelial bodies found in normal bronchial epithelium are currently believed to constitute pulmonary components of an extensive neuroendocrine system. In view these opinions, it has appeared a need for a new, standardized nomenclature. Hence presently suggested classification of neuroendocrine carcinomas taking into account their histological structure, immunohistochemical as well as prognostic features starting from carcinoid tumors to small cell anaplastic carcinomas-includes three types of neoplasms.

Topics: Biomarkers, Tumor; Bronchial Neoplasms; Carcinoid Tumor; Carcinoma, Neuroendocrine; CD57 Antigens; Cell Size; Chromogranin A; Chromogranins; Humans; Keratins; Lung Neoplasms; Neoplasm Proteins; Neuroendocrine Tumors; Neuropeptides; Phosphopyruvate Hydratase; Silver Staining; Somatostatin; Synaptophysin; Terminology as Topic

In Okinawa, a subtropical island in southern Japan, squamous cell carcinoma (SCC), especially the well-differentiated form, is prevalent, while this form is relatively rare in both the mainland and other countries (e.g. United States of America). More patients with SCC from Okinawa, moreover, were positive for human papillomavirus (HPV) DNA by polymerase chain reaction (PCR) (79%), and harbored HPV types 6, 16 and 18, in combination. On the other hand, less than 30% of the mainland patients were positive for HPV DNA by PCR. Those patients who were positive all harbored only one HPV type. Furthermore, in Okinawa, there were a significant number of cases with adenosquamous carcinoma, and they too were positive for HPV DNA. The SCC and the adenocarcinoma cells adjacent to the SCC component in these cases were also positive for HPV DNA, and such adenocarcinoma cells were enlarged in size with relatively wide cytoplasm. The authors postulate that HPV infects adenocarcinoma cells and changes them to enlarged cells, followed by squamous metaplasia. In this report, HPV DNA was transfected to adenocarcinoma cells (cultured cell lines) and this showed that HPV causes squamous metaplasia. In addition, aberrant expression of p53 was demonstrated in a large number of the SCC cases in Okinawa. The enlarged adenocarcinoma cells adjacent to the SCC components in adenosquamous carcinomas also showed aberrant expression of p53. The recent advances in the studies of anti-oncogenes, p53, etc. and oncogenes are outlined. It is to be noted that the molecular mechanisms of carcinogenesis in the lung have been studied in general, classifying lung tumors into two groups, namely, small cell carcinoma (SCLC) and non-small cell carcinoma (NSCLC). However, because human lung cancer is represented by a wide variety of histologic types, molecular genetic studies according to a more detailed histological subclassification is needed.

Topics: Adult; Aged; Aged, 80 and over; Animals; Blotting, Southern; Blotting, Western; Carcinoma, Adenosquamous; Carcinoma, Squamous Cell; Female; Humans; In Situ Hybridization; Japan; Keratins; Lung Neoplasms; Male; Metaplasia; Mice; Mice, SCID; Middle Aged; Mutation; Papillomaviridae; Transfection; Tumor Cells, Cultured; Tumor Suppressor Protein p53

Topics: Actin Cytoskeleton; Aged; Basement Membrane; Carcinoma, Adenoid Cystic; Cytoplasmic Granules; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Microvilli; Mitochondria; S100 Proteins; Salivary Gland Neoplasms; Salivary Glands, Minor; Solitary Pulmonary Nodule; Tongue Neoplasms

Topics: Biomarkers, Tumor; Humans; Keratins; Lung Neoplasms; Peptide Fragments

Malignant mesenchymomas are uncommon tumors of soft tissues. Three such tumors involving the pleura have been reported in the literature. We report a case of malignant mesenchymoma of the pleura that had liposarcomatous, rhabdomyosarcomatous, chondrosarcomatous, and osteosarcomatous elements.

Topics: Adipose Tissue; Aged; Carcinoma, Large Cell; Diagnosis, Differential; Humans; Keratins; Lung Neoplasms; Male; Mesenchymoma; Necrosis; Pleural Neoplasms

In the clinical practice of lung cancer, serum tumor markers are important laboratory tests and their use is wide spread. Among the various markers for lung cancer, the usefulness of CEA, SLX, SCC and NSE has been firmly established. These markers cannot be used routinely to screen for lung cancer but may be used as complementary tools for diagnosing the tumor. Elevated levels of these markers also appear to be useful for monitoring the response to therapy and tumor progression. CYFRA21-1 and ProGRP, new tumor markers with relatively high sensitivity and specificity to lung cancer, were recently developed. These tumor markers may be useful tools for early detection of lung cancer.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Diagnosis, Differential; Gastrointestinal Hormones; Humans; Intermediate Filament Proteins; Keratin-19; Keratins; Lewis X Antigen; Lung Neoplasms; Neoplasm Staging; Peptide Fragments; Peptides; Phosphopyruvate Hydratase; Recombinant Proteins

Topics: Animals; Carcinoma, Non-Small-Cell Lung; Disease Models, Animal; Genetic Therapy; Humans; Keratins; Lung Neoplasms; Neoplasm Transplantation; Transplantation, Heterologous

Within the past few years, the measurement of serum and tissue markers has had an increasing influence on clinical decisions about initial treatment and follow-up. Lung cancer illustrates the types and importance of these various markers. This review presents data concerning the most studied and interesting markers in non-small cell (NSCLC) and small cell lung cancer (SCLC). CEA, TPA, SCC-Ag, CYFRA 21-1, ferritin, CA19-9, CA50, CA242, H-K-N-ras mutations and p53 mutation seem to be the most prolific in NSCLC, while NSE, BN/GRP, CK-BB, NCAM, IL-2R, IGF-I, transferrin, ANP, mAb (cluster 5), Le-y and c-N-L-myc mutation are markers in SCLC patients. Some of these serum markers might be useful adjuncts for monitoring response to therapy, including early detection of tumour reactivation to allow curative therapy and rapid detection of treatment failure to allow change of the regimen. The study of these markers also may lead to a better understanding of the biological characteristics of lung cancer. The information derived from these biological studies represents the most promising avenue towards new treatment strategies, as well as attempts at secondary prevention.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Genes, p53; Genes, ras; Humans; Keratins; Lung Neoplasms; Peptides; Phosphopyruvate Hydratase; Tissue Polypeptide Antigen

CYFRA 21-1 has proved to be a useful marker for non-small-cell lung cancer (NSCLC), which is the major form of lung cancer. Its most effective application is in monitoring. CYFRA 21-1 provides diagnostic information about the success of primary surgery, the response to chemotherapy and the detection of relapse. It is also an independent prognostic factor. The diagnostic potential may not be fully used because decision-making is currently based on group reference ranges. It seems useful to carry out systematic studies to investigate if the application can be improved and extended by using individual reference ranges for decision-making. In contrast to most of the other tumour markers the comparability of the commercial CYFRA 21-1- assays currently available on the market is good. The high degree of comparability should be maintained by international standardization. The analytical performance of the currently available commercial CYFRA 21-1 tests meets requirements derived from its current clinical applications. However, there are no data available about the analytical performance under field conditions. CYFRA 21-1, an established tumour marker for lung cancer, should be included in external quality assurance schemes.

Topics: Biomarkers, Tumor; Humans; Keratins; Lung Neoplasms; Predictive Value of Tests

This report is a case of epithelioid hemangioendothelioma presenting as multiple lytic lesions of the ilium with radiographic findings of diffuse, bilateral lung involvement and biopsy-proven scalp involvement. Histologically, the tumor within bone and skin exhibited cords and nests of plump, epithelioid-appearing cells exhibiting rudimentary vascular differentiation within a myxohyaline stroma. Aggressive histologic features were not present. Immunohistochemical reactivity for Factor VIII-related antigen, Q-bend 10 (CD34), and cytokeratin were demonstrated. Ultrastructural studies revealed abundant intermediate cytoplasmic filaments, pinocytotic vacuoles, and Weibel-Palade bodies. The concurrent bone, skin, and lung involvement, low-grade histologic type, and female sex of the patient aroused speculation about the role of hormones in the development and possible treatment of the tumor, but estrogen and progesterone receptors were not detected. Despite intense combination chemotherapy, the patient died of widely metastatic disease. This report demonstrates the aggressive potential of histologically low-grade epithelioid hemangioendothelioma and the need for a thorough evaluation for metastases.

Topics: Adult; Antigens, CD; Antigens, CD34; Female; Hemangioendothelioma, Epithelioid; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Neoplasms, Second Primary; Receptors, Estrogen; Receptors, Progesterone; Skin Neoplasms; Tomography, X-Ray Computed; von Willebrand Factor

Topics: Adult; Digestive System Neoplasms; Endocrine Gland Neoplasms; Epithelium; Female; Humans; Keratins; Lung Neoplasms; Male; Neoplasms; Precancerous Conditions; Skin Neoplasms; Urogenital Neoplasms

Approximately 700 cases of keratinizing cystic squamous lung lesions in rats were investigated by light microscopy in order to clarify the nomenclature and classification of these lesions. The structure of benign keratinizing cystic squamous cell tumours of the lung was compared to that of cystic squamous lesions in the skin of rats, with consideration of data from the literature. We conclude that the reviewed keratinizing cystic squamous cell lesions of the lung are true neoplasms and that the growth pattern of these cystic lesions is inconsistent with that of a simple cyst. In the development of squamous lung cancer, a continuum of proliferation from exaggerated metaplasia through benign cystic tumours to invasive squamous cell carcinomas can be observed.

Topics: Animals; Epidermal Cyst; Keratins; Keratoacanthoma; Lung Neoplasms; Rats; Skin Diseases; Terminology as Topic

We evaluated three cases of pigmented pulmonary carcinoid tumors that were retrieved from the files of the Armed Forces Institute of Pathology, Washington, DC. Clinical follow-up showed no indication of tumor recurrence or metastases, nor was there evidence of malignant melanoma. All three cases exhibited histologic features of typical carcinoid tumor; there were focal oncocytic changes in two cases. Finely dispersed, brown pigment, believed to be melanin, was distributed in two different patterns: in sustentacular cells (case 1) or within the tumor cells (cases 2 and 3). Fontana-Masson stain was positive in areas of this pigment in all cases. The tumor cells showed immunoreactivity for chromogranin, synaptophysin, keratin (AE1/AE3 and CAM-5.2), and S100 protein in all cases. Focal staining for vimentin and corticotropin was seen within neoplastic cells in two cases. The pigmented sustentacular cells in case 1 showed focal immunoreactivity for S100 protein and HMB-45. Ultrastructural studies of paraffin-embedded tissues were performed in two cases. They showed well-developed melanosomes in the pigmented sustentacular cells in case 1. In both cases, cytoplasmic neurosecretory-type granules were identified in neoplastic cells. These findings demonstrate that pigmented pulmonary carcinoid tumor has an immunohistochemical profile similar to that of typical pulmonary carcinoid tumor. In some instances, pigmented pulmonary carcinoid tumors may show ultrastructural evidence of melanocytic and neuroendocrine differentiation. These immunohistologic and ultrastructural findings distinguish pigmented pulmonary carcinoid tumor from malignant melanoma and support the concept of "multidirectional cellular differentiation."

Topics: Adult; Aged; Antibodies, Monoclonal; Carcinoid Tumor; Chromogranins; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Pigmentation; S100 Proteins; Synaptophysin

Merkel cell carcinomas (MCC) were compared to small cell carcinomas of the lung (SCCL) and oesophagus (SCCO). Most MCC were of the intermediate cell type while SCCL and SCCO were usually of the small cell type. Only MCC of trabecular type could be separated from SCCL and SCCO by means of histopathological examination alone. All MCC (25) stained with cytokeratin CAM 5.2, 20 of which in a "paranuclear globular" or combined "paranuclear globular"/diffuse pattern while 17 MCC stained with cytokeratin AE1/AE3. Cytokeratin CAM 5.2 reacted with 60 percent of the SCCL and 86 percent of the SCCO, and cytokeratin AE1/AE3 with 33 and 28 percent respectively. Neurofilament stained 17 MCC in a "paranuclear globular" pattern but none of the SCCL and SCCO. All MCC with a diffuse staining pattern for cytokeratin CAM 5.2 were negative for neurofilament. The results of this study and review of the literature indicate that in most instances Merkel cell carcinoma can be separated from other SCC, pulmonary as well as extrapulmonary, by means of histopathological and, above all, immunohistochemical examinations.

Topics: Antigens, CD; Antigens, Differentiation; Carcinoma, Merkel Cell; Carcinoma, Small Cell; Esophageal Neoplasms; Histocompatibility Antigens; Humans; Intermediate Filament Proteins; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Membrane Glycoproteins; Mucin-1; Neurofilament Proteins; Phosphopyruvate Hydratase; S100 Proteins

The fifteenth case of malignant Sertoli cell tumour is reported and the literature is reviewed. The reported case was unilateral with lung metastases. Immunohistochemical examination showed positive staining reaction within the tumour cells for vimentin and cytokeratin, while AFP, HCG, PLAP, EMA and CEA were not found, which is in accordance with the staining pattern found in normal Sertoli cells.

Topics: Aged; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Sertoli Cell Tumor; Testicular Neoplasms; Vimentin

Topics: Adenocarcinoma; Adenoma, Islet Cell; Animals; Antibodies, Monoclonal; Breast Neoplasms; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cytoskeletal Proteins; Desmoplakins; Desmosomes; Electrophoresis, Polyacrylamide Gel; Granulosa Cell Tumor; Humans; Immunosorbent Techniques; Intermediate Filament Proteins; Intermediate Filaments; Keratins; Kidney Neoplasms; Lung Neoplasms; Melanoma; Membrane Proteins; Meningioma; Mesothelioma; Microscopy, Electron; Microscopy, Fluorescence; Neoplasms; Neurosecretory Systems; Skin Neoplasms

This diagnostic seminar discusses the current status of the principles and problems of cytology as it is applied to the diagnosis of lung cancer. This discussion is divided into four major parts. Part I presents a discussion of cytopreparatory techniques and cytology of the lung in the absence of cancer. The cytology of benign proliferations which may mimic cancer is emphasized. The role of cytology in the diagnosis of pulmonary infectious organisms is noted. Part II discusses lung cancer as manifested in specimens of sputum, bronchial washings, and bronchial brushings. Part III presents some data on the validity of cytology with respect to role of specimen number and type in lung cancer diagnosis and cell typing in lung cancer. The continued usefulness and importance of multiple specimens of sputum for lung cancer diagnosis are documented. Part IV presents a brief synopsis of fine needle aspiration biopsy of lung cancer.

Topics: Adenocarcinoma; Aspergillus; Biopsy, Needle; Blastomyces; Bronchi; Carcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Nucleus; Coccidioides; Cryptococcus neoformans; Cytodiagnosis; Cytological Techniques; Cytoplasm; Epithelium; Histoplasma; Humans; Keratins; Lung Diseases; Lung Diseases, Fungal; Lung Diseases, Parasitic; Lung Neoplasms; Macrophages; Metaplasia; Pneumocystis; Sputum; Strongyloides; Suction; Virus Diseases

Topics: Antibodies, Monoclonal; Carcinoma, Small Cell; Cell Line; Desmosomes; Humans; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Neoplasms; Neuroectodermal Tumors, Primitive, Peripheral; Rhabdomyosarcoma

A wide variety of human neoplasms were examined by immunocytochemical and ultrastructural techniques. In most, one intermediate filament (IF) type was expressed reflecting the tissue of origin. However, multiple classes of intermediate filaments were regularly found in a subgroup of these tumors. We chose to subdivide them into those with a complex or mixed growth pattern, and those which showed a more "monomorphic" histologic growth pattern. This latter group is the subject of this paper. Regular coexpression of cytokeratin and vimentin was observed in tumors of endometrial, thyroid, ovarian and renal origin, and coexpression of cytokeratin and neurofilament was observed in a subgroup of neuroendocrine tumors. Immunocytochemical/ultrastructural correlation demonstrated few, if any, observable intermediate filaments in tumors expressing only low molecular weight cytokeratin, whereas vimentin and neural filament characteristically were randomly dispersed or formed whorled bundles of cytoplasmic filaments. The potential diagnostic usefulness of these observations in surgical pathology is discussed.

Topics: Carcinoid Tumor; Carcinoma; Carcinoma, Small Cell; Cytoskeleton; Endocrine System Diseases; Female; Humans; Intermediate Filaments; Keratins; Kidney Neoplasms; Lung Neoplasms; Ovarian Neoplasms; Thyroid Neoplasms; Uterine Neoplasms; Vimentin

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Humans; Keratin-19; Keratins; Lung Neoplasms; Mesothelioma; Mesothelioma, Malignant; Prognosis

Currently, the diagnostic sensitivity of malignant liver mass biopsies is an important problem in the definitive diagnosis. In this study, we aimed to investigate the role of selective peripheral approach to lesion biopsies for diagnostic sensitivity of liver masses.. Between June 2007 and March 2011, totally 88 patients (50 male, 38 female), referred to our Interventional Radiology Department for sonographically guided Tru-cut biopsies for liver lesions, were examined.All biopsies were performed by an experienced radiologist with an 18-gauge Tru-cut biopsy needle with a spring-loaded biopsy gun under sonographic guidance. We describe two locations (peripheral and central) for liver lesions, with the inner 2/3 part of the mass as central and the outer 1/3 part as peripheral. We obtained biopsy from both of these locations, and samples were transferred to the Pathology Department separately.. According to pathological and immunohistochemistry studies, there were 42 hepatocellular carcinomas and 46 metastases. All of the metastatic tumors were stained by cytokeratin (10 lung adenocarcinoma, 15 breast adenocarcinoma, 16 gastrointestinal tract, 4 prostate, and 1 malignant melanoma of these 46 metastases were reported as primary). According to histopathological results, diagnostic sensitivity was 97.7% in peripherally located biopsies and 86.3% in biopsies taken from the center of the masses (p=0.0063).. Selective peripheral biopsy approach in Tru-cut biopsies of liver lesions has better sensitivity rates for histopathologic diagnosis compared to the centrally located and random biopsies.

Topics: Adenocarcinoma; Adult; Aged; Biomarkers, Tumor; Biopsy, Needle; Breast Neoplasms; Carcinoma, Hepatocellular; Creatine Kinase; Diagnosis, Differential; Female; Gastrointestinal Neoplasms; Humans; Keratins; Liver Cirrhosis; Liver Neoplasms; Lung Neoplasms; Male; Melanoma; Middle Aged; Prostatic Neoplasms; Sensitivity and Specificity; Skin Neoplasms

Circulating tumor cell (CTC) counts are an established prognostic marker in metastatic prostate, breast and colorectal cancer, and recent data suggest a similar role in late stage non-small cell lung cancer (NSCLC). However, due to sensitivity constraints in current enrichment-based CTC detection technologies, there are few published data about CTC prevalence rates and morphologic heterogeneity in early-stage NSCLC, or the correlation of CTCs with disease progression and their usability for clinical staging. We investigated CTC counts, morphology and aggregation in early stage, locally advanced and metastatic NSCLC patients by using a fluid-phase biopsy approach that identifies CTCs without relying on surface-receptor-based enrichment and presents them in sufficiently high definition (HD) to satisfy diagnostic pathology image quality requirements. HD-CTCs were analyzed in blood samples from 78 chemotherapy-naïve NSCLC patients. 73% of the total population had a positive HD-CTC count (>0 CTC in 1 mL of blood) with a median of 4.4 HD-CTCs mL⁻¹ (range 0-515.6) and a mean of 44.7 (±95.2) HD-CTCs mL⁻¹. No significant difference in the medians of HD-CTC counts was detected between stage IV (n = 31, range 0-178.2), stage III (n = 34, range 0-515.6) and stages I/II (n = 13, range 0-442.3). Furthermore, HD-CTCs exhibited a uniformity in terms of molecular and physical characteristics such as fluorescent cytokeratin intensity, nuclear size, frequency of apoptosis and aggregate formation across the spectrum of staging. Our results demonstrate that despite stringent morphologic inclusion criteria for the definition of HD-CTCs, the HD-CTC assay shows high sensitivity in the detection and characterization of both early- and late-stage lung cancer CTCs. Extensive studies are warranted to investigate the prognostic value of CTC profiling in early-stage lung cancer. This finding has implications for the design of extensive studies examining screening, therapy and surveillance in lung cancer patients.

Topics: Adult; Aged; Aged, 80 and over; Biopsy; Carcinoma, Non-Small-Cell Lung; Disease Progression; Female; Humans; Image Interpretation, Computer-Assisted; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplastic Cells, Circulating; Prognosis

Sentinel lymph node identification has been tested in lung cancer patients with conflicting results. The present study was designed to assess the sensitivity, negative predictive value, and accuracy of intraoperative sentinel lymph node mapping by means of a radio-guided method in patients with nonsmall cell lung cancer to find the most appropriate definition of sentinel lymph node and to evaluate the usefulness of different particle sizes of radiocolloid.. One hundred ten patients with clinically N0 nonsmall cell lung cancer were enrolled in the pilot study of intraoperative sentinel node identification. Four quadrants of the peritumoral tissue were injected with 2 mL of 0.5 mCi technetium-99m suspension. Four radiocolloids of different particle size were used. After complete lymphadenectomy, all resected lymph nodes were examined with hematoxylin-eosin staining. All sentinel nodes negative for metastases by routine staining were searched further for metastatic deposits with both serial sections and immunohistochemistry for cytokeratins.. The radio-guided method had a high identification rate, a high sensitivity, and a high negative predictive value (100%, 87%, and 93%, respectively) when immunohistochemistry was considered. When standard hematoxylin and eosin staining was applied, sensitivity and negative predictive value of sentinel lymph node labeling was lower (74% and 89%, respectively). No significant differences were found in either the sensitivity or negative predictive value among the colloid solutions of different particle size used in radio labeling, although smaller particles have shown a tendency to produce better results.. The radio-guided technique provides efficient sentinel lymph node identification in lung cancer. Further studies are warranted to confirm the clinical utility of this strategy.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Decision Making; Female; Humans; Intraoperative Care; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Mediastinum; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Particle Size; Pilot Projects; Pneumonectomy; Predictive Value of Tests; Radionuclide Imaging; Radiopharmaceuticals; Reproducibility of Results; Sensitivity and Specificity; Sentinel Lymph Node Biopsy; Technetium Tc 99m Aggregated Albumin; Technetium Tc 99m Sulfur Colloid

To investigate the changes and clinical value of circulating endothelial cells (CEC) in the peripheral blood of advanced NSCLC patient.. Sixty-seven advanced NSCLC patients were randomly divided into either the treatment group with NP plus endostatin or control group with NP alone. Level of CEC and cytokeratin (CK) in the peripheral blood were measured by flow cytometry.. The response rate and benefit rate was 44.4%, 80.0% in the treatment group, and 27.3%, 50.0% in the control group, respectively (P = 0.176 and P = 0.012). Time to tumor progression (TTP) was 146.7 days in the treatment group and 91.1 days in the control group (P = 0.061). However, when the cut-off of TTP was defined as > 170 days, there was a significant difference between two groups (cut-off = 170, P = 0.034; cut-off = 180, P = 0.009). The number of CEC decreased by 0.29 +/- 0.47 in the treatment group and by 0.01 +/- 0.43 in the control group (P = 0.033). The correlation between CEC and CK was found to be positive either before (r = 0.381, P = 0.013) or after the treatment (r = 0.450, P = 0.004).. Chemotherapy combined with endostatin is superior to chemotherapy alone in the treatment of NSCLC. CEC, as a biomarker, may be useful in predicting the efficacy of the combined treatment.

Topics: Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Non-Small-Cell Lung; Cell Count; Cisplatin; Endostatins; Endothelial Cells; Endothelium, Vascular; Female; Flow Cytometry; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplastic Cells, Circulating; Remission Induction; Treatment Outcome; Vinblastine; Vinorelbine

Metastatic lymph nodes (LNs) are the major prognostic factor in resected non small cell lung carcinoma (NSCLC). However, almost 50% of pN0 patients relapse, suggesting metastatic cells undetected by current staging procedures. A combination of markers [cytokeratins 19 and 7 (CK19, CK7) and mucin type 1 (MUC1) mRNAs] was therefore evaluated by real-time RT-PCR in order to detect occult cancer cells. Forty-three NSCLC tumor samples, 4 micrometastatic, 6 metastatic and 84 histologically negative mediastinal LNs from 19 patients with NSCLC were evaluated as well as blood mononuclear cells from 29 healthy volunteers and 17 benign LNs. When tested on cell lines, RT-PCR was particularly efficient for evaluation of CK19, CK7 and MUC1 mRNA expression. All tumor samples were positive for at least 1 marker and 74% of samples were positive for all 3 markers. CK7 and CK19 mRNA were not detected in benign LN and blood cells from healthy donors in contrast with MUC1 mRNA. Only CK7 and CK19 mRNA were therefore used for evaluation of mediastinal LNs: the 6 histologically metastatic and the 4 micrometastatic LNs were positive for at least one marker. Among the 84 histologically negative LNs, 6 (7%) were positive for at least one marker, potentially changing the stage of 2 out of 19 patients. In conclusion, in our feasibility study, parallel molecular detection of CK19 and CK7 mRNA can be considered a specific diagnostic tool for the assessment of microscopic lymphatic spread. Its prognostic impact remains to be evaluated in a prospective study.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratin-7; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Mucin-1; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Luminescent Measurements; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Reference Values; Sensitivity and Specificity

The detection of isolated tumor cells (ITC) in the bone marrow of patients with epithelial malignancies is an independant prognostic factor for several entities as breast cancer, colorectal cancer or non-small lung cancer. However, with conventional immunocytology using Ficoll density gradient and APAAP staining, only a small proportion of the bone marrow samples can be scanned for cytokeratin-positive (CK+) cells. To improve detection rates, we evaluated the enrichment of ITC by magnetic activated cell sorting (MACS) compared to regularly stained cytospins. Recovery experiments with a CK+ breast cancer cell line (SKBR3) were performed to calculate the MACS enrichment rate. Bone marrow was obtained by aspiration from 20 patients with carcinomas of epithelial origin and from 17 controls. ITC were enriched and stained with magnetically labeled CAM 5.2 antibodies directed to cytokeratin 7 and 8. MACS of SKBR3 seeded in peripheral blood revealed average recovery rates of 62% and 48% and average enrichment factors of 104-fold and 8139-fold of the CK+ cells after one and after two separations, respectively. After immunomagnetic enrichment, CK+ cells were detected in 16 of 20 (80%) cancer patients, whereas only 7 (35%) patients showed CK+ cells without magnetic enrichment (P = 0.002). Ten of twelve (83%) patients with metastatic disease (stage M1) and six of eight (75%) patients without any overt metastases (M0) had CK+ cells in their bone marrow. None of the negative controls showed any CK+ cells. Enrichment with magnetically labeled anti cytokeratin antibodies increases the detection rate of epithelial cells in bone marrow of cancer patients compared to immunocytology.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Biomarkers, Tumor; Bone Marrow Neoplasms; Breast Neoplasms; Carcinoma; Carcinoma, Non-Small-Cell Lung; Colonic Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Magnetics; Male; Middle Aged; Prospective Studies; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Cyfra 21-1 is a tumor marker based on the determination of water-soluble cytokeratin 19 which is secreted by normal or malignantly transformed epithelial cells. It is suggested to be a valuable marker in patients with non-small cell lung carcinoma (NSCLC). A prospective clinical study was conducted to investigate the value of Cyfra 21-1 for diagnosis, determination of subtypes, staging, and evaluation of therapy response in patients with lung carcinoma (Ca). Sixty-nine patients (mean age: 60.9 +/- 9.2 years, M/F:12.8) treated between 1994 and 1998 inclusive, and 13 healthy smokers (mean age:50.9 +/- 4.8 years, M/F:1.6) constituted the study group and control group, respectively. Venous blood samples (10 ml) were obtained from all subjects. Posttreatment blood samples were also obtained from 14 NSCLC patients. Cyfra 21-1 levels (cutoff value 3.3 ng/ml) were determined by ELSA-Cyfra 21-1 kit (CIS bio international, France) through immunoradiometric assay (IRMA). Cyfra 21-1 levels did not differ between smoking and non-smoking subjects within each group (p > 0.05). Cyfra 21-1 was significantly elevated in lung Ca cases irrespective of the cell type (p < 0.05). It was significantly elevated in squamous cell and adenocarcinoma varieties with the most prominent elevation in squamous cell type (p < 0.05). In lung Ca, the specificity and sensitivity of Cyfra 21-1 was 92.3% and 52.2%, respectively. Sensitivity was 65.5% for NSCLC, 70.5% for squamous cell, and 45.5% for adenocarcinoma varieties, with highest sensitivity rates in Stage IIIA + IIIB (87.5%) and Stage IV (75%) of squamous cell lung Ca. Cyfra 21-1 level was significantly decreased after treatment in NSCLC patients (n 14) (p < 0.01). Cyfra 21-1 is a tumor marker that helps to establish the diagnosis and differentiation of cell type and evaluation of response to therapy in patients with NSCLC.

Topics: Analysis of Variance; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Prospective Studies

Serum cytokeratin 19 fragment (CYFRA 21-1) and carcinoembryonic antigen (CEA) levels were determined with an enzyme immunoassay in 51 patients with non-small cell lung cancer (NSCLC), 26 patients with benign lung diseases and 26 normal individuals in order to evaluate their clinical utility in the diagnosis of NSCLC. Patients with NSCLC demonstrated higher serum CYFRA 21-1 and CEA levels than both patients with benign lung diseases and normal group. We used the cut off value which was derived from the 95th percentile value of CYFRA 21-1 and CEA levels in the group of patients with benign lung diseases; CYFRA 21-1 at 3.13 ng/ml and CEA at 7.7 ng/ml. The sensitivity and diagnostic accuracy of CYFRA 21-1 and CEA for the group of NSCLC patients were 66.7 per cent, 76.6 per cent and 35.3 per cent, 55.8 per cent, respectively. When combining CYFRA 21-1 with CEA, the sensitivity and diagnostic accuracy were 68.6 per cent and 66 per cent. These results suggest that CYFRA 21-1 and CEA are useful serum markers for the diagnosis of NSCLC; especially subtype squamous cell and adenocarcinoma, respectively. The usefulness is not enhanced by combining the assay of CYFRA 21-1 and CEA.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Probability; Reference Values; Sensitivity and Specificity

To study the expression of keratin 19, 20 (K19, K20) in different tumor cell lines and tumor tissues and its clinical implication.. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was employed to examine the specific expression of K19 and K20 mRNA in eleven tumor cell lines and 33 corresponding tumor tissue specimens.. The expression of K19 mRNA was detected in 4 kinds of tumor cell lines and all tumor tissues examined, but the magnitude of expression differed, with a difference ranging from 1.7 to 10 folds for the same type of cancer. In some patients, the level of expression was as low as 12% of the positive control. K20 mRNA expression was negative for lung and esophageal tumor cell lines and the corresponding carcinoma specimens. In one of 6 bladder cancer specimens and in 4 of 5 colorectal cancer tissues, K20 expression was positive, at a level of 41%-77% of the positive control. There was no expression of K20 in bladder tumor cell line EJ1 and colorectal tumor cell line SW480.. These results demonstrate that K19 and K20 may be used as a valuable marker for detecting circulating cancer cells, but the low level of expression in some cases of carcinoma would probably result in false negative results.

Topics: Biomarkers, Tumor; Colorectal Neoplasms; Down-Regulation; Esophageal Neoplasms; Humans; Intermediate Filament Proteins; Keratin-20; Keratins; Lung Neoplasms; Neoplasms; Neoplastic Cells, Circulating; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Lung cancer, of which non-small-cell lung cancer (NSCLC) constitutes about 80%, is the greatest cause of cancer-related death worldwide. Serum tumour markers may be helpful in early diagnosis, in the initial assessment of the progress of the disease and in monitoring of the tumour growth or tumour volume reduction. Recent studies have focused on a new family of markers--hematopoietic growth factors. Some clinical investigations have shown autologous production of stem cell factor (SCF) in various human cell lines derived from lung cancer and the expression of SCF mRNA in these lines. In this study, the serum level of SCF was measured using a sensitive sandwich ELISA system in 34 patients with non-small-cell lung cancer before and 10, 30, 90, 180 and 270 days after operation. Additionally common accepted tumour markers such as CEA and CYFRA 21.1 were also assayed. Preoperative level of SCF was increased in cancer patients in comparison to the normal sera. Concentrations of SCF and CYFRA 21.1 were decreased on 10th day, but CEA on 30th day after surgical treatment, although upon comparison of pre- and postoperative tumour markers serum levels significant difference was observed for SCF and CYFRA 21.1 (p < 0.05). Levels of SCF were increased in 79%, CEA in 62% and CYFRA 21.1 in 51%. The diagnostic sensitivity of SCF were related to the stage of the disease and the combined use of two markers increased the sensitivity compared with the use of only one. These results suggest that SCF may be useful in the diagnostic and monitoring of patients with NSCLC.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Sensitivity and Specificity; Stem Cell Factor

Several circulating tumour markers for non-small-cell lung cancer (NSCLC) have been identified. Recent studies have focused on a new family of markers--hematopoietic growth factors. Some clinical investigations have shown cell surface receptors for interleukin 3 (IL-3) in lung cancer cells and autologous production of IL-3 in various human cell lines derived from NSCLC. The purpose of this investigation was to compare serum levels of IL-3 in non-small-cell lung cancer with a control group, to assess pre- and post treatment levels of IL-3 in relation to levels of commonly accepted tumour markers such as carcino-embryonic antigen (CEA) and cytokeratin fragment 19 (CYFRA 21-1), and to define the diagnostic sensitivity and specificity of IL-3 in NSCLC. In this study, the serum level of tumour markers was measured in 34 patients with NSCLC and in 20 healthy subjects. Serum samples were drawn before surgery and 10, 30, 90, 180 and 270 days after surgery. IL-3 and CEA were assayed using ELISA system and CYFRA 21-1 was measured by radioimmunoassay (RIA). Preoperative level of IL-3 was significantly increased in cancer patients relative to the control group. Concentrations of tumour markers were decreased after surgery and then increased (IL-3 and CEA) during chemio- or radiotherapy. The diagnostic sensitivity of IL-3 was 44% and the diagnostic specificity--85%. This investigation is one of the first studies assessing serum levels of IL-3 in the cancer patients. These results suggest that IL-3 may be useful in diagnostics of NSCLC, but this subject needs further studies.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Combined Modality Therapy; Female; Humans; Interleukin-3; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Monitoring, Physiologic; Sensitivity and Specificity

Recently developed tissue polypeptide specific antigen (TPS) and CYFRA 21-1 assays determine the soluble cytokeratin 18 and 19 fragments, respectively, in serum. The authors compared the value of TPS, CYFRA 21-1, and carcinoembryonic antigen (CEA) for the diagnosis, staging, prognosis, and monitoring of patients with nonsmall cell lung carcinoma (NSCLC).. The study included 85 patients with benign lung diseases and 94 patients with NSCLC. TPS, CYFRA 21-1, and CEA serum levels were measured with commercial kits.. The following were demonstrated: 1) CYFRA 21-1 and TPS levels, but not CEA levels, differed significantly between NSCLC patients with operable disease (Stages I-IIIA) and those with inoperable disease (Stages IIIB-IV). 2) The correlation coefficient between CYFRA 21-1 and TPS increased with the progression of NSCLC from Stages I-IIIA (r = 0.41, P = 0.04) to Stages IIIB-IV (r = 0.70, P < 0.001). 3) Multivariate analysis identified TPS and CYFRA 21-1 as significant predictors of survival, with relative risks of 2.57 (P = 0.001) and 2.05 (P = 0.01), respectively. For cases in which both cytokeratin markers were positive, the relative risk was 6.4 (P < 0.0001) compared with cases in which both were negative. 4) For the group with inoperable disease, the combined use of TPS and CYFRA 21-1 allowed for the definition of 3 sets of patients with significantly different median survival times (14.3 months vs. 7.4 months vs. 2.6 months). 5) The percentages of marker evaluations concordant with results of clinical assessments of response to therapy were 75.0%, 72.2%, and 61.1% for CYFRA 21-1, TPS, and CEA, respectively.. These findings suggest that, for NSCLC patients, CYFRA 21-1 and TPS are significant prognostic factors and effective monitors of therapy. The combined use of these cytokeratin markers may provide additional information for prognosis.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Evaluation Studies as Topic; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Monitoring, Physiologic; Neoplasm Staging; Peptides; Predictive Value of Tests; Prognosis; Prospective Studies

Serum tumor markers may be helpful in early diagnosis of cancer, in the initial assessment of the extent of the disease, and in monitoring the tumor growth or tumor volume reduction once cancer has been diagnosed and treatment started. Recent studies have focused on a new family of markers -hematopoietic growth factors, especially on granulocyte-macrophage colony-stimulating factor (GM-CSF). A number of investigations have shown autologous production of GM-CSF in various human cell lines derived from melanoma, gastric or ovarian cancer, and in certain tumors of nonhematopoietic origin. In this study serum level of GM-CSF was measured using a sensitive sandwich ELISA system in 34 patients with non-small cell lung cancer (NSCLC) before and 10, 30, 90, 180 and 270 days after surgical operation. Additionally common accepted tumor markers such as CEA and CYFRA 21.1 were also assayed. Preoperative level of GM-CSF was significantly increased in cancer patients relative to the normal sera (p < 0.02). Concentration of GM-CSF and CYFRA 21.1 were decreased on 10th day, but CEA on 30th day after surgical treatment, although upon comparison of pre- and postoperative tumor markers serum levels significant difference was observed for CYFRA 21.1 (p < 0.05). Levels of GM-CSF were increased in 85%, CEA in 62% and CYFRA 21.1 in 51%. The diagnostic sensitivity and serum levels of GM-CSF were related to the stage of the disease and the combined use of two markers increased the sensitivity compared with the use of only one. These results suggest that GM-CSF, especially in the combination with CYFRA 21.1., may be useful in the diagnostic and monitoring of patients with NSCLC.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Monitoring, Physiologic; Postoperative Care

We evaluated the use of two tumour markers Cyfra 21.1 and tissue polypeptide antigen (TPA) for disease monitoring. Assessment of response to WHO criteria was compared to response assessment according to changes in the tumour marker levels. The criteria defined for marker response were a 65% decrease for a partial response and a 40% increase for progressive disease. When response evaluations with a positive lead time were included, 72% of 115 evaluations for Cyfra 21.1 and 59% of 107 evaluations for TPA yielded the same result. Most discordant evaluations were caused by those evaluations whereby the patient achieved a partial response according to the WHO criteria and had normalisation of the marker. Less cases with a positive lead time, more negative lead times, and more patients with progressive disease without an increase of the marker were seen with TPA compared to Cyfra 21.1. In conclusion, Cyfra 21.1 follows the changes in the tumour load better than TPA. Rising levels of both markers nearly always indicate disease progression, and such knowledge easily obtained may prevent the continuation of ineffective treatment.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Disease Progression; Female; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Peptides; Tissue Polypeptide Antigen

Cytokeratins, the intermediate filaments, are expressed by many epithelial cells. Immunohistochemistry revealed the presence of cytokeratin-19 both in bronchial epithelium and in lung cancer cells. The aim of our study was to establish the value of serum cytokeratin-19 estimation by immunoenzymatic assay (Enzymun Cyfra 21-1, Boehringer Mannheim) in the patients (pts) with lung cancer (Ic). 153 pts (104 men, 49 women, median age 50 years) entered this study. The group consisted of 37 pts with benign lung diseases (control group), 56 pts with squamous cell Ic, 37 pts with small cell Ic and 23 with adenocarcinoma. Cut off value was determined at 4 ng/ml, with 96% of specificity. Elevated cytokeratin-19 values were found in 41% of pts with lung cancer (45% of squamous cell Ic, 39% of adenocarcinoma and 35% of small cell Ic). Median cytokeratin-19 values were 2.2 ng/ml in the control group, 3.4 ng/ml in squamous cell Ic, 3.3 ng/ml adenocarcinoma and 2.9 in small cell Ic. Cytokeratin-19 elevation was observed more often in non small cell Ic pts with advanced disease, stage III--46%, stage IV--50% than in early stages (I + II)--34%. In small cell Ic pts the frequency of cytokeratin-19 elevation was 20% in limited disease versus 45% in extensive disease. We conclude that cytokeratin estimation is not valuable in the recognition of histologic type of lung cancer, although elevated levels are seen more often in squamous cell Ic. Cytokeratin-19 estimation may be also helpful in lung cancer staging.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Biomarkers, Tumor; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Peptide Fragments

The serum levels of cytokeratin-19 were measured in 116 patients--96 with NSCLC and 20 with non-malignant lung diseases. The reference group consisted of 60 healthy volunteers--30 smokers and 30 non-smokers. Significantly elevated Cyfra 21-1 values were observed in NSCLC. There was not a correlation between of cytokeratin-19 serum levels and histologic types of lung carcinoma. Cyfra 21-1 concentrations generally increased with stage of diseases and the highest were in patients with evidence of distant metastases. In NSCLC, the distribution of Cyfra 21-1 varied significantly according to the performance status of NSCLC patients.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Peptide Fragments

We evaluated the newly developed tumor marker assay kit, "Centocor CYFRA 21-1", an immunoradiometric assay (IRMA) kit for determining the serum cytokeratin 19 fragment using the sera of healthy subjects, patients with benign lung diseases and patients with lung cancer. The assay procedure is simple and based on the one-step IRMA system. There were no problems in reproducibility, dilution test and recovery test. The minimum detectable dose was 0.3 ng/ml. The antigen measured by this kit was immunologically cross-reactive with tissue polypeptide antigen (TPA) and CYFRA 21-1 concentration was closely correlated with TPA concentration in the patient's serum (r = 0.86, p < 0.01). The cut-off value of serum CYFRA 21-1 based on the assay results of this kit was calculated to be 1.6 ng/ml from the receiver operating characteristic curve. Three of 47 healthy subjects (6.4%) and 9 of 30 patients with benign lung diseases (30.0%) showed a concentration over the cut-off value. By contrast, serum CYFRA 21-1 concentration was elevated in 31 of 50 patients with lung cancer (62.0%), 11 of 13 squamous cell carcinoma patients (84.6%), 8 of 12 small cell carcinoma patients (66.7%), 4 of 7 large cell carcinoma patients (57.1%) and 8 of 18 adenocarcinoma patients (44.4%). In addition, the positive rate of serum CYFRA 21-1 in patients with lung cancer gradually increased with staging of the disease: 50.0% in stage I, 50.0% in stage II, 61.9% in stage III, and 76.9% in stage IV. Thus, our results suggested that the Centocor CYFRA 21-1 kit is a useful assay system for serum cytokeratin 19 fragment as a tumor marker in patients with lung cancer.

Topics: Adult; Aged; Biomarkers, Tumor; Evaluation Studies as Topic; Female; Humans; Immunoradiometric Assay; Keratins; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Reagent Kits, Diagnostic

The emerging clinical relevance of bone marrow micrometastasis has prompted several investigations, using a variety of immunocytochemical approaches. The present study was designed to evaluate some of the variables affecting the immunocytochemical detection of individual epithelial tumor cells in bone marrow. Using an alkaline phosphatase-antialkaline phosphatase staining technique, we evaluated bone marrow aspirates from 358 patients with primary carcinomas of the breast (n = 150), lung (n = 66), prostate (n = 42), or colorectum (n = 100). Individual tumor cells in cytological preparations were detected with monoclonal antibody (MAb) CK2 to the epithelial cytokeratin component 18 (CK18), which has been validated in extensive clinical studies. In addition, the utility of the broad-spectrum MAb A45-B/B3 was explored in this study. The high specificity of MAbs CK2 and A45-B/B3 was supported by analysis of bone marrow from 75 noncarcinoma control patients and by double-marker analysis with MAbs to mesenchymal marker proteins (CD45 and vimentin). In contrast, MAbs E29 and HMFG1, directed to mucin-like epithelial membrane proteins, cross-reacted with hematopoietic cells in 26.7-42.7% of all samples tested. The majority of the 154 positive samples (43.0%) from cancer patients displayed less than 10 CK18-positive cells per 8 x 10(5) marrow cells analyzed. The detection rate, however, was affected by blood contamination of the aspirate, the number of aspirates analyzed, and the number of marrow cells screened per aspiration site. Comparative immunostaining of bone marrow specimens with MAbs CK2 and A45-B/B3 indicated that downregulation of CK18 in micrometastatic carcinoma cells occurs in about 50% of the 172 samples analyzed, regardless of the primary tumor origin.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Alkaline Phosphatase; Antibodies, Monoclonal; Antibody Specificity; Antigens, Neoplasm; Bone Marrow; Breast Neoplasms; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Membrane Glycoproteins; Mucin-1; Mucins; Neoplasm Metastasis; Prostatic Neoplasms; Sensitivity and Specificity

The are few outstanding serous markers in the treatment of bronchial cancer. ACE lacks sensitivity and specificity and cannot be used as a diagnostic marker. It has been described as a marker of tumoral mass, although not in our study. Prognostic significance was observed, but only in a univariate analysis. Sensitivity of SCC TA4 varied between studies. In our population, the ROC curve for SCC TA4 showed poor discrimination potential. NSE was shown to be a useful marker in the treatment of patients with small cell cancers. Cytokeratins are expressed by all bronchial cancers. Cytokeratin 19 is a sub-unit detected in simple epithelia and their neoplastic counterparts. During tumoral cell lysis, certain fragments of this cytokeratin may be liberated. The immunoradiometric assay described here is able to detect fragments of cytokeratin 19 (called Cyfra 21-1) in serum. Our study established a correlation between Cyfra 21-1 levels and the cancer stage for NCPC, but not for CPC. In addition, Cyfra 21-1 concentration may be used as a tumoral mass marker. Patients with high Cyfra 21 levels must undergo special treatment to find remote tumors.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Diagnosis, Differential; Humans; Immunoradiometric Assay; Keratin-19; Keratins; Lung Neoplasms; ROC Curve; Sensitivity and Specificity; Survival Rate

Small cell lung carcinoma (SCLC), a malignancy of neuroendocrine origin, can show varied morphologies and patterns but is typically positive for at least one neuroendocrine marker and almost always for cytokeratins. It is essential to distinguish this tumour due to its characteristic genetic features, aggressive behaviour, propensity for metastasis and responsiveness to chemotherapy. We hereby present a rare case of a pulmonary mass that showed morphological features of an SCLC but lacked cytokeratin expression on biopsy as well as resection specimens. Various cytokeratins were tested on multiple blocks and at different laboratories. A broad differential diagnosis was considered and ruled out including small round blue cell tumours, non-SCLC and metastasis. After performing an extensive work-up to identify the origin of this tumour, it was finally diagnosed as SCLC with expression of neuroendocrine markers synaptophysin and CD56, and intracytoplasmic electron dense neurosecretory granules (250-350 nm) however lacked cytokeratin expression.

Topics: Biomarkers, Tumor; Biopsy; Carcinoma, Neuroendocrine; Carcinoma, Non-Small-Cell Lung; Humans; Keratins; Lung Neoplasms; Small Cell Lung Carcinoma

The aim of the study was to assess the diagnostic value of tumor markers in discriminating between lung cancer and benign chest diseases (BCDs).. There were 322 patients enrolled in this investigation including 180 cases of lung cancer and 142 cases of BCD. Serum neuron-specific enolase (NSE), cancer antigen 125, cancer antigen 19-9, squamous cell carcinoma-related antigen, and cytokeratin fragment 19 (CYFRA 21-1) were compared between different populations, cancer stages, and before and after treatment. Logistic regression and receiver operating characteristic curves were used to evaluate the diagnostic markers.. Both NSE and CYFRA 21-1 were significantly associated with lung cancer. The CYFRA 21-1 showed the best performance, as well as its combinations, for lung cancer diagnosis. It also showed significant change 6 months after radical surgery in lung cancer patients.. The marker CYFRA 21-1 could be developed as an adjuvant marker for the early diagnosis of lung cancer and as a prognostic marker for lung cancer treatment.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; CA-125 Antigen; CA-19-9 Antigen; Humans; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase; Sensitivity and Specificity

The presence of tumor abnormal protein (TAP) is closely related to the development and progression of many cancers. During metabolism, cancer cells can release complicated abnormal glycoproteins as well as calcium histone proteins which constitute TAP. In essence, TAP results from the glycosylation changes related to cancer cells. In this study, TAP and tumor markers were assessed for their diagnostic value in lung cancer. The serum levels of TAP, pro-gastrin-releasing peptide, carcinoembryonic antigen, CYFRA 21-1 and CA72-4 in the patients with lung cancer were significantly higher than in those with benign lung diseases. TAP combined with CYFRA 21-1 and CA72-4 or with CYFRA 21-1, CA72-4 and squamous cell carcinoma antigen could further improve the sensitivity of lung cancer diagnosis and is suitable for lung cancer screening in both normal and high-risk populations.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Humans; Keratin-19; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase; Sensitivity and Specificity

Immunotherapy is currently part of the standard of care for patients with advanced-stage non-small cell lung cancer (NSCLC). However, many patients do not respond to this treatment, therefore combination strategies are being explored to increase clinical benefit. The PEMBRO-RT trial combined the therapeutic programmed cell death 1 (PD-1) antibody pembrolizumab with stereotactic body radiation therapy (SBRT) to increase the overall response rate and study the effects on the tumor microenvironment (TME).. Here, immune infiltrates in the TME of patients included in the PEMBRO-RT trial were investigated. Tumor biopsies of patients treated with pembrolizumab alone or combined with SBRT (a biopsy of the non-irradiated site) at baseline and during treatment were stained with multiplex immunofluorescence for CD3, CD8, CD20, CD103 and FoxP3 for lymphocytes, pan-cytokeratin for tumors, and HLA-ABC expression was determined.. The total number of lymphocytes increased significantly after 6 weeks of treatment in the anti-PD-1 group (fold change: 1.87, 95% CI: 1.06 to 3.29) and the anti-PD-1+SBRT group (fold change: 2.29, 95% CI: 1.46 to 3.60). The combination of SBRT and anti-PD-1 induced a 4.87-fold increase (95% CI: 2.45 to 9.68) in CD103. This explorative study shows that that lymphocyte infiltration in general, instead of the infiltration of a specific lymphocyte subset, is associated with response to therapy in patients with NSCLC.Furthermore, anti-PD-1+SBRT combination therapy induces an immunological abscopal effect in the TME represented by a superior infiltration of cytotoxic T cells as compared with anti-PD-1 monotherapy.

Topics: Antibodies, Monoclonal, Humanized; Carcinoma, Non-Small-Cell Lung; Forkhead Transcription Factors; Humans; Keratins; Lung Neoplasms; Tumor Microenvironment

A 67-year-old woman was referred to our hospital for cough and fever. Chest computed tomography (CT) showed some masses showing slightly enhanced effect in the pericardium. FDG-PET showed the accumulation of FDG in the masses. Thoracoscopic surgical biopsy was performed to establish the diagnosis. The histological study showed proliferation of short spindle-shaped cells surrounded by lymphocyte, and the spindle cells were immunohistochemically positive for cytokeratin AE1/AE3, WT-1, D2-40, CAM5.2, intelectin-1 and negative for CEA, TTF-1, napsin A, claudin-4, calretinin, MUC4, PAX8, CD30. These findings were compatible with epithelial pericardial malignant mesothelioma.

Topics: Aged; Calbindin 2; Claudin-4; Female; Fluorodeoxyglucose F18; Heart Neoplasms; Humans; Keratins; Lung Neoplasms; Mediastinal Neoplasms; Mesothelioma; Mesothelioma, Malignant; Thymus Neoplasms

Mice have served as an excellent model to understand the etiology of lung cancer for years. However, data regarding dual-stage carcinogenesis of lung squamous cell carcinoma (SCC) remain elusive. Therefore, we aim to develop pre-malignant (PM) and malignant (M) lung SCC in vivo using N-nitroso-tris-chloroethylurea (NTCU). BALB/C mice were allotted into two main groups; PM and M groups which received treatment for 15 and 30 weeks, respectively. Then, the mice in each main group were allotted into three groups; control, vehicle, and cancer (n = 6), which received normal saline, 70% acetone, and 0.04 M NTCU by skin painting, respectively. Histopathologically, we discovered a mix of hyperplasia, metaplasia, and dysplasia lesions in the PM group and intracellular bridge; an SCC feature in the M group. The M group was positive for cytokeratin 5/6 protein which confirmed the lung SCC subtype. We also found significantly higher (P < 0.05) epithelium thickness in the cancer groups as compared to the vehicle and control groups at both the PM and M. Overall, this study discovered that NTCU is capable of developing PM and M lung SCC in mice model at appropriate weeks and the vehicle group was suggested to be adequate as control group for future research.

Topics: Animals; Biomarkers, Tumor; Carcinogens; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Carmustine; Disease Models, Animal; Epithelium; Female; Immunohistochemistry; Keratins; Lung Neoplasms; Mice; Mice, Inbred BALB C

Topics: Aged, 80 and over; Antineoplastic Combined Chemotherapy Protocols; Biopsy; Bone Marrow; Esophageal Neoplasms; Gene Expression; Humans; Immunohistochemistry; Keratins; Leukemia; Lung Neoplasms; Male; Neoplasms, Second Primary; Radiotherapy

Liquid biopsy analyzes the current status of primary tumors and their metastatic regions. We aimed to develop an optimized protocol for single-cell sequencing of floating tumor cells (FTCs) in pleural effusion as a laboratory test. FTCs were enriched using a negative selection of white blood cells by a magnetic-activated cell sorting system, and CD45-negative and cytokeratin-positive selection using a microfluidic cell separation system with a dielectrophoretic array. The enriched tumor cells were subjected to whole-genome amplification (WGA) followed by genome sequencing. The FTC analysis detected an EGFR exon 19 deletion in Case 1 (12/19 cells, 63.2%), and EML4-ALK fusion (17/20 cells, 85%) with an alectinib-resistant mutation of ALK (p.G1202R) in Case 2. To eliminate WGA-associated errors and increase the uniformity of the WGA product, the protocol was revised to sequence multiple single FTCs individually. An analytical pipeline, accurate single-cell mutation detector (ASMD), was developed to identify somatic mutations of FTCs. The large numbers of WGA-associated errors were cleaned up, and the somatic mutations detected in FTCs by ASMD were concordant with those found in tissue specimens. This protocol is applicable to circulating tumor cells analysis of peripheral blood and expands the possibility of utilizing molecular profiling of cancers.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Anaplastic Lymphoma Kinase; Antineoplastic Agents; Carbazoles; Cell Separation; Crizotinib; DNA; Drug Resistance, Neoplasm; Exons; Female; Gene Amplification; Gene Deletion; Gene Expression Profiling; Genes, erbB-1; High-Throughput Nucleotide Sequencing; Humans; Immunomagnetic Separation; Keratins; Leukocyte Common Antigens; Liquid Biopsy; Lung Neoplasms; Male; Middle Aged; Mutation; Neoplastic Cells, Circulating; Oncogene Proteins, Fusion; Piperidines; Pleural Effusion, Malignant; Protein Kinase Inhibitors

The TSA Opal multiplex immunohistochemistry (mIHC) protocol (PerkinElmer) has been used to characterize immune infiltration in human cancers. This technique allows multiple biomarkers to be simultaneously stained in a single tissue section, which helps to elucidate the spatial relationship among individual cell types. We developed and optimized two improved mIHC protocols for a 7-color panel containing 6 biomarkers (CD3, CD8, CD163, PD-L1, FoxP3, and cytokeratin (CK)) and DAPI. The only difference between these two protocols was the staining sequence of those 6 biomarkers as the first sequence is PD-L1/CD163/CD8/CK/CD3/FoxP3/DAPI and the second sequence is FoxP3/CD163/CD8/CK/CD3/PD-L1/DAPI. By comparing PD-L1/FoxP3 staining in mIHC and singleplex PD-L1/FoxP3 staining on the adjacent slide, we demonstrated that the staining sequence does not affect the staining intensity of individual biomarkers as long as a proper antigen retrieval method was used. Our study suggests that use of an antigen retrieval buffer with higher pH value (such as Tris-EDTA pH9.0) than that of the stripping buffers (such as citrate buffer pH6.0) is helpful when using this advanced mIHC method to develop panels with multiple biomarkers. Otherwise, individual biomarkers may exhibit different intensities when the staining sequence is changed. By using this protocol, we characterized immune infiltration and PD-L1 expression in head and neck squamous cell carcinoma (HNSCC), breast cancer (BCa), and non-small cell lung cancer (NSCLC) specimens. We observed a statistically significant increase in CD3+ cell populations within the stroma of NSCLC as compared to BCa and increased PD-L1+ tumor cells in HNSCC as opposed to BCa.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; B7-H1 Antigen; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Non-Small-Cell Lung; CD3 Complex; CD8 Antigens; Female; Forkhead Transcription Factors; Head and Neck Neoplasms; Humans; Immunohistochemistry; Indoles; Keratins; Lung Neoplasms; Lymphocytes, Tumor-Infiltrating; Receptors, Cell Surface; Squamous Cell Carcinoma of Head and Neck

We previously proposed an immune cell score (tumour node metastasis (TNM)-Immune cell score) classifier as an add-on to the existing TNM staging system for non-small cell lung cancer (NSCLC). Herein, we examined how to reliably assess a tertiary lymphoid structure (TLS) score to refine the TNM staging system.. Using immunohistochemistry (CD8/cytokeratin), we quantified TLS in resected NSCLC whole-tumour tissue sections with three different scoring models on two independent collections (total of 553 patients). In a pilot setting, NanoString gene expression signatures were analysed for associations with TLS.. The number of TLSs significantly decreased in stage III patients as compared to stage II. The TLS score was an independent positive prognostic factor, regardless of the type of (semi)-quantification strategy used (four-scale semi-quantitative; absolute count of total TLS; subpopulation of mature TLS) or the endpoint (disease-specific survival; overall survival; time to recurrence). Subgroup analyses revealed a significant prognostic impact of TLS score within each pathological stage, patient cohort and main histological subtype. Targeted gene expression analysis showed that high TLS levels were associated with the expression of B cell and adaptive immunity genes/metagenes including tumour inflammation signature.. The TLS score increases the prognostic power in each pathological stage and hence has the potential to refine TNM staging in resected NSCLC.

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; CD8 Antigens; Cohort Studies; Follow-Up Studies; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphocytes, Tumor-Infiltrating; Neoplasm Staging; Norway; Prognosis; Research Design; Tertiary Lymphoid Structures; Transcriptome; Tumor Microenvironment

为更好地显示肺腺癌中弹力纤维的变化及其与肿瘤细胞的关系，应用全自动免疫组织化学和全自动特殊染色平台对30例肺腺癌组织细胞进行角蛋白（CK）7和弹力纤维双重染色，结果显示在双染切片中CK7免疫组织化学的特异度和灵敏度与单独染色基本没有差异，两者相结合除可以特异性显示胸膜受侵的情况，还观察到多数厚壁血管弹力纤维的增生和血管壁的重构，以及更多CK7和弹力纤维单独染色难以观察到的肿瘤细胞早期浸润血管壁的现象。.

Topics: Adenocarcinoma of Lung; Automation; Biomarkers, Tumor; Diagnosis, Differential; Elastic Tissue; Humans; Keratin-7; Keratins; Lung Neoplasms; Staining and Labeling

A case of a primary lung carcinoma with histologic and immunohistochemical features of a mammary carcinoma is presented. The patient is a 72-year-old man who presented with symptoms of cough and dyspnea. Diagnostic imaging showed a bronchial tumor in the left lower lobe that was surgically resected by a left lower lobectomy. The tumor was characterized by a homogenous cellular proliferation composed of small to medium-sized cells with round nuclei and inconspicuous nucleoli. Multiple immunohistochemical stains were performed, and the tumor was notably positive for estrogen receptor, progesterone receptor, GATA3, and pan-keratin, while molecular analysis showed somatic mutation in

Topics: Aged; Biomarkers, Tumor; Carcinoma; DNA-Binding Proteins; GATA3 Transcription Factor; Humans; Keratins; Lung Neoplasms; Male; Mutation; Receptors, Estrogen; Receptors, Progesterone; Transcription Factors

Fibroblast growth factor receptor 1 (FGFR1) is frequently amplified in human small-cell lung cancer (SCLC), but its contribution to SCLC and other lung tumors has remained elusive. Here, we assess the tumorigenic capacity of constitutive-active FGFR1 (FGFR1

Topics: Adenocarcinoma of Lung; Animals; Bronchi; Cell Transformation, Neoplastic; Cisplatin; Drug Resistance, Neoplasm; Gene Expression Regulation, Neoplastic; Humans; Integrases; Keratins; Lung Neoplasms; Mice; Mutation; Nasal Cavity; Neurosecretory Systems; Oncogenes; Pulmonary Alveoli; Receptor, Fibroblast Growth Factor, Type 1; Retinoblastoma Protein; Small Cell Lung Carcinoma; Tumor Suppressor Protein p53

The role of Cytokeratin fraction 21-1 (CYFRA 21-1) as a tumour marker for head and neck cancer is still a matter of research. The aim of the present study was to evaluate the clinical impact of CYFRA 21-1 for patients with oropharyngeal squamous cell carcinoma (OSCC).. Data of 180 patients with an initial diagnosis of OSCC of any stage between 2003 and 2017 were retrospectively analysed regarding the association between pretherapeutic CYFRA 21-1 levels, clinical characteristics, overall and disease-free survival. Additionally, the potential of CYFRA 21-1 for the detection of recurrent disease in the follow-up was evaluated. The cut-off value was set at 3.3 ng/ml. The median follow-up time was 2.85 years.. A significant correlation of the CYFRA 21-1 concentration at the time of diagnosis and the N-stage was detected (p = 0.01). Patients with CYFRA 21-1 levels > 3.3 ng/ml at first diagnosis showed a significantly shorter overall survival. In the case of disease-progression, a significant increase of CYFRA 21-1 value was found compared to post-therapeutic CYFRA 21-1 levels (9.1 ng/ml versus 5.1 ng/ml; p < 0.01). CYFRA 21-1 level after treatment showed only a low sensitivity of 32% and a specificity of 78% for tumour recurrence.. CYFRA 21-1 correlates with the tumour stage and, therefore, the survival of OSCC patients. Posttreatment CYFRA21-1 seems not to be a suitable predictor of tumour recurrence in the further course of the disease. However, a sudden increase of CYFRA 21-1 during follow-up may indicate a tumour recurrence in the individual patient.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Lung Neoplasms; Neoplasm Recurrence, Local; Retrospective Studies; Sensitivity and Specificity; Squamous Cell Carcinoma of Head and Neck

There is currently no definite clinical implication for the subtypes of lung squamous cell carcinoma according to the 2015 WHO classification. This study aimed to investigate postoperative recurrence of the two major subtypes of lung squamous cell carcinoma: keratinizing squamous cell carcinoma (KSCC) and nonkeratinizing squamous cell carcinoma (NKSCC).. We identified the patients with KSCC and NKSCC who had undergone complete resection in Shanghai Chest Hospital between April 2015 and June 2016. Disease-free survival (DFS) was compared using Kaplan-Meier statistical analysis. Variables selected by univariate analysis were evaluated in multivariate analysis using the Cox proportional hazard model.. A total of 334 patients included 231 (69.2%) cases with KSCC and 103 (30.8%) cases with NKSCC. There were more smokers in keratinizing than nonkeratinizing subtype (84.8% versus 72.8%, p = 0.009). The percentage of stage Ⅲ was higher in NKSCC than that in KSCC (35% versus 22.9%, p = 0.012). The 2-year DFS rates of stage Ⅰ, stage Ⅱ and stage Ⅲ were 90.1%, 66.4% and 37.7% in KSCC, 83.3%, 67.7% and 52.8% in NKSCC, respectively. There were no significant differences of 2-year DFS rates between KSCC and NKSCC. Furthermore, KSCC and NKSCC had no significant differences in recurrence patterns and metastatic sites.. There were no significant differences of postoperative recurrence between KSCC and NKSCC.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Chemotherapy, Adjuvant; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasm Staging; Radiotherapy, Adjuvant

Three cases of primary carcinomas of the lung each with an extensive osteoclast-like giant cell component are presented. The patients are 3 men between the ages of 58 and 67 years (average, 62.5 years) who presented with nonspecific symptoms. A history of malignancy, infectious, or granulomatous disease was negative in all the patients. Diagnostic imaging disclosed the presence of a large intrapulmonary mass; in 1 case in the right upper lobe and in 2 cases in the right lower lobe. Surgical resection via lobectomy was performed in the 3 patients. Grossly, the tumors were described as soft, friable intrapulmonary masses, reddish in color, and measuring from 6 to 13 cm in largest diameter. Histologically, the tumors were each characterized by the extensive presence of a multinucleated osteoclast-like giant cell component, which represented approximately 80% of the tumor mass. The osteoclast-like giant cell component was admixed with a sarcomatoid carcinoma in 2 cases and an adenocarcinoma in 1 case. Immunohistochemistry showed that the osteoclast-like giant cells were positive for CD-68, cathepsin K, and histone H3, whereas the carcinoma component was positive for keratin, thyroid transcription factor-1, and histone H3 (patchy). Molecular studies were performed in 2 patients with negative results. Clinical follow-up was obtained in 2 patients; 1 died 14 months after initial diagnosis, whereas 1 remains alive 6 months after initial diagnosis. One patient was lost to follow-up. The current neoplasms represent an unusual type of lung carcinoma that needs highlighting as a separate type from conventional giant cell carcinoma.

Topics: Aged; Antigens, CD; Biomarkers, Tumor; Carcinoma, Giant Cell; Giant Cells; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Osteoclasts; Retrospective Studies; Thyroid Nuclear Factor 1

Most cancer diagnoses rely on biomarkers detection. This could be improved if directly conducted in suspicious cancer spots, preventing the need for biopsy. Lung cancer remains a perfect study-case for such a development, as it is generally detected at advanced stage and is in the need for early diagnosis techniques. To this aim, we have designed a minimally invasive catheter-embedded biosensor. It combines a specific grating structure photo-imprinted in a telecommunication-grade optical fiber and an overlay made of a thin metal coating on which receptors are grafted, yielding plasmonic coupling. Our optrode targets a type of cytokeratins, overexpressed at the surface of cancer cells. It was assayed ex vivo in resected lung tissues collected from a dozen of patients. Biosensing responses were confirmed by immunohistochemistry, conducted on the same samples. In addition to accurate biosensing, our gratings inherently enable force-sensing features, which also allow a fine positioning of the probe in the tissue. Finally, the in vivo navigation of the bronchoscope-embedded sensor was validated into pig lungs. These achievements are a critical milestone towards the development of this micro/nano biosensor as a cost-effective and weakly invasive diagnostic tool for applications in areas of critical access such as brain, liver or prostate.

Topics: Animals; Biosensing Techniques; Cell Line, Tumor; Fiber Optic Technology; Gene Expression Regulation, Neoplastic; Humans; Keratins; Lung; Lung Neoplasms; Optical Fibers; Surface Plasmon Resonance; Swine

Circulating tumor cells (CTCs) are putative markers of tumor prognosis and may serve to evaluate patient's response to chemotherapy. CTCs are often detected as single cells but infrequently as clusters and are indicative of worse prognosis. In this study, we developed a short-term culture of nucleated blood cells which was applied to blood samples from breast, lung, esophageal and bladder cancer patients. Clusters of different degrees of compactness, classified as very tight, tight and loose were observed across various cancer types. These clusters show variable expression of cytokeratins. Cluster formation from blood samples obtained during the course of chemotherapy was found to be associated with disease progression and shorter overall survival. The short-term cultures offer a robust and highly reliable method for early prediction of treatment response in different cancer types.

Topics: Antineoplastic Agents; Breast Neoplasms; Disease Progression; Esophageal Neoplasms; Female; Humans; Kaplan-Meier Estimate; Keratins; Lung Neoplasms; Neoplastic Cells, Circulating; Prognosis; Tumor Cells, Cultured; Urinary Bladder Neoplasms

A 10-year-old female American Pit Bull dog was diagnosed with metastatic undifferentiated carcinoma of the scapula. Immunohistochemistry showed positive immunoexpression for cytokeratins (AE1/AE3, 34BE12, CK7) and vimentin, confirming squamous cell carcinoma. No evidence of nodules was found in the complete physical examination and imaging procedures conducted. The patient was diagnosed with carcinoma of unknown primary origin. Amputation and adjuvant chemotherapy with doxorubicin and piroxicam were performed, but the patient died of respiratory failure after 737 days of diagnosis. Necropsy confirmed undifferentiated carcinoma infiltrating the lungs and kidneys, and showing the same immunoexpression as the tumor in the scapula. Amputation associated with chemotherapy extended the overall survival time of this patient.

Topics: Amputation, Surgical; Animals; Biomarkers, Tumor; Bone Neoplasms; Carcinoma, Squamous Cell; Dogs; Drug Therapy; Female; Immunohistochemistry; Keratins; Kidney Neoplasms; Lung Neoplasms; Neoplasms, Unknown Primary; Scapula; Vimentin

An increasing body of evidence has demonstrated that microRNA (miR) deregulation serves pivotal roles in tumor progression and metastasis. However, the function of miR‑379 in lung cancer remains understudied, particularly in non‑small cell lung cancer (NSCLC). Bioinformatics and luciferase reporter analyses confirmed that conserved helix‑loop‑helix ubiquitous kinase (CHUK) is a target of miR‑379, which may directly bind to the 3'‑untranslated region of CHUK and significantly downregulate its expression in NSCLC cells. Transwell assays were used to evaluate the role of miR‑379 in cell migration and invasion, and western blotting was used to address the association between miR‑379 and epithelial‑mesenchymal markers, including E‑cadherin, cytokeratin and Vimentin. In the present study, miR‑379 expression in NSCLC tissues and cell lines was downregulated, which may be associated with the poor survival of patients with NSCLC. miR‑379 may act as a tumor suppressor in NSCLC, potentially by suppressing cell growth and proliferation, delaying G1‑S transition, enhancing cell apoptosis and suppressing NSCLC cell migration and invasion. Furthermore, it was also observed that CHUK may function as an oncogene, and downregulation of CHUK induced by miR‑379 may partially rescue the malignant characteristics of tumors, indicating that miR‑379 may be suppressed in tumorigenesis. The overexpression of miR‑379 may prevent the growth of NSCLC tumors via CHUK suppression and the downstream nuclear factor‑κB pathway. The results of the present study demonstrated that miR‑379 may act as a tumor suppressor, and may constitute a potential biomarker and a promising therapeutic agent for the treatment for NSCLC.

Topics: Antigens, CD; Base Sequence; Cadherins; Carcinoma, Non-Small-Cell Lung; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Humans; I-kappa B Kinase; Keratins; Lung Neoplasms; MicroRNAs; NF-kappa B; Oligoribonucleotides; Signal Transduction; Tissue Culture Techniques; Vimentin

The goal of this work was to develop an LC-MRM assay for the quantitative analysis of a set of established and diagnostically important cytokeratin (CK) markers used in cancer diagnosis, prognosis, and therapy monitoring. Second, the potential of this assay in lung cancer diagnosis through pleural effusion (PE) analysis was examined.. A multiplexed MRM assay was developed for 17 CKs and their select caspase-cleaved fragments. Isotope-labeled standard peptides were used for high assay specificity and absolute peptide quantitation; with robust standard-flow LC coupled to a latest-generation triple-quadrupole instrument for high sensitivity. The potential clinical applicability was demonstrated by the analysis of 118 PE samples.. The MRM assay was evaluated for endogenous detection, linearity, precision, upper and lower limits of quantification, selectivity, reproducibility and peptide stability, and is generally applicable to any epithelial cancer study. A set of 118 patients with known pathologies allowed us to define the range of CK levels in clinical PE samples. Specific CKs were able to differentiate cancer-related PEs from those caused by benign ailments. In addition, they allowed to differentiate between PEs from subjects with small cell lung cancer versus non-small cell lung carcinoma, and to further differentiate the latter into its two subtypes, adenocarcinoma and squamous cell carcinoma.. An MRM-based CK assay for carcinoma studies can differentiate between the three lung cancer histological types using less-invasive PE sampling providing potential therapy-guiding information on patients that are inoperable.

Topics: Amino Acid Sequence; Biomarkers, Tumor; Humans; Keratins; Lung Neoplasms; Mass Spectrometry; Pleural Effusion, Malignant; Prognosis; Proteomics

Topics: Aged, 80 and over; Biomarkers, Tumor; Humans; Keratins; Lung Neoplasms; Male; Mesothelioma; Mesothelioma, Malignant; Pleural Neoplasms

This study aimed to investigate the significance of lymph node micrometastasis (LNMM) in the lung cancer nodal categories.. Between 1 January 2009 and 31 December 2013, 589 patients with suspected c-stage 1 and p-T1-2aN0-1M0 lung adenocarcinoma were enrolled in this study. The study evaluated LNMM with cytokeratin (AE1/AE3) and transcription factor-1 (TTF1) (8G7G3/1) expression by immunohistochemistry. Recurrence-free survival (RFS) and overall survival (OS) were compared among the T1-2aN0-1M0 patients stratified by the new N categories.. From 589 patients, 7892 removed lymph nodes were examined, and LNMM was observed in 55 (9.3%) of the patients. The patients without LNMM or N1 had the best RFS (5-year rate: 80% vs 25%; P < 0.001) and OS (5-year rate: 87% vs 43%; P < 0.001), followed by the patients with LNMM, compared with those in the N1 category (RFS: 5-year rate, 25% vs 8%; P = 0.010; OS: 5-year rate, 43% vs 20%; P = 0.009). Similarly, this trend was observed when patients were subdivided into the T1 and T2a categories. Multivariate analysis showed that the new N categories with the addition of LNMM were an independent prognostic factor. This result also was noticed in all subgroups.. The findings showed LNMM to be clinically significant as a risk factor for lung cancer. Clinicians should consider LNMM when estimating N categories to determine prognosis and the best treatment strategy.

Topics: Adenocarcinoma; Aged; Disease-Free Survival; Female; Humans; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Micrometastasis; Neoplasm Staging; Survival Rate; Thyroid Nuclear Factor 1

Solitary pulmonary capillary hemangioma (SPCH) is a rare lung tumor typically presenting as pure or part-solid ground-glass nodules (GGNs) on computed tomography (CT), which clinically resembles early lung cancer.. In addition to 10 recently diagnosed patients with SPCH, 71 benign lung nodules that were surgically resected between January 2013 and December 2017 were reviewed by thoracic pathologists to identify any previously unrecognized SPCH cases. Finally, 6 tumors (8.5%; 6/71) were determined to be SPCH. Elastic fiber stain (orcein stain) as well as immunohistochemistry for cytokeratin, CD31, and thyroid transcription factor 1 were performed for confirmation. Clinical and radiological data were analyzed.. All 16 SPCH lesions were unrecognized or misdiagnosed by general pathologists. The SPCH ranged in size from 3 to 15 mm, and a predominance among women (68.8%; 11/16) was noted. Pathologically, all SPCH lesions were nodular with a higher vascular density than the adjacent lung tissue. Decreased cytokeratin staining and disrupted elastic fibers were clearly observed in all SPCH lesions.. SPCH lesions mimic early lung cancer on CT; they are largely unrecognized by general pathologists and are diagnosed as other nonspecific benign lesions. With careful histologic examination, SPCH can be successfully diagnosed using cytokeratin/CD31 immunohistochemistry and elastic fiber staining.

Topics: Adult; Aged; Diagnosis, Differential; Diagnostic Errors; Elastic Tissue; Female; Hemangioma, Capillary; Humans; Keratins; Lung; Lung Neoplasms; Male; Middle Aged; Sex Factors; Solitary Pulmonary Nodule; Tomography, X-Ray Computed; Tumor Burden

Topics: Elastic Tissue; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Neoplasm Invasiveness; Pleura; Staining and Labeling

Circulating tumor cell (CTC) as an important component in "liquid biopsy" holds crucial clinical relevance in cancer prognosis, treatment efficiency evaluation, prediction and potentially early detection. Here, we present a Fiber-optic Array Scanning Technology (FAST) that enables antigen-agnostic, size-agnostic detection of CTC. By immunofluorescence staining detection of a combination of a panel of markers, FAST technology can be applied to detect rare CTC in non-small cell lung cancer (NSCLC) setting with high sensitivity and specificity. In combination with Automated Digital Microscopy (ADM) platform, companion markers on CTC such as Vimentin and Programmed death-ligand 1 (PD-L1) can also be analyzed to further characterize these CTCs. FAST data output is also compatible with downstream single cell picking platforms. Single cell can be isolated post ADM confirmation and used for "actionable" genetic mutations analysis.

Topics: Antibodies, Monoclonal; B7-H1 Antigen; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Early Detection of Cancer; Fiber Optic Technology; Fluorescent Antibody Technique; Fluorescent Dyes; Humans; Keratins; Leukocyte Common Antigens; Leukocytes, Mononuclear; Lung Neoplasms; Mucin-1; Neoplastic Cells, Circulating; Receptor, ErbB-2; Sensitivity and Specificity; Single-Cell Analysis; Vimentin

Primary pericardial malignant mesothelioma (PPM) is a very rare malignancy, with an incidence of less than 0.002% and represents less than 5% of all mesotheliomas. The cause of pericardial mesothelioma is uncertain that differ from pleural mesothelioma which is associated with asbestos exposure. This malignancy is terribly aggressive and has very poor prognosis with less than six months of overall survival. We present a case of a 71-year-old woman who was diagnosed with cardiac tamponade caused by PPM and received chemotherapy with pemetrexed and cisplatin for six months. During two years she was alive without disease progression. To better understand the clinical, pathologic features and treatment outcome of this entity, we reviewed 23 cases described in the English literature from 2009, together with our case, provided a total of 24 cases. Based on this review, we suggest that PPM must be considered in patients who have unexplained massive pericardial effusion and recommend chemotherapy with pemetrexed and cisplatin for the better outcome of PPM.

Topics: Aged; Antineoplastic Agents; Calbindin 2; Cardiac Tamponade; Cisplatin; Drug Therapy, Combination; Female; Humans; Keratins; Lung Neoplasms; Mesothelioma; Mesothelioma, Malignant; Pemetrexed; Pleural Neoplasms; Thorax; Tomography, X-Ray Computed; Treatment Outcome; Ultrasonography; Vimentin

Sphingosylphosphorylcholine induces keratin phosphorylation and reorganization, and increases viscoelasticity of metastatic cancer cells such as PANC-1 cells. However, the mechanism involved in sphingosylphosphorylcholine-induced keratin phosphorylation and reorganization is largely unknown. Sphingosylphosphorylcholine dose- and time-dependently induces the expression of RhebL1. The involvement of RhebL1 in sphingosylphosphorylcholine-induced events including keratin 8 (K8) phosphorylation, reorganization, migration and invasion was examined. Gene silencing of RhebL1 suppressed the sphingosylphosphorylcholine-induced events and overexpression of RhebL1 enhanced those events even without sphingosylphosphorylcholine treatment. We examined whether the G protein function of RhebL1 induces K8 phosphorylation using constitutively active RhebL1Q64L and dominant negative RhebL1D60K. G protein activity of RhebL1 is involved in sphingosylphosphorylcholine-induced K8 phosphorylation. We found that RhebL1 binds and activates AKT1. G protein activity of RhebL1 is involved in the binding and activation of AKT1. MK2206 (AKT inhibitor) and gene silencing of AKT1 inhibited the sphingosylphosphorylcholine-induced events, whereas overexpression of activated-AKT1 induced K8 phosphorylation, reorganization, migration and invasion even without sphingosylphosphorylcholine treatment.The collective results indicate that RhebL1 is involved in sphingosylphosphorylcholine-induced events in A549 lung cancer cells via binding to AKT1 leading to activation of it. These results suggest that suppression of RhebL1 or inhibition of RhebL1's binding to AKT1 might be a novel way that prevents changes in the physical properties of metastatic cancer cells.

Topics: Apoptosis; Biomarkers, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; GTP-Binding Proteins; Humans; Keratins; Lung Neoplasms; Neoplasm Staging; Phosphorylation; Phosphorylcholine; Prognosis; Proto-Oncogene Proteins c-akt; ras Proteins; Sphingosine; Survival Rate; Tumor Cells, Cultured

Pulmonary sarcomatoid carcinoma (PSC) is a rare malignancy.. A total of 69 patients with PSC treated at a single institution in southern China with long-term follow-up were evaluated in this study. We analyzed the clinical characteristics, immunohistochemical profiles, epidermal growth factor receptor mutation status, K-RAS mutation status, treatments, and prognosis.. PSC mainly occurred in young male patients with a history of smoking. Most patients received multimodality treatments and the majority had early-stage disease. The median survival time was 19.1 months, and the 5-year survival rate was 17.4%. The patients without distant metastasis, with normal or higher body mass index (≥18.5), with normal hemoglobin, with smaller tumor size (≤4 cm), and those who received complete resection had significantly better overall survival (P<0.05). The patients with pleomorphic carcinoma had much worse prognosis. In a Cox regression model, M stage, pathology, and having received a complete resection were independent prognostic factors (P<0.05).. PSC is a unique lung malignancy with poor prognosis. Patients receiving complete resection had better prognosis, likely a reflection of early-stage disease. Neither neoadjuvant nor adjuvant chemotherapy improved patient survival for those with early-stage disease. The retrospective design and small sample size limited the generalizability. Future multicenter collaborations may be necessary to determine the optimal treatment.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Carcinoma; Carcinoma, Giant Cell; Carcinosarcoma; Combined Modality Therapy; Disease-Free Survival; ErbB Receptors; Female; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Mucin-1; Nuclear Proteins; Prognosis; Proto-Oncogene Proteins p21(ras); Pulmonary Blastoma; S100 Proteins; Survival Rate; Thyroid Nuclear Factor 1; Transcription Factors; Vimentin; Young Adult

Carcinomas and their metastases often retain the keratin patterns of their epithelial origin, and are therefore useful as lineage-specific markers in diagnostic pathology. Recently, it has become clear that intermediate filaments composed by keratins play a role in modulation of cell proliferation, migration, and possibly cancer invasion, factors impacting prognosis in early stage non-small cell lung cancer (NSCLC).. Tumor tissue from a retrospective Danish cohort of 177 patients with completely resected NSCLC, stage I-IIIA tumors, were analyzed for keratin 7 (K7) and keratin 34βE12 expression by immunohistochemistry and validated in a comparable independent Norwegian cohort of 276 stage I-IIIA NSCLC patients.. Based on keratin 34βE12/K7 expression, three subgroups with significantly different median cancer-specific survival rates were identified (34βE12+/K7+, 168 months vs. 34βE12+/K7+, 73 months vs. 34βE12-/K7+, 30 months; p = 0.0004). In multivariate analysis, stage II-IIIA (HR 2.9), 34βE12+/K7+ (HR 1.90) and 34βE12-/K7+ (HR 3.7), were prognostic factors of poor cancer-specific survival (CSS) (p < 0.001). Validation in the Norwegian cohort confirmed that stage II-IIIA (HR 2.3), 34βE12+/K7+ (HR 1.6), and 34βE12-/K7+ (HR 2.0) were prognostic factors of poor CSS (p < 0.05). Multivariate Cox proportional-hazard analysis demonstrated that 34βE12+/K7 + and 34βE12+/K7 + status was significantly associated with poor overall survival (p < 0.05).. Keratin 34βE12/K7 expression is a prognostic parameter in resected early stage NSCLC that allows identification of high-risk NSCLC patients with poor cancer-specific and overall survival.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cohort Studies; Female; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Reproducibility of Results; Retrospective Studies; Survival Rate

To date, a number of potential biomarkers for lung squamous cell cancer (SCC) have been identified; however, sensitive biomarkers are currently lacking to detect early stage SCC due to low sensitivity and specificity. In the present study, we compared the 7 serum proteomic profiles of 11 SCC patients, 7 chronic obstructive pulmonary disease (COPD) patients and 7 healthy smokers as controls to identify potential serum biomarkers associated with SCC and COPD. Two-dimensional difference gel electrophoresis (2D-DIGE) and mass-spectrometric analysis (MS) using an affinity column revealed two candidate proteins, haptoglobin (HP) and apolipoprotein 4, as biomarkers of SCC, and α-1-antichymotrypsin as a marker of COPD. The iTRAQ technique was also used to identify SCC-specific peptides. HP protein expression was significantly higher in SCC patients than in COPD patients. Furthermore, two HP protein peptides showed significantly higher serum levels in SCC patients than in COPD patients. We established novel polyclonal antibodies for the two HP peptides and subsequently a sandwich enzyme-linked immunosorbent assay (ELISA) for the quantification of these specific peptides in patient and control sera. The sensitivity of detection by ELISA of one HP peptide (HP216) was 70% of SCC patients, 40% of COPDs patients and 13% of healthy controls. We also measured CYFRA, a cytokeratin fragment clinically used as an SCC tumor marker, in all the 28 cases and found CYFRA was detected in only seven SCC cases. However, when the measurement of HP216 was combined with that of CYFRA, 100% (10 of 10 patients) of SCC cases were detected. Our proteomic profiling demonstrates that the SCC-specific HP peptide HP216 may potentially be used as a diagnostic biomarker for SCC.

Topics: Adult; Aged; alpha 1-Antichymotrypsin; Apolipoproteins A; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Electrophoresis, Gel, Two-Dimensional; Enzyme-Linked Immunosorbent Assay; Haptoglobins; Humans; Keratins; Lung Neoplasms; Male; Mass Spectrometry; Middle Aged; Peptide Fragments; Proteome; Pulmonary Disease, Chronic Obstructive; Smoking

Surgery is the treatment of choice for patients with non-small cell lung cancer (NSCLC) stages I-IIIA. However, more than 20% of these patients develop recurrence and die due to their disease. The release of tumor cells into peripheral blood (CTCs) is one of the main causes of recurrence of cancer. The objectives of this study are to identify the prognostic value of the presence and characterization of CTCs in peripheral blood in patients undergoing radical resection for NSCLC.. 56 patients who underwent radical surgery for previously untreated NSCLC were enrolled in this prospective study. Peripheral blood samples for CTC analysis were obtained before and one month after surgery. In addition CTCs were phenotypically characterized by epidermal growth factor receptor (EGFR) expression.. 51.8% of the patients evaluated were positive with the presence of CTCs at baseline. A decrease in the detection rate of CTCs was observed in these patients one month after surgery (32.1%) (p = 0.035). The mean number of CTCs was 3.16 per 10 ml (range 0-84) preoperatively and 0.66 (range 0-3) in postoperative determination. EGFR expression was found in 89.7% of the patients at baseline and in 38.9% patients one month after surgery. The presence of CTCs after surgery was significantly associated with early recurrence (p = 0.018) and a shorter disease free survival (DFS) (p = .008). In multivariate analysis CTC presence after surgery (HR = 5.750, 95% CI: 1.50-21.946, p = 0.010) and N status (HR = 0.296, 95% CI: 0.091-0.961, p = 0.043) were independent prognostic factors for DFS.. CTCs can be detected and characterized in patients undergoing radical resection for non-small cell lung cancer. Their presence might be used to identify patients with increased risk of early recurrence.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cohort Studies; ErbB Receptors; Female; Humans; Keratins; Longitudinal Studies; Lung Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplastic Cells, Circulating; Prognosis; Prospective Studies

To investigate the relationship between the presence of circulating tumor cells (CTC) and the values of coagulation factors including D-dimer (D-D), fibrinogen (FIB) and platelet (PLT) in primary lung cancer patients.. Peripheral venous blood samples were collected from 79 patients with previously untreated primary lung cancer. The levels of D-D, FIB and PLT were detected. The CTCs were enriched by negative immunomagnetic separation with anti-CD45 antibody and then detected by immunocytochemistry with Anti-pan Cytokeratin antibody. The relationship between these parameters and clinicopathologic characteristics of the patients was analyzed.. The levels of D-D, FIB and PLT were( 1.74±2.04) mg/L, (3.51±1.46 )g/L, (311±139)×10(9)/L, respectively. The level of D-D was associated with distant metastasis of lung cancer (P=0.046). The level of FIB was associated with clinical stage and distant metastasis (P<0.05). The level of PLT was associated with age, clinical stage and distant metastasis (all P<0.05). Among the 79 patients, there were 45 CTC-positive and 34 CTC-negative cases. The levels of D-D in the CTC-positive and CTC-negative groups were (2.31±2.41)mg/L and (0.99±1.02)mg/L, those of FIB were (3.79±1.56)g/L and (3.14±1.25)g/L, and those of PLT were (338±130)×10(9)/L and (229±129)×10(9)/L, respectively(P<0.05 for all). The positive rate of CTC was significantly higher in the metastasis group (82.8%), significantly higher than that in the non-metastatic group (42.0%, P<0.001). The levels of D-D, FIB and PLT in the metastasis group were (2.33±1.95)mg/L, (4.13±1.43)g/L and (433±74)×10(9)/L, but were (1.40±2.03) mg/L, (3.15±1.37)g/L and (206±88)×10(9)/L in the non-metastatic group (P<0.05 for all).. The detection of circulating tumor cells may facilitate early prediction of distant metastasis of lung cancer. The hypercoagulation state is more-likely correlated with the distant metastasis of lung cancer.

Topics: Age Factors; Blood Platelets; Fibrin Fibrinogen Degradation Products; Fibrinogen; Humans; Keratins; Lung Neoplasms; Neoplastic Cells, Circulating

Pleural effusion (PE), excess fluid in the pleural space, is often observed in lung cancer patients and also forms due to many benign ailments. Classifying it quickly is critical, but this remains an analytical challenge often lengthening the diagnosis process or exposing patients to unnecessary risky invasive procedures. We tested the analysis of PE using a multiplexed cytokeratin (CK) panel with targeted mass spectrometry-based quantitation for its rapid classification. CK markers are often assessed in pathological examinations for cancer diagnosis and guiding treatment course. We developed methods to simultaneously quantify 33 CKs in PE using peptide standards for increased analytical specificity and a simple CK enrichment method to detect their low amounts. Analyzing 121 PEs associated with a variety of lung cancers and noncancerous causes, we show that abundance levels of 10 CKs can be related to PE etiology. CK-6, CK-7, CK-8, CK-18, and CK-19 were found at significantly higher levels in cancer-related PEs. Additionally, elevated levels of vimentin and actin differentiated PEs associated with bacterial infections. A classifier algorithm effectively grouped PEs into cancer-related or benign PEs with 81% sensitivity and 79% specificity. A set of undiagnosed PEs showed that our method has potential to shorten PE diagnosis time. For the first time, we show that a cancer-relevant panel of simple-epithelial CK markers currently used in clinical assessment can also be quantitated in PEs. Additionally, while requiring less invasive sampling, our methodology demonstrated a significant ability to identify cancer-related PEs in clinical samples and thus could improve patient care in the future.

Topics: Actins; Aged; Biomarkers, Tumor; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Lung Neoplasms; Male; Mass Spectrometry; Middle Aged; Pleural Effusion; Vimentin

Polyhydroquinone-graphene composite as a new redox species was synthesized simply by a microwave-assisted one-pot method through oxidative polymerization of hydroquinone by graphene oxide, which exhibited excellent electrochemical redox activity at 0.124 V and can remarkably promote electron transfer. The as-prepared composite was used as immunosensing substrate in a label-free electrochemical immunosensor for the detection of cytokeratins antigen 21-1, a kind of biomarker of lung cancer. The proposed immunosensor showed wide liner range from 10 pg mL(-1) to 200 ng mL(-1) with a detection limit 2.3 pg mL(-1), and displayed a good stability and selectivity. In addition, this method has been used for the analysis of human serum sample, and the detection results showed good consistence with those of ELISA. The present substrate can be easily extended to other polymer-based nanocomposites.

Topics: Antigens; Biomarkers, Tumor; Composite Resins; Electrochemical Techniques; Graphite; Humans; Immunoassay; Keratins; Lung Neoplasms; Oxidation-Reduction; Quinones

Small cell lung cancer (SCLC) is characterized by prevalent circulating tumour cells (CTCs), early metastasis and poor prognosis. We show that SCLC patients (37/38) have rare CTC subpopulations co-expressing vascular endothelial-cadherin (VE-cadherin) and cytokeratins consistent with vasculogenic mimicry (VM), a process whereby tumour cells form 'endothelial-like' vessels. Single-cell genomic analysis reveals characteristic SCLC genomic changes in both VE-cadherin-positive and -negative CTCs. Higher levels of VM are associated with worse overall survival in 41 limited-stage patients' biopsies (P<0.025). VM vessels are also observed in 9/10 CTC patient-derived explants (CDX), where molecular analysis of fractionated VE-cadherin-positive cells uncovered copy-number alterations and mutated TP53, confirming human tumour origin. VE-cadherin is required for VM in NCI-H446 SCLC xenografts, where VM decreases tumour latency and, despite increased cisplatin intra-tumour delivery, decreases cisplatin efficacy. The functional significance of VM in SCLC suggests VM regulation may provide new targets for therapeutic intervention.

Topics: Animals; Antigens, CD; Biopsy; Cadherins; Cell Line, Tumor; Cohort Studies; DNA Copy Number Variations; Female; Humans; Keratins; Lung; Lung Neoplasms; Male; Mice; Middle Aged; Mutation; Neoplastic Cells, Circulating; Neovascularization, Pathologic; Single-Cell Analysis; Small Cell Lung Carcinoma; Tumor Suppressor Protein p53; Xenograft Model Antitumor Assays

A 7-year-old Afghan hound presented with a history of disorientation, loss of vision, and seizures. Magnetic resonance imaging helped identify a mass at the level of the main olfactory bulb that compressed and displaced adjacent tissues in the cribriform plate into the nasal cavity and nasopharynx. Bony structures were osteolytic. After removing almost 80% of the mass, the tumor recurred a few months later. Due to severe respiratory distress and subsequent to an ultrasound diagnosis of a liver tumor, the dog was euthanized. In addition to the nasal mass, a single nodule in the liver and multiple nodules in the lung were present. All masses had similar cell morphology and were diagnosed as metastasizing esthesioneuroblastoma. The neoplastic cells expressed neuron-specific enolase and chromogranin A, and a few cells within the nasal mass were positive for cytokeratin. This is the first description of a canine esthesioneuroblastoma with distant metastases.

Topics: Animals; Cerebrum; Chromogranin A; Dog Diseases; Dogs; Esthesioneuroblastoma, Olfactory; Keratins; Liver; Liver Neoplasms; Lung; Lung Neoplasms; Male; Nasal Cavity; Neoplasm Recurrence, Local; Nose Neoplasms; Phosphopyruvate Hydratase

Many tumor markers have been analyzed for applications in diagnosis, prognosis, and monitoring of cancer. Currently chemotherapy is routinely performed for patients with non-small cell lung cancer (NSCLC). The purpose of this study was to examine the serum tumor biomarker of cytokeratin (CK)-3A9 level in patients with NSCLC and its potential correlation with chemotherapeutic response.. The serum samples of 196 NSCLC patients, 84 healthy controls, and 87 benign lung disease patients were provided for measurement of CK-3A9 and carcinoembryonic antigen (CEA). Serum CK-3A9 concentration was examined using a chemoluminescent method. The potential correlation between serum CK18-3A9 concentration and chemotherapeutic response was analyzed in 124 patients with advanced NSCLC (stages III and IV).. The serum CK-3A9 levels in NSCLC patients pre-chemotherapy were significantly higher than those of healthy controls and benign lung disease patients (p < 0.01). CK-3A9 was related to Union for International Cancer Control (UICC) stages (p < 0.01) and histological classification (p < 0.05), but not related to age, gender, smoking status, and chemotherapy regimen (all p > 0.05). The testing results of serum CK-3A9 levels showed a higher sensitivity than that for CEA (48.2% and 39.5%, respectively). The chemotherapeutic response in the 124 patients with advanced NSCLC included 0 complete response (CR), 50 partial response (PR), 65 no change (NC), and 9 progression disease (PD). Post-chemotherapy CK-3A9 levels were significantly decreased compared to pre-chemotherapy (p < 0.05). The serum CK-3A9 levels in patients who achieved PR declined significantly compared to those who did not respond (SD + PD) after 2 cycles chemotherapy (p < 0.05).. CK-3A9 appeared to be a new biomarker for reliable, cost-effective prediction of the efficacy of chemotherapy in patients with advanced NSCLC, although the results should be confirmed in larger studies.

Topics: Adult; Aged; Antineoplastic Agents; Biomarkers; Carcinoma, Non-Small-Cell Lung; Case-Control Studies; Cisplatin; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged

EpCAM expressing circulating tumor cells, detected by CellSearch, are predictive of short survival in several cancers and may serve as a liquid biopsy to guide therapy. Here we investigate the presence of EpCAM(+) CTC detected by CellSearch and EpCAM(-) CTC discarded by CellSearch, after EpCAM based enrichment. EpCAM(-) CTC were identified by filtration and fluorescent labelling. This approach was validated using different cell lines spiked into blood and evaluated on blood samples of 27 metastatic lung cancer patients. The majority of spiked EpCAM(+) cells could be detected with CellSearch, whereas most spiked cells with EpCAM(low) or EpCAM(-) expression were detected using filtration. Five or more CTC were detected in 15% of the patient samples, this increased to 41% when adding the CTC detected in the discarded blood. The number of patients with CTC and the number of CTC detected were doubled by the presence of EpCAM(-) CTC. In this pilot study, the presence of EpCAM(+) CTC was associated with poor outcome, whereas the EpCAM(-) CTC were not. This observation will need to be confirmed in larger studies and molecular characterization needs to be conducted to elucidate differences between EpCAM(-) and EpCAM(+) CTC.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Case-Control Studies; Cell Adhesion Molecules; Cell Line, Tumor; Epithelial Cell Adhesion Molecule; Female; Flow Cytometry; Follow-Up Studies; Humans; Kaplan-Meier Estimate; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Neoplastic Cells, Circulating; Prognosis

Peritoneal mesothelioma is rare, and the sarcomatoid variant is more infrequent, with <30 cases reported to date in the literature. Several case series have described the morphologic features of sarcomatoid peritoneal mesothelioma (SPe); however, the clinicopathologic features are not well characterized. To our knowledge, this is the first large series reporting the clinicopathologic features of SPe. We reviewed our database of 3106 malignant mesothelioma cases. Of 248 peritoneal mesotheliomas, 15 (4%) were sarcomatoid variant (0.5% of all mesotheliomas). Only cases with 100% sarcomatoid morphology diagnosed by open surgical biopsy and/or autopsy were included. Thus, 4 cases were excluded leaving 11 cases of SPe. Two additional cases of SPe previously published by 1 of the authors (V.L.R.), not included in the database, are added yielding 13 cases total. The median age at diagnosis was 66 years (range=48 to 85 y), and there was a male predominance (M:F=3.25:1). Survival from date of diagnosis to date of death was 5 months (range=0 to 12 mo). The most common presenting symptom was abdominal pain, and 3 of 4 women were suspected to have cholecystitis/cholelithiasis. All cases stained positive for cytokeratins, and 2 contained heterologous elements. Seven cases had objective markers of asbestos exposure, and 2 additional cases had occupations strongly associated with mesothelioma. Two cases with alleged household contact exposures could not be confirmed to be asbestos related by lung fiber analysis. SPe is a rare variant of mesothelioma attributed to asbestos exposure in 69% of our cases.

Topics: Aged; Aged, 80 and over; Asbestos; Autopsy; Biomarkers, Tumor; Biopsy; Databases, Factual; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Incidence; Inhalation Exposure; Keratins; Lung Neoplasms; Male; Mesothelioma; Mesothelioma, Malignant; Middle Aged; Occupational Exposure; Peritoneal Neoplasms; Predictive Value of Tests; Prognosis; Risk Factors; Sarcoma; Survival Analysis

Four cases of tumors in which cell internalization was frequently visualized are reported: one feline mammary carcinoma, one feline cutaneous squamous cell carcinoma, one canine pulmonary squamous cell carcinoma and one canine pleural mesothelioma. Cell internalization was observed by cytology in two of these cases (the feline mammary tumour and the pleural effusion in the canine mesothelioma) and by histopathology in all but the canine mesothelioma. Immunohistochemical staining for pancytokeratin was positive for both internalized and host cells, while E-cadherin expression was frequently absent, although internalized cells occasionally stained positive. This cell-to-cell interaction seems to be associated with tumors displaying a strong epithelial-mesenchymal transitional phenotype, in which cancer cells become engulfed by other cancer cells. Such event could be regarded as an important hallmark of very high malignancy.

Topics: Animals; Biomarkers, Tumor; Biopsy; Cadherins; Carcinoma, Squamous Cell; Cat Diseases; Cats; Cytophagocytosis; Dog Diseases; Dogs; Female; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Mammary Neoplasms, Animal; Mesothelioma; Pleural Neoplasms; Skin Neoplasms

Topics: Carcinoid Tumor; Diagnosis, Differential; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Paraganglioma

The enumeration and characterization of circulating tumor cells (CTCs), found in the peripheral blood of cancer patients, provide a potentially accessible source for cancer diagnosis and prognosis. This work reports on a novel spiral microfluidic device with a trapezoidal cross-section for ultra-fast, label-free enrichment of CTCs from clinically relevant blood volumes. The technique utilizes the inherent Dean vortex flows present in curvilinear microchannels under continuous flow, along with inertial lift forces which focus larger CTCs against the inner wall. Using a trapezoidal cross-section as opposed to a traditional rectangular cross-section, the position of the Dean vortex core can be altered to achieve separation. Smaller hematologic components are trapped in the Dean vortices skewed towards the outer channel walls and eventually removed at the outer outlet, while the larger CTCs equilibrate near the inner channel wall and are collected from the inner outlet. By using a single spiral microchannel with one inlet and two outlets, we have successfully isolated and recovered more than 80% of the tested cancer cell line cells (MCF-7, T24 and MDA-MB-231) spiked in 7.5 mL of blood within 8 min with extremely high purity (400-680 WBCs mL(-1); ~4 log depletion of WBCs). Putative CTCs were detected and isolated from 100% of the patient samples (n = 10) with advanced stage metastatic breast and lung cancer using standard biomarkers (CK, CD45 and DAPI) with the frequencies ranging from 3-125 CTCs mL(-1). We expect this simple and elegant approach can surmount the shortcomings of traditional affinity-based CTC isolation techniques as well as enable fundamental studies on CTCs to guide treatment and enhance patient care.

Topics: Antibodies, Immobilized; Breast Neoplasms; Cell Line, Tumor; Cell Separation; Cell Survival; Female; Fluorescein-5-isothiocyanate; Humans; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Microfluidic Analytical Techniques; Microscopy, Confocal; Neoplastic Cells, Circulating; Receptor, ErbB-2

Detection and characterization of circulating tumor cells (CTCs) may reveal insights into the diagnosis and treatment of malignant disease. Technologies for isolating CTCs developed thus far suffer from one or more limitations, such as low throughput, inability to release captured cells, and reliance on expensive instrumentation for enrichment or subsequent characterization. We report a continuing development of a magnetic separation device, the magnetic sifter, which is a miniature microfluidic chip with a dense array of magnetic pores. It offers high efficiency capture of tumor cells, labeled with magnetic nanoparticles, from whole blood with high throughput and efficient release of captured cells. For subsequent characterization of CTCs, an assay, using a protein chip with giant magnetoresistive nanosensors, has been implemented for mutational analysis of CTCs enriched with the magnetic sifter. The use of these magnetic technologies, which are separate devices, may lead the way to routine preparation and characterization of "liquid biopsies" from cancer patients.

Topics: Antibodies, Immobilized; Antigens, Neoplasm; Carcinoma, Non-Small-Cell Lung; Cell Adhesion Molecules; Cell Line, Tumor; Cell Separation; Epithelial Cell Adhesion Molecule; ErbB Receptors; Fluorescein-5-isothiocyanate; Humans; Keratins; Lung Neoplasms; Magnetics; Magnetite Nanoparticles; MCF-7 Cells; Microfluidic Analytical Techniques; Mutation; Neoplastic Cells, Circulating

Ewing's sarcoma/primitive neuroectodermal tumor (PNET) has been the subject of recent reports describing morphologic variants (adamantinoma-like, large cell, spindle cell, sclerosing, clear cell, and vascular-like) of the most classic form, as well as cases displaying unusual morphologic differentiation and atypical immunohistochemical features. We report a case of an uncommon lung tumor in a 20-year-old female, morphologically and molecularly consistent with an Ewing's sarcoma/PNET tumor with foci of squamous differentiation, and peculiar expression of vimentin, high-molecular-weight keratins, p63, synaptophysin, and chromogranin. This case raises a challenging differential diagnostic problem with therapeutic implications: Should the patient be treated following the protocols for Ewing's sarcoma/PNET tumors or as for lung carcinoma with neuroendocrine features? The patient we report here was treated with neoadjuvant chemotherapy for Ewing's sarcoma/PNET according to Euro Ewing 99 study protocol followed by surgery and has no evidence of disease 15 months after the initial diagnosis. This highlights the importance of achieving the correct diagnosis of these atypical tumors using all clinical, morphological, and ancillary methods available to allow for their correct and timely treatment.

Topics: Antigens, CD; Biomarkers, Tumor; Female; Humans; Keratins; Lung Neoplasms; Neuroectodermal Tumors, Primitive; Vimentin; Young Adult

Immunohistochemical (IHC) staining for the identification of nodal occult metastases (OM), not detected by routine histological examination, has been proposed for improved staging, prognostication and decision of adjuvant treatment in surgically treated primary lung cancer. In a prospective study, we analysed 178 cases of primary lung cancer stage I-III (N0-N1) for OM by immunostaining lymph node tissue using a broad-spectrum anti-cytokeratin antibody. OM were found in 7 (4 %) of the 178 cases. Using Kaplan-Meier analysis, overall survival was not significantly different between cases with stage I and cases upstaged to stage II because of OM (n = 3), or between cases with stage II and cases upstaged to stage III (n = 4). Likewise, the presence of OM was not significantly correlated with overall survival in univariable or multivariable Cox proportional hazards regression models, also when disregarding OM <0.2 mm in size. Given the low frequency of OM and lack of significant impact on survival in our study, the justification for including IHC staining of lymph nodes in lung cancer in clinical practise does not appear convincing. Moreover, we report several potential pitfalls in the use of broad-spectrum cytokeratin IHC staining for OM detection, for example staining of intra-nodal mesothelial cells.

Topics: Humans; Immunohistochemistry; Kaplan-Meier Estimate; Keratins; Lung Neoplasms; Lymphatic Metastasis; Neoplasm Staging; Prospective Studies; Staining and Labeling; Sweden

Cervical cancer is the seventh most common malignancy in both genders combined and the third most common cancer in women. Despite significant progress in treatments, cervical cancer is not completely curable. Therefore, further research is necessary in this area. Animal models are one of the most practical tools in the field of cancer research. The present study aimed to characterize the growth behavior and surface markers of HeLa cells after heterotopic and systemic inoculation to athymic nude mice.. Ten 6-week old female athymic C57BL/6 nude mice were used in this study. HeLa cells were inoculated into the flank or tail vein. The tumor volume was calculated and growth curves were drawn. Tumor-bearing mice were sacrificed and the lesions obtained after harvesting were analyzed in a pathology lab. Subsequently, one slide per tumor was stained with hematoxylin and eosin (H&E) and other slides were stained immunohistochemically by cytokeratins (CK), vimentin, P53, CD34, and Ki-67.. Tumor take rate, mean doubling time and latency period were 94.4%, 5.29 ± 3.57 days and 15.27 days, respectively. H&E results revealed highly malignant hyperchromatin epithelial cells. Immunohistochemical examination of the heterotopic tumors indicated greater expression of CK and less expression of vimentin compared to the metastatic ones. Sixty percent of cells were P53-positive and more than 80% were Ki-67-positive. CD34 expression indicated the intensity of angiogenesis in tumor.. This study represents a comprehensive description of a HeLa xenograft model for in vivo investigations, enabling researchers to assess new treatments for cervical cancer.

Topics: Animals; Antigens, CD34; Carcinoma; Disease Models, Animal; Female; HeLa Cells; Heterografts; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Lung Neoplasms; Mice; Mice, Inbred C57BL; Mice, Nude; Time Factors; Tumor Burden; Tumor Suppressor Protein p53; Uterine Cervical Neoplasms; Vimentin

Patients with small cell lung carcinoma (SCLC) rarely demonstrate long-term survival. We previously reported that gene expression profiling identified a subset of SCLC with good prognosis in surgical cases. To find an easier way to routinely identify SCLC belonging to this subset, we conducted the present study with a hypothesis that neuroendocrine (NE) or basaloid (BA) phenotypes may influence prognosis. To confirm the subset, we used an array platform to analyze fresh samples. Because inoperable cases may differ from surgical cases, we enrolled 51 biopsy cases and 43 resected samples. To evaluate NE and BA phenotypes, we used NE (synaptophysin, chromogranin A, and CD56) and BA (p63 and CK34βE12) markers. To varying extents, expression profiling based on the array platform duplicated the subsets. For NE phenotypes, 77% of surgical cases and 100% of biopsy cases were positive for at least 1 marker. For BA phenotypes, only 19% of surgical cases were positive for at least 1 marker, whereas there were no positive biopsy cases. Cases undergoing surgery were categorized based on NE and BA immunoreactivity; 58% into NE+BA-, 19% into NE+BA+, 23% into NE-BA-, and 0 into NE-BA+ groups. NE- patients (n = 10) demonstrated a significantly better prognosis (P = .0306) than their NE+ counterparts (n = 33), whereas no survival difference was evident between the BA+ and BA- groups. Multivariate analyses showed that NE positivity was an independent prognostic factor. In conclusion, the SCLC subset with good prognosis is identified by low NE marker expression, which was found only in surgical cases.

Topics: Aged; Biomarkers, Tumor; Biopsy; Carcinoma, Neuroendocrine; CD56 Antigen; Chromogranin A; Female; Gene Expression Profiling; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Membrane Proteins; Middle Aged; Prognosis; Protein Array Analysis; Small Cell Lung Carcinoma; Synaptophysin

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Calbindin 2; Cholecystitis; Cisplatin; Cystitis; Diagnosis, Differential; Female; Glutamates; Guanine; Humans; Inflammation; Keratins; Lung Neoplasms; Male; Mesothelioma; Mesothelioma, Malignant; Middle Aged; Pemetrexed; Peritoneal Neoplasms; Survival Rate; Vimentin

This study aimed at evaluating the utility of a panel of antibodies, consisting of cytokeratins (CK) 5/6, CK34βE12, p63, thyroid transcription factor-1 (TTF-1), and CK7 for distinguishing between squamous cell carcinoma (SCC) and adenocarcinomas (Ad), as well as their expression with clinicopathological parameters and prognosis in SCC and Ad. 111 SCC of small biopsy specimens and 99 cases of Ad were stained by immunohistochemistry, among which 76 SCC and 64 Ad had complete follow-up data. Most of SCC displayed CK5/6 (91/99, 91.92%), CK34βE12 (83/99, 83.84%), p63 (96/99, 96.97%), and most of Ad showed expression of CK7 (89/111, 80.18%) and TTF-1 (105/111, 94.59%). The combination of CK5/6/CK34βE12/p63 seems to be useful for differentiating SCC from Ad with 100% sensitivity and 97.30% specificity, and TTF-1 was a useful biomarker for Ad with 94.59% sensitivity and 100% specificity. There were differences between CK5/6, p63, and TTF-1 expression with tumor differentiation (p<0.05) in SCC or Ad. Univariate analysis indicated that patients with high TTF-1 expression predicted better prognosis in Ad patients. Multivariate analysis showed that TTF-1 expression (HR=0.340, 95% CI 0.143-0.811, p=0.015) was a good independent predictor of Ad patient survival.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Biopsy; Diagnosis, Differential; DNA-Binding Proteins; Female; Humans; Immunohistochemistry; Keratin-5; Keratin-6; Keratin-7; Keratins; Lung Neoplasms; Male; Membrane Proteins; Middle Aged; Transcription Factors; Young Adult

Circulating tumor microemboli (CTM) are potentially important cancer biomarkers, but using them for cancer detection in early-stage disease has been assay limited. We examined CTM test performance using a sensitive detection platform to identify stage I non-small-cell lung cancer (NSCLC) patients undergoing imaging evaluation.. First, we prospectively enrolled patients during 18F-FDG PET-CT imaging evaluation for lung cancer that underwent routine phlebotomy where CTM and circulating tumor cells (CTCs) were identified in blood using nuclear (DAPI), cytokeratin (CK), and CD45 immune-fluorescent antibodies followed by morphologic identification. Second, CTM and CTC data were integrated with patient (age, gender, smoking, and cancer history) and imaging (tumor diameter, location in lung, and maximum standard uptake value [SUVmax]) data to develop and test multiple logistic regression models using a case-control design in a training and test cohort followed by cross-validation in the entire group.. We examined 104 patients with NSCLC, and the subgroup of 80 with stage I disease, and compared them to 25 patients with benign disease. Clinical and imaging data alone were moderately discriminating for all comers (Area under the Curve [AUC] = 0.77) and by stage I disease only (AUC = 0.77). However, the presence of CTM combined with clinical and imaging data was significantly discriminating for diagnostic accuracy in all NSCLC patients (AUC = 0.88, p value = 0.001) and for stage I patients alone (AUC = 0.87, p value = 0.002).. CTM may add utility for lung cancer diagnosis during imaging evaluation using a sensitive detection platform.

Topics: Aged; Aged, 80 and over; Area Under Curve; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Embolism; Female; Fluorodeoxyglucose F18; Humans; Indoles; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Male; Middle Aged; Multimodal Imaging; Neoplasm Staging; Neoplastic Cells, Circulating; Positron-Emission Tomography; Prospective Studies; Radiopharmaceuticals; Risk Assessment; Tomography, X-Ray Computed; Tumor Burden

Presence and frequency of rare circulating tumor cells (CTCs) in bloodstreams of cancer patients are pivotal to early cancer detection and treatment monitoring. Here, we use a spiral microchannel with inherent centrifugal forces for continuous, size-based separation of CTCs from blood (Dean Flow Fractionation (DFF)) which facilitates easy coupling with conventional downstream biological assays. Device performance was optimized using cancer cell lines (> 85% recovery), followed by clinical validation with positive CTCs enumeration in all samples from patients with metastatic lung cancer (n = 20; 5-88 CTCs per mL). The presence of CD133⁺ cells, a phenotypic marker characteristic of stem-like behavior in lung cancer cells was also identified in the isolated subpopulation of CTCs. The spiral biochip identifies and addresses key challenges of the next generation CTCs isolation assay including antibody independent isolation, high sensitivity and throughput (3 mL/hr); and single-step retrieval of viable CTCs.

Topics: AC133 Antigen; Antigens, CD; Blood Cells; Cell Line, Tumor; Cell Separation; Centrifugation; Glycoproteins; Humans; Keratins; Leukocyte Common Antigens; Lung Neoplasms; MCF-7 Cells; Microfluidic Analytical Techniques; Neoplastic Cells, Circulating; Peptides

The epithelial-mesenchymal transition (EMT) is a program involved in embryonic development that is often activated during cancer invasion and metastasis. CD133 is the main marker identifying cancer stem cells (CSCs) in lung cancer. Circulating tumor cells (CTCs) are demonstrated to be useful as a biomarker for the diagnosis and treatment of cancer. The aim of this study was to correlate EMT, CSCs and CTCs with patient prognosis to verify whether they can contribute to better stratification of lung cancer patients at risk for recurrent and metastatic disease. Pulmonary venous blood was drawn after major pulmonary surgery in 45 patients with resectable non-small cell lung cancer (NSCLC) in order to identify CTCs. For the same patients, we also constructed prognostic lung tissue microarrays (TMA) for CD133 and c-kit and evaluated CSC and EMT markers using flow cytometry. Cytokeratin-positive cells were detectable in 11 (23.9%) cases. c-kit expression was heterogeneous in prognostic TMAs while CD133 expression was low or absent which was also confirmed by flow cytometry and RT-PCR. Flow cytometric analysis showed that the mean percentage of cells with CD133 expression was 1.6%. CD90 and CD326 markers were co-expressed with a mean percentage of 10.41%. When CD133 and CD90/CD326 expression was correlated with follow-up, CD133 showed a higher correlation with deceased patients when compared with CD90/CD326 co-expression (32.5 vs. 9.5%). CD133 expression demonstrated a strong significant association with patients exhibiting progressive disease when compared to CD90/CD326 expression (15 vs. 7.1%). CD133 may be significantly associated with invasion and metastatic spread of NSCLC. The co-expression of CD90, CD326 and CD133 has definite prognostic value in patients with NSCLC.

Topics: Antigens, CD; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Epithelial-Mesenchymal Transition; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplastic Cells, Circulating; Neoplastic Stem Cells; Prognosis; Proto-Oncogene Proteins c-kit; Pulmonary Veins

Small-cell lung cancer (SCLC) has a very aggressive clinical course with early metastasis. This study investigated how the distinctive neuroendocrine characteristics contribute to disease progression and invasion in human SCLC.. The neuroendocrine phenotype (pro-opiomelanocortin (POMC)) was quantified by ELISA in blood samples from 43 SCLC patients. The neuroendocrine (POMC, chromogranin A, neuron-specific enolase, NCAM) and epithelial (cytokeratin and E-cadherin) phenotypes were investigated, using ELISA and immunocytochemistry/immunohistochemistry.. In SCLC patients, 16% had elevated circulating POMC, which was associated with significantly worse survival (P=0.02) and liver metastases (P=0.004). In addition, POMC correlated with epithelial-positive circulating tumour cells (P=0.0002). In a panel of SCLC cell lines, all POMC-secreting cell lines expressed cytokeratin (40% of total). Even after cloning, DMS 79 cells expressed both neuroendocrine and epithelial markers. DMS 79 xenografts secreted POMC into the blood, which mirrored the tumour volume. These xenografts expressed both neuroendocrine and epithelial phenotypes in all tumours, with both phenotypes prevalent in cells invading the surrounding tissue.. Both neuroendocrine and epithelial phenotypes coexist in human SCLC tumours in vitro and in vivo and this persists in invading tumour cells. In patients, POMC secretion predicts poor survival and liver metastases, suggesting a crucial role of the neuroendocrine phenotype.

Topics: Animals; Cadherins; Carcinoma, Small Cell; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Humans; Keratins; Liver Neoplasms; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Metastasis; Neoplastic Cells, Circulating; Neuroendocrine Cells; Phenotype; Pro-Opiomelanocortin; Survival Rate; Transplantation, Heterologous

The use of p63 has been advocated for separating small cell lung carcinoma from poorly differentiated non-small cell lung carcinoma, in particular, squamous cell lung carcinoma. However, p63 is not entirely specific in this distinction, as several studies have demonstrated p63 immunoreactivity in a proportion of small cell lung carcinomas. p40 (ΔNp63) has recently been purported to be a highly specific marker for squamous cell lung carcinoma, but data regarding p40 (ΔNp63) in small cell lung carcinoma, a key differential diagnostic consideration in biopsy samples of squamous cell lung carcinoma, are lacking. In this study, a total of 34 previously confirmed small cell lung carcinomas (27 bronchoscopic biopsy samples and 7 large specimens) were immunostained for p40 (ΔNp63), p63, and keratin 34βE12. All 34 small cell lung carcinomas were negative for p40 (ΔNp63) and keratin 34βE12. Although none of the large small cell lung carcinoma specimens exhibited p63 immunoreactivity, 12 (44.4%) of 27 biopsy samples of small cell lung carcinoma were positive for p63. The rate of p63 staining in small cell lung carcinoma biopsy samples differed significantly from that of p40 (ΔNp63) and keratin 34βE12 (P = .005). Ten cases of squamous cell lung carcinoma were also tested, all of which were positive for all 3 markers. These findings confirm that p63 immunoexpression is not uncommon in biopsy samples of small cell lung carcinoma. Positive p63 staining may be mistakenly interpreted as supportive of squamous differentiation and result in misclassification of small cell lung carcinoma as squamous cell lung carcinoma, a diagnostic error that has important therapeutic and prognostic consequences. To provide greater diagnostic accuracy, p40 (ΔNp63) or keratin 34βE12 should be used instead of p63 in the distinction of small cell lung carcinoma from non-small cell lung carcinoma in biopsy samples.

Topics: Biomarkers, Tumor; Biopsy; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Small Cell Lung Carcinoma

Epithelial-to-mesenchymal transition (EMT), the phenotypical change of cells from an epithelial to a mesenchymal type, is thought to be a key event in invasion and metastasis of adenocarcinomas. These changes involve loss of keratin expression as well as loss of cell polarity and adhesion. We here aimed to determine whether the loss of keratin expression itself drives increased invasion and metastasis in adenocarcinomas and whether keratin loss leads to the phenotypic changes associated with EMT. Therefore, we employed a recently described murine model in which conditional deletion of the Keratin cluster II by Cre-recombinase leads to the loss of the entire keratinmultiprotein family. These mice were crossed into a newly generated Cre-recombinase inducible KRAS-driven murine lung cancer model to examine the effect of keratin loss on morphology, invasion and metastasis as well as expression of EMT related genes in the resulting tumors. We here clearly show that loss of a functional keratin cytoskeleton did not significantly alter tumor morphology or biology in terms of invasion, metastasis, proliferation or tumor burden and did not lead to induction of EMT. Further, tumor cells did not induce synchronously expression of vimentin, which is often seen in EMT, to compensate for keratin loss. In summary, our data suggest that changes in cell shape and migration that underlie EMT are dependent on changes in signaling pathways that cause secondary changes in keratin expression and organization. Thus, we conclude that loss of the keratin cytoskeleton per se is not sufficient to causally drive EMT in this tumor model.

Topics: Animals; Biomarkers; Carcinoma, Non-Small-Cell Lung; Cell Line; Disease Models, Animal; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Gene Order; Gene Targeting; Genes, ras; Humans; Keratins; Lung Neoplasms; Mice; Mice, Transgenic; Small Cell Lung Carcinoma

The diagnostic test for ALK rearrangement in non-small-cell lung cancer (NSCLC) for crizotinib treatment is currently done on tumor biopsies or fine-needle aspirations. We evaluated whether ALK rearrangement diagnosis could be performed by using circulating tumor cells (CTCs).. The presence of an ALK rearrangement was examined in CTCs of 18 ALK-positive and 14 ALK-negative patients by using a filtration enrichment technique and filter-adapted fluorescent in situ hybridization (FA-FISH), a FISH method optimized for filters. ALK-rearrangement patterns were determined in CTCs and compared with those present in tumor biopsies. ALK-rearranged CTCs and tumor specimens were characterized for epithelial (cytokeratins, E-cadherin) and mesenchymal (vimentin, N-cadherin) marker expression. ALK-rearranged CTCs were monitored in five patients treated with crizotinib.. All ALK-positive patients had four or more ALK-rearranged CTCs per 1 mL of blood (median, nine CTCs per 1 mL; range, four to 34 CTCs per 1 mL). No or only one ALK-rearranged CTC (median, one per 1 mL; range, zero to one per 1 mL) was detected in ALK-negative patients. ALK-rearranged CTCs harbored a unique (3'5') split pattern, and heterogeneous patterns (3'5', only 3') of splits were present in tumors. ALK-rearranged CTCs expressed a mesenchymal phenotype contrasting with heterogeneous epithelial and mesenchymal marker expressions in tumors. Variations in ALK-rearranged CTC levels were detected in patients being treated with crizotinib.. ALK rearrangement can be detected in CTCs of patients with ALK-positive NSCLC by using a filtration technique and FA-FISH, enabling both diagnostic testing and monitoring of crizotinib treatment. Our results suggest that CTCs harboring a unique ALK rearrangement and mesenchymal phenotype may arise from clonal selection of tumor cells that have acquired the potential to drive metastatic progression of ALK-positive NSCLC.

Topics: Adult; Aged; Anaplastic Lymphoma Kinase; Cadherins; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Crizotinib; Female; Gene Rearrangement; HeLa Cells; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratins; Lung Neoplasms; Male; MCF-7 Cells; Middle Aged; Neoplastic Cells, Circulating; Protein Kinase Inhibitors; Pyrazoles; Pyridines; Receptor Protein-Tyrosine Kinases; Vimentin

Epithelial-mesenchymal transition (EMT) is defined as switching of polarized epithelial cells to a migratory fibroblastoid phenotype. EMT is known to be involved in the progression and metastasis of various cancers. The aim was to evaluate that whether EMT-related proteins' alterations are associated with clinicopathological features and prognosis in lung adenocarcinoma.. The expression of EMT-related proteins including cytokeratin, E-cadherin, TTF-1, β-catenin, vimentin, Snail, Twist, CD44 was evaluated by immunohistochemistry using a tissue array method in the lung adenocarcinoma tissues of 95 patients. In addition, clinicopathological characteristics and survival were compared with the expression of EMT-related proteins.. Loss of epithelial proteins and/or acquisition of the expression of mesenchymal proteins were observed in lung adenocarcinoma. These proteins' alteration was associated with poor cell differentiation and poor patients' outcome, respectively. Subjects were divided into two groups according to the number of EMT-related proteins' alteration. A higher number of EMT-related proteins' alteration was found to be significantly associated with unfavorable outcome. Multivariate analysis showed that a higher number of EMT-related proteins' alteration was independently associated with poor prognosis.. The number of EMT-related proteins' alteration is a significant prognostic marker to predict overall survival in patients with lung adenocarcinoma. The information generated will be valuable for the prognosis of patients with lung adenocarcinoma.. The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1007838329872974.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Adult; Aged; beta Catenin; Cadherins; Epithelial-Mesenchymal Transition; Female; Gene Expression Regulation, Neoplastic; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Prognosis; Thyroid Nuclear Factor 1; Transcription Factors; Vimentin

When adenocarcinomas arise within the esophagus, particularly when located away from the gastroesophageal junction, it may be important in some patients to differentiate between a primary esophageal adenocarcinoma and metastasis from another site. Lung adenocarcinoma is one tumor that has been reported to frequently metastasize to the esophagus.. To create a panel of immunohistochemical markers that can reliably distinguish between an esophageal and pulmonary primary; within the gastrointestinal pathology literature, including published articles and textbooks, common lung immunohistochemical markers, such as TTF-1, are assumed to be negative in esophageal adenocarcinoma, yet, to our knowledge, no study has yet investigated the veracity of that presumption.. In this study, 24 cases each of pulmonary and esophageal adenocarcinomas were stained with TTF-1, napsin A, CDX2, 34βE12, N-cadherin, and IMP3 in an attempt to define an optimal panel for differentiation. Esophageal adenocarcinomas occurring at the gastroesophageal junction were excluded in this study because a gastric primary tumor cannot be excluded in those cases.. Surprisingly, TTF-1 and napsin A were positive in similar proportions of tumors from both sites. Those markers that differentiated statistically between esophageal and pulmonary adenocarcinoma were IMP3, CDX2, and N-cadherin.. When differentiating the origin of a tumor as either esophageal or pulmonary, an immunohistochemical panel consisting of IMP3, CDX2, and N-cadherin is superior to either TTF-1 or napsin A.

Topics: Adenocarcinoma; Antigens, CD; Aspartic Acid Endopeptidases; Biomarkers, Tumor; Cadherins; CDX2 Transcription Factor; Diagnosis, Differential; Esophageal Neoplasms; Homeodomain Proteins; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Nuclear Proteins; RNA-Binding Proteins; Staining and Labeling; Thyroid Nuclear Factor 1; Transcription Factors

Early-stage non-small cell lung cancer (NSCLC) patients have a high risk of disease relapse despite curatively intended surgical resection, and the detection of tumour cells in the bone marrow could be one method of determining the presence of the disseminated disease in its early stages.. Bone marrow aspirates were collected from 296 patients at the time of surgery, and the presence of disseminated tumour cells was determined with the help of immunomagnetic selection (IMS) using the MOC31-antibody recognising EpCAM and with the help of standard immunocytochemistry (ICC) using the anti-cytokeratin (CK) antibodies AE1/AE3.. Disseminated tumour cells were found in 152 of 252 (59%) bone marrow samples using IMS and in 25 of 234 (11%) samples using ICC. No association between the two detection methods was observed. The presence of EpCAM⁺ cells was not associated with any clinicopathological parameters, whereas a higher frequency of CK⁺ cells was found in patients with an advanced pT status. Disseminated tumour cells, as detected using IMS, had no prognostic impact. Patients with CK⁺ cells in the bone marrow had a reduced relapse-free survival, but the difference was not statistically significant.. Our findings do not support the further development of DTC detection for clinical use in early-stage NSCLC. Future studies should include the molecular characterisation of DTCs, along with an attempt to identify subpopulations of cells with biological and clinical significance.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Antigens, Neoplasm; Bone Marrow Cells; Carcinoma, Non-Small-Cell Lung; Cell Adhesion Molecules; Epithelial Cell Adhesion Molecule; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Neoplasm Recurrence, Local; Neoplastic Cells, Circulating; Treatment Outcome

Recent advances in genome wide transcriptional analysis have provided greater insights into the etiology and heterogeneity of breast cancer. Molecular signatures have been developed that stratify the conventional estrogen receptor positive or negative categories into subtypes that are associated with differing clinical outcomes. It is thought that the expression patterns of the molecular subtypes primarily reflect cell-of-origin or tumor driver mutations. In this study however, using a genetically engineered mouse mammary tumor model we demonstrate that the PAM50 subtype signature of tumors driven by a common oncogenic event can be significantly influenced by the genetic background on which the tumor arises. These results have important implications for interpretation of "snapshot" expression profiles, as well as suggesting that incorporation of genetic background effects may allow investigation into phenotypes not initially anticipated in individual mouse models of cancer.

Topics: Animals; Animals, Outbred Strains; Female; Genetic Association Studies; Genetic Loci; Genetic Predisposition to Disease; Haplotypes; Humans; Keratins; Lung Neoplasms; Male; Mammary Neoplasms, Experimental; Mice; Mice, Transgenic; Transcriptome

Topics: Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Biopsy; Bone Neoplasms; Carcinoma, Small Cell; Chromogranin A; Fatal Outcome; Head and Neck Neoplasms; Humans; Keratins; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Prostate-Specific Antigen; Prostatic Neoplasms; Scalp; Skin Neoplasms; Tomography, X-Ray Computed

Basaloid squamous cell carcinoma is a rare and aggressive variant of squamous cell carcinoma, which mostly occurs in the upper aerodigestive tracts. Basaloid squamous cell carcinoma also typically arises in the anal canal, but is extremely rare in the lower gastrointestinal tract. A 70-year-old man presented with loose stool and intermittent hematochezia 2 months ago. Colonoscopy showed an ulceroinfiltrative mass on the rectosigmoid colon from 16 cm to 18 cm above the anal verge. Conventional colonoscope could not pass through the lesion but it was possible with pediatric colonoscope. Abdominal CT scan showed 1.6 cm sized wall thickening with circumferential luminal narrowing in the rectosigmoid colon and multiple ill-defined low density masses in both lobes of the liver. Therefore, colon cancer with liver metastasis was suspected. However, basaloid cells were noted on histologic examination, and they were weakly positive for synaptophysin on immunohistochemical study. After palliative lower anterior resection, histologic examination of the resected specimen revealed basaloid differentiation with keratin pearls, and tumor cells were positively stained with high molecular weighted cytokeratin (34BE12) and CK 5/6. Thus, the patient was finally diagnosed with basaloid squamous cell carcinoma of rectosigmoid colon with distant metastases.

Topics: Aged; Carcinoma, Squamous Cell; Colonoscopy; Colorectal Neoplasms; Humans; Immunohistochemistry; Keratins; Liver Neoplasms; Lung Neoplasms; Male; Positron-Emission Tomography; Synaptophysin; Tomography, X-Ray Computed

Bone marrow-derived stem cells (BMDCs) have the ability to differentiate into lung epithelial cells in response to damage; however, their role in squamous cell carcinoma (SCC) formation is unknown. This study aimed to determine whether BMDC-derived lung epithelial cells could contribute to SCC formation. A model of lung SCC induced with N-nitrosodiethylamine (NDEA) in recipient female mice transplanted with green fluorescent protein (GFP)-positive BMDCs from male donors was established. Incorporation of BMDCs in lung tissue was determined using immunohistochemistry and immunofluorescence to detect GFP expression and fluorescence in situ hybridization to Y chromosomes. BMDC appeared at three stages of lung SCC progression: metaplasia, dysplasia, and carcinoma. There was a significantly higher proportion of GFP-positive (GFP(+)) cells within SCC than was found in metaplasia and dysplasia 16 weeks post-transplantation (both P < 0.017); GFP(+) BMDCs were also observed in clusters within several SCC nests. Furthermore, most GFP(+) cells in SCC were pancytokeratin-positive (PCK(+)) epithelial cells, and some exhibited proliferative activity as determined by Ki67 staining (9.7 ± 3.92 %). The presence of GFP(+)Ki67(+)PCK(+) cells within SCC nests suggested that some donor BMDCs differentiated into proliferating epithelial cells. Finally, analysis of p63 expression, a marker of SCC cells, indicated that the presence of GFP(+)p63(+) cells (green) in inner parts of the SCC. These findings strongly suggest that BMDC-derived lung epithelial cells could participate in lung SCC formation and partially contribute to tumor growth, which might have significant potential implications for both clinical cancer therapy using BMDCs.

Topics: Animals; Bone Marrow Cells; Bone Marrow Transplantation; Carcinoma, Squamous Cell; Cell Differentiation; Cell Movement; Cell Proliferation; Cell Transformation, Neoplastic; Diethylnitrosamine; Epithelial Cells; Female; Green Fluorescent Proteins; Keratinocytes; Keratins; Ki-67 Antigen; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Phosphoproteins; Stem Cells; Trans-Activators

Single cell analysis and cell sorting has enabled the study of development, growth, differentiation, repair and maintenance of "liquid" tissues and their cancers. The application of these methods to solid tissues is equally promising, but several unique technical challenges must be addressed. This report illustrates the application of multidimensional flow cytometry to the identification of candidate stem/progenitor populations in non-small cell lung cancer and paired normal lung tissue. Seventeen paired tumor/normal lung samples were collected at the time of surgical excision and processed immediately. Tissues were mechanically and enzymatically dissociated into single cell suspension and stained with a panel of antibodies used for negative gating (CD45, CD14, CD33, glycophorin A), identification of epithelial cells (intracellular cytokeratin), and detection of stem/progenitor markers (CD44, CD90, CD117, CD133). DAPI was added to measure DNA content. Formalin fixed paraffin embedded tissue samples were stained with key markers (cytokeratin, CD117, DAPI) for immunofluorescent tissue localization of populations detected by flow cytometry. Disaggregated tumor and lung preparations contained a high proportion of events that would interfere with analysis, were they not eliminated by logical gating. We demonstrate how inclusion of doublets, events with hypodiploid DNA, and cytokeratin+ events also staining for hematopoietic markers reduces the ability to quantify epithelial cells and their precursors. Using the lung cancer/normal lung data set, we present an approach to multidimensional data analysis that consists of artifact removal, identification of classes of cells to be studied further (classifiers) and the measurement of outcome variables on these cell classes. The results of bivariate analysis show a striking similarity between the expression of stem/progenitor markers on lung tumor and adjacent tumor-free lung.

Topics: Adenocarcinoma; Biomarkers; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; DNA; DNA, Neoplasm; Epithelial Cells; Flow Cytometry; Humans; Keratins; Lung; Lung Neoplasms; Proto-Oncogene Proteins c-kit; Stem Cells

It is difficult to distinguish desmoplastic malignant mesothelioma (DMM) from fibrous pleuritis (FP). We investigated the utility of immunohistochemistry as a way of differentiating between DMM and FP. We examined 11 DMMs and 46 FPs with the aid of antibodies against 18 cytokeratin (CK) subtypes, calponin, caldesmon, desmin, and GLUT-1. The best sensitivity and specificity cut-off values in the receiver operating characteristic curves (ROC) for CKs 7, 8, 17, 18, and 19, and GLUT-1 were each above 60%. When cases with either DMM or FP were partitioned by the staining score associated with the best sensitivity and specificity cut-off values in ROC, the incidence of a positive expression for CKs 7, 8, 17, 18, and 19, and GLUT-1 was significantly higher in DMM than in FP. In conclusion, immunohistochemistry for CKs 7, 8, 17, 18, and 19, and GLUT-1 may be useful, alongside histological characteristics, for separating DMM from FP.

Topics: Aged; Aged, 80 and over; Biomarkers; Biomarkers, Tumor; Diagnosis, Differential; Female; Glucose Transporter Type 1; Humans; Immunohistochemistry; Keratin-17; Keratin-18; Keratin-19; Keratin-7; Keratin-8; Keratins; Lung Neoplasms; Male; Mesothelioma; Mesothelioma, Malignant; Middle Aged; Pleurisy; Retrospective Studies

The use of supervised classification to extract markers from primary flow cytometry data is an emerging field that has made significant progress, spurred by the growing complexity of multidimensional flow cytometry. Whether the markers are extracted without supervision or by conventional gate and region methods, the number of candidate variables identified is typically larger than the number of specimens (p > n) and many variables are highly intercorrelated. Thus, comparison across groups or treatments to determine which markers are significant is challenging. Here, we utilized a data set in which 86 variables were created by conventional manual analysis of individual listmode data files, and compared the application of five multivariate classification methods to discern subtle differences between the stem/progenitor content of 35 nonsmall cell lung cancer and adjacent normal lung specimens. The methods compared include elastic-net, lasso, random forest, diagonal linear discriminant analysis, and best single variable (best-1). We described a broadly applicable methodology consisting of: 1) variable transformation and standardization; 2) visualization and assessment of correlation between variables; 3) selection of significant variables and modeling; and 4) characterization of the quality and stability of the model. The analysis yielded both validating results (tumors are aneuploid and have higher light scatter properties than normal lung), as well as leads that require followup: Cytokeratin+ CD133+ progenitors are present in normal lung but reduced in lung cancer; diploid (or pseudo-diploid) CD117+CD44+ cells are more prevalent in tumor. We anticipate that the methods described here will be broadly applicable to a variety of multidimensional cytometry problems.

Topics: AC133 Antigen; Adenocarcinoma; Antigens, CD; Biomarkers; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Epithelial Cells; Flow Cytometry; Glycoproteins; Humans; Hyaluronan Receptors; Keratins; Lung; Lung Neoplasms; Peptides; Proto-Oncogene Proteins c-kit; Sensitivity and Specificity; Stem Cells

Primary lymphoepithelioma-like carcinoma (LELC) of the lung is an extremely rare disease that occurs more commonly in Asians, and is composed of undifferentiated carcinoma with prominent lymphoid stroma. LELC is reported to be closely associated with Epstein-Barr virus (EBV) infection. A case is presented here in which bronchial brushing smears in a 70-year-old man, revealed large clusters of neoplastic cells with scant cytoplasm. The nuclei were large, hyperchromatic, of irregular contour and with prominent nucleoli. Also identified were prominent intratumoral lymphoid infiltration and brisk mitotic figures. We detected EBV-coded small RNA in situ hybridization in smears. A cytologic diagnosis of a LELC was suggested. Further evaluation and immunohistochemical studies were conducted on formalin-fixed, paraffin-embedded material. Cords or nests of large neoplastic cells with enlarged nuclei and prominent nucleoli with marked lymphoid infiltration and lymphoid stroma were identified on H&E sections. Immunohistochemically, the tumor cells showed diffuse and strong membranous staining for CK(AE1/AE3), CK5/6, CK34βE12, Napsin A and Bcl-2 but were negative for CK7, CK14, CK20, EMA, TTF-1, chromogranin A, synaptophysin and CD56. The proliferative index with MIB-1 was around 60%, and the p53 positive cells around 20%. The diagnosis of primary LELC of the lung was confirmed based on cytopathologic, histopathologic, immunohistochemical and EBER results, and a detailed systemic examination to exclude possible extrapulmonary (nasopharyngeal) origin. We report the cytopathological features of LELC of the lung and demonstrate here for the first time the positivity of the EBER with RNA-ISH method in smears with emphasis on differential diagnostic considerations.

Topics: Aged; Aspartic Acid Endopeptidases; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Nucleus; Cell Proliferation; Cytoplasm; Diagnosis, Differential; Herpesvirus 4, Human; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Lung Neoplasms; Male; RNA, Viral

Lung cancer is the main cause of cancer-related death in Western countries. Despite early diagnosis, approximately 40% of patients have undergone surgical resection for localized non-small cell lung cancer relapse within 24 months after surgery. Current prognostic criteria for patients with non-small cell lung cancer are gradually enriched by the discovery of critical biological markers in surgical samples to better stratify patients with high risk for recurrent and metastatic disease after surgical manipulation. In fact, specific biological features are needed to drive metastasis development and, among these chemokine receptors, when activated, seem to play a relevant role, promoting both neovessels formation and tumoral cell migration.. To this purpose, blood samples from the closed stumps of the pulmonary veins were drawn immediately after major pulmonary surgery in 45 patients with resectable non-small cell lung cancer to evaluate the expression of chemokine CXCL12 receptor, CXCR4, in circulating tumor cells. In addition, primary tumor sections have been used to assess microvascular density (MVD) and vessels invasion and build prognostic tissue micro-array to investigate the expression of CXCL12 receptors CXCR4 and CXCR7.. Cells positive for cytokeratins from tumor draining pulmonary venous blood were detectable in 11 cases (23.9%). In 8 out of 11 cases, CK positive cells coexpressed CXCR4. Moreover, in tumoral tissue high CXCR4 expression was significantly associated to high mMVD (p = 0.046), high CXCR7 expression (p = 0.001), adenocarcinoma histotype (p = 0.023), and to the presence of circulating tumoral cells in pulmonary veins (p = 0.001). Finally, vessel invasions relate to high MVD.. In conclusion, the results of our study underline the significant potential role of CXCL12 receptors in determining both vessel formation and tumoral cell migration to blood stream, favoring metastasis development.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Chemokine CXCL12; Epidemiologic Methods; Female; Humans; Keratins; Lung Neoplasms; Male; Microvessels; Middle Aged; Neoplasm Invasiveness; Neoplasm Proteins; Neoplasm Staging; Neoplastic Cells, Circulating; Prognosis; Receptors, CXCR; Receptors, CXCR4

Proper evaluation of lung nodules is a difficult issue for clinical management of patients. Discriminating metastatic endometrial stromal sarcoma (ESS) from other primary spindle cell neoplasms of the lung using histological analysis can be challenging. This is particularly true when an adequate clinical history is lacking, because ESS metastasis can be delayed by a couple of decades. To emphasize the importance of the correlation of pathological findings with clinical history and imaging studies, we investigated 11 cases of ESS (seven low grade and four high grade) metastatic to the lung. All cases presented with one to multiple unilateral or bilateral lung nodules that were detected by chest computed tomography. Primary ESS was diagnosed from hysterectomy specimens except for one by endometrial biopsy, 0.5 to 23 years prior to metastasis. Immunohistochemical studies showed that all ESS cases were moderately to strongly positive for Bcl-2 and CD10 with >50% of tumor cells stained, except for one high grade ESS that was negative for CD10. Eight (72.7%) and seven (63.6) of the 11 cases showed positive estrogen and progesterone receptors, respectively, with a majority of positive cases showing diffuse and moderate to strong staining. Strong but patchy staining for CD34 was detected in one (9.1%) case with smooth muscle differentiation. CK7 and TTF-1 were negative in all cases. Two (18.2%) cases exhibited patchy and strong positivity for caldesmon. Two (18.2%) low grade ESS cases showed moderate to strong AE1/AE3 positivity in >50% of tumor cells, one of which also showed moderate CK19 and Cam 5.2 staining in >30% of tumor cells. One should be cautious when assessing spindle cell neoplasms of the lung in women with a history of hysterectomy. Correlation of clinical history and imaging studies with histological and immunohistochemical findings is essential to diagnosis of metastatic ESS to the lung.

Topics: Aged; Biomarkers, Tumor; Diagnosis, Differential; Endometrial Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Neprilysin; Proto-Oncogene Proteins c-bcl-2; Radiography; Sarcoma, Endometrial Stromal; Staining and Labeling

Epithelial-myoepithelial carcinoma (EMC) is a rare low-grade salivary gland malignancy of presumed intercalated duct origin comprising 1% of all salivary gland tumours. High grade transformation (HGT) in EMC is a recently recognised entity with only a few cases reported in the literature. The authors report an additional case of EMC with HGT involving the submandibular gland. The patient was a 60-year-old woman who requested examination of the rapid growth of a mass in the left submandibular area, which she had first noticed 20 years previously. Histologically, the tumour had two distinct carcinomatous components. One component had features of a low grade EMC. The second component consisted of polygonal cells, arranged in a solid and nested pattern, with marked nuclear pleomorphism, brisk mitotic activity, and frequent necrosis. The Ki-67 labelling index of the EMC component was 9%, and that of the high grade component was 40%. The patient developed multiple pulmonary metastases 15 months after surgery. The aggressive behaviour of EMC with HGT suggests that it is important to recognise this variant of EMC to avoid misdiagnosis and inappropriate treatment.

Topics: Calcium-Binding Proteins; Calmodulin-Binding Proteins; Calponins; Carcinoma; Cell Nucleus; Cell Transformation, Neoplastic; Cytoplasm; Diagnostic Errors; Female; Follow-Up Studies; Humans; Keratins; Ki-67 Antigen; Lung Neoplasms; Membrane Proteins; Microfilament Proteins; Middle Aged; Mitotic Index; Necrosis; Neoplasm Invasiveness; Submandibular Gland Neoplasms

In this study, we report 2 pediatric cases of nuclear protein of the testis (NUT) midline carcinoma (NMC) suggestive of pulmonary origin: case 1 was a 14-year-old Japanese boy and case 2 was a 7-year-old Japanese girl. Initial symptoms of both cases were prolonged cough and chest pain, and the case 2 patient also complained of lumbago and lumbar mass due to bone metastases. Imaging studies revealed that pulmonary tumors from both patients were located at the hilar region of the lower lobe. Biopsies of the tumors showed undifferentiated carcinoma in case 1 and combined undifferentiated and squamous cell carcinoma in case 2. Despite intensive treatment with chemotherapy and radiation, progression of neither tumor was controlled, and both patients died of the tumors at 1 year (case 1) and 4 months (case 2) after onset of disease. Both tumors were diffusely positive for p63 and NUT expression and were partially positive for various cytokeratins. Reverse transcription polymerase chain reaction analysis and subsequent direct sequencing revealed that the bromodomain-containing protein 4-NUT chimeric gene was present in tumor tissue of both patients, leading to a diagnosis of NMC. The tumor cells of case 1 were also positive for thyroid transcription factor-1 expression, but those of case 2 were negative. Histologic examination of the surgically removed lung tumor of case 1 indicated that the origin of the tumor was basal cells of the bronchiolar epithelia.

Topics: Adolescent; Biomarkers, Tumor; Biopsy; Carcinoma, Squamous Cell; Cell Differentiation; Child; Disease Progression; Fatal Outcome; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Neoplasm Proteins; Nuclear Proteins; Oncogene Proteins; Oncogene Proteins, Fusion; Reverse Transcriptase Polymerase Chain Reaction; Spinal Neoplasms; Thyroid Nuclear Factor 1; Time Factors; Tomography, X-Ray Computed; Transcription Factors; Treatment Outcome; Tumor Suppressor Proteins

Detection of micrometastases plays an important role in early-stage and recurrent cancer diagnosis. In the study, a new method of screening micrometastases of lung cancer in peripheral blood by magnetic nanoparticles (MNPs) and quantum dots (QDs) was developed to achieve early diagnosis and recurrence prevention. MNPs were prepared by combining miniemulsion polymerization and Stöber coating methods. QDs were prepared by using Cd(Ac)(2) · 2H(2)O and oxygen-free NaHTe with thioglycolic acid as the stabilizer. The carbodiimide-mediated condensation method was used to couple pan-cytokeratin (pan-ck) antibody (Ab) to the surface of the MNPs, and Lunx and SP-A Abs to the surface of the QDs. After four kinds of epithelial tumor cells were enriched by MNPs coupled with pan-ck Ab (MNP-pan-ck), lung cancer cells A549 and SPC-A-1 were successfully identified by QDs with double-labeled Abs. Finally, 32 patients with non-small cell lung cancer (NSCLC) were collected, out of 26 cases with the enriched circulating tumor cells (CTCs), 21 cases were successfully identified by QDs. Therefore, a new method was established in which MNP-pan-ck collected CTCs and QDs with double-labeled Abs could be used simultaneously to identify CTCs from NSCLC patients.

Topics: Aged; Cadmium Compounds; Carcinoma, Non-Small-Cell Lung; Case-Control Studies; Cell Line, Tumor; Female; Glycoproteins; Hep G2 Cells; Humans; Immunohistochemistry; Keratins; Leukocytes, Mononuclear; Lung Neoplasms; Magnetite Nanoparticles; Male; Middle Aged; Neoplasm Micrometastasis; Neoplastic Cells, Circulating; Phosphoproteins; Pulmonary Surfactant-Associated Protein A; Quantum Dots; Tellurium

To establish and characterize a lung adenocarcinoma cell line from a female patient in Xuanwei, Yunnan province.. Surgical specimen of the lung adenocarcinoma was obtained and cultured immediately in RPMI 1640 medium with 10% fetal bovine serum and 10(5) U/L penicillin and 100 mg/L streptomycin. When stable proliferation of the cells was achieved after over 40 passages in culture, the biological features of the cell line were investigated by cell morphology, karyotyping, protein marker expression [cytokeratins (CKs), epithelial membrane antigen (EMA) and CD proteins], growth kinetics, cell cycle phase distribution, mitotic index, colony formation in soft agar, cell invasion and tumorigenicity in Balb/c nude mice.. The established cell line was stably cultured for over 80 passages during a one-year period as an anchorage-dependent monolayer of short spindle, polygonal to epithelioid cells under phase contrast microscope. Microglandular cavities and disordered microfilaments were observed under transmission electron microscope. The growth curve presented in an "S" shape with the cell population doubled every 46.7 hours. The mitotic index was 1.5% and the colony formation rate was 8.3%. The cell cycle distribution included 76.9% in G(0)/G(1), 15.1% in S and 8.0% in G(2)/M. The cell line displayed a hypotriploid karyotype with a mode of 66 chromosomes and a median of 64 chromosomes. The cells expressed CK7, CK8, CK (Pan) and EMA by immunohistochemistry. A high level of cell surface expression of CD13 and CD59 was evident by flow cytometry. The cells were able to penetrate Matrigel in vitro but failed to form a stable xenograft in nude mice.. A new human lung adenocarcinoma cell line, designated as XLA-07, is successfully established from a Xuanwei lung cancer patient.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Animals; CD13 Antigens; CD59 Antigens; Cell Culture Techniques; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Female; Humans; Karyotyping; Keratins; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Mucin-1; Neoplasm Transplantation; Polyploidy; Tumor Stem Cell Assay

A 6-year-old Girolando dairy cow was presented for evaluation of a large subcutaneous facial mass. Fine-needle aspirates of the mass contained many neoplastic cells with high nuclear:cytoplasmic ratios arranged in sheets and loosely cohesive clusters with streaming erythrocytes and neutrophils in the background. Neoplastic cells were 13-25 μm in diameter and were round to cuboidal with variably distinct borders. Based on the signalment, anatomic location, and cytologic findings, differential diagnoses included salivary adenocarcinoma, squamous cell carcinoma, and mucoepidermoid carcinoma. The cow was euthanized and a necropsy was performed. The primary neoplasm arose from the left parotid salivary gland and meastatic tumor was found in the regional lymph nodes and lung. Histologically, the tumor was composed of anastomosing and irregular solid islets surrounded by scant stroma. Cells were negative for periodic acid-Schiff (PAS), PAS-diastase, and Alcian blue pH 2.5 stains, used to detect mucin. On immunohistochemical analysis, neoplastic luminal salivary gland cells expressed cytokeratin, but not S100, α-smooth muscle actin, or vimentin. Peripheral cells of neoplastic islets were immunoreactive for p63. The final diagnosis was nonsecretory adenocarcinoma of the parotid salivary gland.

Topics: Adenocarcinoma; Animals; Biopsy, Fine-Needle; Cattle; Cattle Diseases; Diagnosis, Differential; Face; Fatal Outcome; Female; Keratins; Lung; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Parotid Gland; Parotid Neoplasms

Myelosarcomas are, due to their rarity, a difficult differential diagnosis. Not infrequently, extensive immunohistochemical staining for characterization of the tumor is performed, if one does not directly think of myelosarcoma. In the present case, there was a positivity of the myeloid blasts for cytokeratin. This may complicate the discrimination of myelosarcoma from carcinoma, in particular small cell carcinoma, not only in the mediastinum, but also in the skin, e.g., Merkel cell carcinoma.

Topics: Adenocarcinoma; Biomarkers, Tumor; Cell Transformation, Neoplastic; Diagnosis, Differential; Female; Humans; Keratins; Leukemia, Myeloid, Acute; Lung Neoplasms; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Mediastinum; Middle Aged; Myeloid Cells; Neoplasms, Multiple Primary

To study the clinicopathologic features and histologic differential diagnosis of small cell neuroendocrine carcinoma (SmCC) of kidney.. The clinicopathologic features of 12 cases of SmCC of kidney encountered during the period from 1999 to 2010 were retrospectively reviewed.. Six cases of primary and 6 cases of metastatic SmCC involving kidney were identified. Amongst the primary renal SmCC, 2 were located in renal parenchyma and 4 in renal pelvis. Chest X-ray showed negative findings. Five of them underwent radical nephrectomy. On gross examination, the tumor was located centrally around the renal pelvis in 4 cases and peripherally in renal parenchyma in 1 case. On the other hand, 4 of the 6 cases of metastatic SmCC were discovered during therapy for pulmonary SmCC. Two of these patients presented with abdominal pain and gross hematuria, with lung and renal tumor masses identified simultaneously. The diagnosis of all the 6 cases of metastatic SmCC was confirmed by fine needle aspiration biopsy. Microscopically, pure SmCC was demonstrated in the 2 cases of primary renal parenchymal SmCC and 6 cases of metastatic SmCC. The 4 primary renal pelvic SmCC coexisted with urothelial carcinoma component. On immunohistochemical study, all cases were positive for cytokeratin, synaptophysin and CD56. All metastatic cases and 4 primary cases were also positive for TTF-1. Of six patients with primary SmCC two died 4 and 9 months after operation, and two were alive with a follow-up of 25 and 138 months, respectively. Five of six cases with metastatic SmCC died 3 - 8 months after diagnosis. The other 3 cases were failed to follow-up.. Both primary and metastatic SmCC can be found in the kidney. Although rare, primary SmCC is located either in renal parenchyma or in pelvis. The diagnosis of SmCC relies on morphologic examination and immunohistochemical study. TTF-1 immunostaining cannot reliably distinguish primary from metastatic SmCC in kidney. Correlation with clinicoradiologic findings and demonstration of coexisting urothelial carcinoma component (if any) is helpful in delineation of the tumor origin.

Topics: Adult; Aged; Carcinoma, Neuroendocrine; Carcinoma, Renal Cell; Carcinoma, Small Cell; CD56 Antigen; Diagnosis, Differential; Female; Follow-Up Studies; Humans; Keratins; Kidney Neoplasms; Lung Neoplasms; Lymphoma; Male; Middle Aged; Nephrectomy; Nuclear Proteins; Retrospective Studies; Sarcoma, Ewing; Synaptophysin; Thyroid Nuclear Factor 1; Transcription Factors; Treatment Outcome; Wilms Tumor

Infundibulocystic basal cell carcinoma (IBCC) is a variant of basal cell carcinoma. Sporadic cases usually represent a solitary tumor and multiple IBCC is rare. There have been no reports in which the tumor differentiation is characterized immunohistochemically. We report a case of multiple IBCC which developed on a patient's scalp by performing histopathological and immunohistochemical examinations, using monoclonal antibodies against cytokeratins (CKs). A 76-year-old female had noticed multiple small papules on her scalp. She noticed that the tumors were growing when she underwent systemic chemotherapy for metastatic lung cancer. Routine histopathological specimens from skin biopsies revealed findings typical of IBCC. The tumor cells expressed CK14 and CK17. However, CK1 and CK10 were expressed only in a few cells in the inner area of the tumors. The present case is unique in two points. First, multiple tumors developed on the patient's scalp during the systemic chemotherapy for the lung cancer. Second, the tumor showed CK expression patterns characteristic to infundibular and trichilemmal epithelia.

Topics: Adenocarcinoma; Aged; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Camptothecin; Carboplatin; Carcinoma, Basal Cell; Docetaxel; Female; Humans; Immunohistochemistry; Irinotecan; Keratins; Lung Neoplasms; Neoplasms, Second Primary; Skin Neoplasms; Taxoids

Topics: Aged; Biomarkers, Tumor; Diagnosis, Differential; Fatal Outcome; Humans; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Lymphatic Metastasis; Neoplasm Proteins; Small Cell Lung Carcinoma

Immunohistochemistry is increasingly utilized to differentiate lung adenocarcinoma and squamous cell carcinoma. However, detailed analysis of coexpression profiles of commonly used markers in large series of whole-tissue sections is lacking. Furthermore, the optimal diagnostic algorithm, particularly the minimal-marker combination, is not firmly established. We therefore studied whole-tissue sections of resected adenocarcinoma and squamous cell carcinoma (n=315) with markers commonly used to identify adenocarcinoma (TTF-1) and squamous cell carcinoma (p63, CK5/6, 34βE12), and prospectively validated the devised algorithm in morphologically unclassifiable small biopsy/cytology specimens (n=38). Analysis of whole-tissue sections showed that squamous cell carcinoma had a highly consistent immunoprofile (TTF-1-negative and p63/CK5/6/34βE12-diffuse) with only rare variation. In contrast, adenocarcinoma showed significant immunoheterogenetity for all 'squamous markers' (p63 (32%), CK5/6 (18%), 34βE12 (82%)) and TTF-1 (89%). As a single marker, only diffuse TTF-1 was specific for adenocarcinoma whereas none of the 'squamous markers,' even if diffuse, were entirely specific for squamous cell carcinoma. In contrast, coexpression profiles of TTF-1/p63 had only minimal overlap between adenocarcinoma and squamous cell carcinoma, and there was no overlap if CK5/6 was added as a third marker. An algorithm was devised in which TTF-1/p63 were used as the first-line panel, and CK5/6 was added for rare indeterminate cases. Prospective validation of this algorithm in small specimens showed 100% accuracy of adenocarcinoma vs squamous cell carcinoma prediction as determined by subsequent resection. In conclusion, although reactivity for 'squamous markers' is common in lung adenocarcinoma, a two-marker panel of TTF-1/p63 is sufficient for subtyping of the majority of tumors as adenocarcinomas vs squamous cell carcinoma, and addition of CK5/6 is needed in only a small subset of cases. This simple algorithm achieves excellent accuracy in small specimens while conserving the tissue for potential predictive marker testing, which is now an essential consideration in advanced lung cancer specimens.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Algorithms; Biomarkers, Tumor; Biopsy; Carcinoma, Squamous Cell; Diagnosis, Differential; DNA-Binding Proteins; Female; Humans; Immunohistochemistry; Keratin-5; Keratin-6; Keratins; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Prospective Studies; Reproducibility of Results; Sensitivity and Specificity; Transcription Factors; Tumor Suppressor Proteins

Topics: Adult; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Confounding Factors, Epidemiologic; Diagnosis, Differential; Epithelium; Humans; Keratins; Lung Neoplasms; Lymph Nodes; Lymphadenitis; Lymphatic Metastasis; Sentinel Lymph Node Biopsy

Lung cancer cells express several cytokeratins (CKs) that are subdivided into type I (CK9-23) and type II (CK1-8) subclasses. The functions of CKs in lung cancer cells have not been fully elucidated. The purpose of this study was to investigate the role of CKs in the invasion of lung cancer cells. We investigated the expression levels of CK7, 8, 18 and 19 in 12 non-small cell lung cancer (NSCLC) and seven SCLC cell lines by quantitative immunoblotting. The expression levels of these four CKs were significantly higher in the NSCLC cells. The NSCLC cell line HI1017 expressed CK8 and 18; A549 cells expressed CK7, 8, 18 and 19, respectively. Invasive sublines of HI1017 and A549 were established by repeated selection of invasive cells using a membrane invasion chamber system. The invasive cell lines showed lower expression levels of CKs compared with the parental cells. Exogenous CK19 also resulted in a decrease in invasiveness of the HI1017 cells. Suppression of either CK8 or CK18 by short interfering RNAs led to a decrease in the total CKs and increased invasiveness of both the HI1017 and A549 cells. A549 cells expressed very low levels of CK19. Suppression of CK19 affected neither invasive ability nor total CK amount in the A549 cells. Our observations indicate that CK expression levels were inversely associated with invasiveness of the NSCLC cell lines, and suggest that expression levels of dominant CKs may affect invasive ability.

Topics: Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Humans; Immunoblotting; Keratins; Lung Neoplasms; Neoplasm Invasiveness; RNA Interference; Small Cell Lung Carcinoma

The expression patterns of cytokeratin (CK) filaments in human epithelial neoplasms are complex and distinctive. The aims of this study were to analyze CK expression and to evaluate the diagnostic application of CKs in human non-small cell lung cancer (NSCLC).. mRNA expression of CK5, CK6, CK14, CK15, CK17, and CK19 was analyzed by Northern blotting. Protein expression of CK5/6, CK7, CK14, CK17, and CK18 was evaluated by immunohistochemistry on tissue microarrays.. Northern blotting showed that CKs were highly expressed in human bronchial epithelial cells and/or small airway epithelial cells. In NSCLC cell lines, the expression pattern of CKs was heterogeneous. Regarding protein expression of CKs in 95 primary lung tumors, expression of CK5/6, CK14, and CK17 proteins was increased in squamous cell carcinomas compared to adenocarcinomas (ADC; p = 0.001, p = 0.030, and p = 0.001, respectively), and higher expression was significantly associated with lower grading (p = 0.006, p = 0.002, and p = 0.001, respectively), while increased expression of CK7 and CK18 was observed in ADC (p = 0.001, respectively).. Our data suggest that CK5/6, CK7, CK14, CK17, and CK18 have diagnostic value in the subclassification of NSCLC.

Topics: Adenocarcinoma; Biomarkers, Tumor; Blotting, Northern; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Cell Line, Tumor; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratin-14; Keratin-17; Keratin-18; Keratin-5; Keratin-6; Keratin-7; Keratins; Lung Neoplasms; Protein Array Analysis; RNA, Messenger

Metastatic pulmonary tumors secondary to breast cancer detected either before or after surgery are predominantly multiple and bilateral. However, in cases detected to have a solitary pulmonary nodule (SPN), determining whether the lesion represents a primary cancer, metastasis, or a benign pulmonary lesion can be difficult.. Between January 2000 and December 2009, we performed breast cancer surgery on 1,226 patients, of which 49 cases (3.9%) were detected to have pulmonary lesions before or after the surgery. In 14 of these patients, video-assisted thoracoscopic surgery was performed to remove a SPN.. Pathological examination of the resected specimens in these 14 cases revealed metastatic pulmonary tumor in 8 cases, primary lung cancer in 3 cases, and benign disease in 3 cases. While lobectomy was performed in one of these patients with metastatic pulmonary tumor, the remaining 7 underwent partial resection of the lung. The primary lung cancer was an adenocarcinoma in all 3 patients, and lobectomy plus mediastinal lymph node dissection was performed in these patients. The tumor grading based on pathological diagnosis was T1N0M0, p-Stage 1A in all 3 patients. The prognosis was good in the breast cancer patients in whom the metastatic lung tumor was a SPN.. Evaluating the immunohistochemical cytokeratin profile and levels of the TTF-1 and GCDFP-15 of the lesion was useful when distinguishing between pulmonary cancer and metastatic pulmonary tumor. In addition, some patients exhibited changes in the biological properties of the metastatic tumor, and delete tumor resection by video-assisted thoracoscopic surgery can be useful for deciding the drug treatment strategy in some cases.

Topics: Adenocarcinoma; Adult; Aged; Breast Neoplasms; Female; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Lymph Node Excision; Mastectomy; Middle Aged; Neoplasm Grading; Prognosis; Solitary Pulmonary Nodule; Tomography, X-Ray Computed

Topics: Adenocarcinoma, Papillary; Adult; Diagnosis, Differential; Female; Follow-Up Studies; Humans; Keratin-7; Keratins; Lung Neoplasms; Nasopharyngeal Neoplasms; Nuclear Proteins; Thyroid Neoplasms; Thyroid Nuclear Factor 1; Transcription Factors

We investigated the staining pattern of commonly used basal cell/myoepithelial markers, such as p63 (a p53-homologous nuclear protein), basal cell-specific cytokeratin antibody (34betaE12, K903), and smooth muscle myosin heavy chain (SMMHC) in benign and malignant bronchioloalveolar proliferations of the lung. We studied 85 lung lesions consisting of 35 bronchioloalveolar carcinoma, 30 well-differentiated adenocarcinoma, and 20 cases of benign lung lesions. In normal lung, p63, K903, and SMMHC decorated the basal cells of large and small airways and occasional cells of terminal bronchioles. In reactive processes, a distinctive staining pattern was present in 19/20 (95%) of the cases characterized by staining of basal cells of the airways and bronchiolar epithelium and squamous metaplastic epithelium for p63 and K903, whereas 12/20 (60%) stained with SMMHC. Respiratory ciliated cells, alveolar epithelial cells, and nonepithelial cells were negative. In bronchioloalveolar carcinoma, a discontinuous peripheral rim of p63-immunoreactive cells was retained surrounding and intermingled with the malignant bronchioloalveolar proliferation in 31/35 (88.5%) cases, SMMHC in 28/35 (80%) cases, and K903 in 20/35 (57%) cases. For adenocarcinoma, a majority of the cases (28/30, 93%) were negative for p63 and K903; however, SMMHC showed artifactual staining in the desmoplastic stroma in 6/30 (20%) cases. Our results highlighted the differential expression of basal cell markers across various bronchioloalveolar lesions. The staining pattern of basal cells in bronchioloalveolar carcinoma supports that these neoplasms may actually be carcinoma in-situ.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Basal Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Membrane Proteins; Middle Aged; Predictive Value of Tests; Prognosis; Sensitivity and Specificity; Smooth Muscle Myosins

Primary renal synovial sarcoma is rare and might be misdiagnosed as another renal tumor. This study demonstrates the clinicopathologic and immunohistochemical features, differential diagnosis, and prognosis of such tumors.. Histologic slides and clinical data were reviewed for 4 patients with primary renal synovial sarcoma and immunohistochemical staining was performed. Molecular analysis was performed on 2 cases to demonstrate the presence of the SYT-SSX gene fusion transcripts by reverse transcriptase polymerase chain reaction (RT-PCR).. The patients were 2 women and 2 men aged from 32 to 48 years. The tumors were 10.0-15.0 cm in diameter, grey-white and solid, and hemorrhage or necrosis was observed. Microscopically, the tumors consisted of mitotically active, monomorphic plump spindle cells with indistinct cell borders growing in short, intersecting fascicles. Hypocellular myxoid areas and a prominent hemangiopericytomatous pattern were present in all cases. The average mitotic rate was 5-8 mitoses/10 high-power fields. Hemorrhage and tumor necrosis were easily found. Scattered small cysts lined with flat, cuboidal, or hobnailed epithelia were found in 3 cases. Tumor cells are immunoreactive for Vimentin (4/4), Bcl-2 (4/4), CD99 (4/4), and CD56 (3/4), and focally for EMA (3/4) and Cytokeratin (3/4). SYT-SSX1 gene fusion was detected in the 2 cases in which RT-PCR analysis was performed. One patient had tumor metastasis to the lung 6 months after surgery and died 5 months later. Multiple metastasis to the liver occurred in one patient and the patient died 13 months after the initial surgery. The other 2 patients had tumors recur at 8 and 15 months and died at 18 and 21 months, respectively, after the initial operation.. Primary renal synovial sarcoma is rare, with poor prognosis, characterized by SYT-SSX gene fusion, and needs to be differentiated from other renal sarcomas.

Topics: 12E7 Antigen; Adult; Antigens, CD; CD56 Antigen; Cell Adhesion Molecules; Female; Follow-Up Studies; Humans; Keratins; Kidney Neoplasms; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Mucin-1; Neoplasm Recurrence, Local; Nephrectomy; Oncogene Proteins, Fusion; Proto-Oncogene Proteins c-bcl-2; Sarcoma, Synovial; Survival Rate; Vimentin

We report an unusual case of hepatocellular carcinoma with three histologically and immunohistochemically distinct components, arising in a noncirrhotic liver, with a pulmonary tumor embolus from one of the three components. The histological patterns of the three components were fibrolamellar, well-differentiated nodular, and pleomorphic. The immunophenotypes were, respectively CK7- /CK20- /Hep Par1+, CK7+ /CK20- /Hep Par1+, and CK7+ /CK20+ /Hep Par1-. The tumor in the pulmonary embolus showed only the morphological and immunohistochemical features of the pleomorphic component. This unusual case ofmetastasizing hepatocellular carcinoma with three distinct histologic components suggests that the histological and immunophenotype of the tumor might be useful in predicting metastatic potential.

Topics: Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Hepatocellular; Diagnosis, Differential; Fatal Outcome; Humans; Immunohistochemistry; Keratins; Liver Neoplasms; Lung Neoplasms; Male; Neoplastic Cells, Circulating

We describe 10 cases of primary well-differentiated neuroendocrine carcinomas (carcinoid tumor) of the lung with extensive sclerotic changes. The patients were 6 women and 4 men from 20 to 69 years of age. Clinically, patients had symptoms of bronchial obstruction such as cough, dyspnea, and chest pain. Surgical resection of the tumors was accomplished in all the cases. Histologically, all tumors corresponded to the well-differentiated type; however, in 4 cases, lymph node metastases were present. Immunohistochemically, all tumors showed positive staining for neuroendocrine markers, including chromogranin, synaptophysin, CD56, and broad-spectrum keratin. Follow-up information showed that 8 patients were alive after a period ranging from 1 to 5 years. The cases presented highlight an important feature of neuroendocrine carcinomas of the lung not previously addressed, one that may pose a problem not only in the diagnosis but also in the grading of these neoplasms.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Neuroendocrine; CD56 Antigen; Chromogranin A; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Sclerosis; Synaptophysin; Treatment Outcome

The purpose of this study was to clarify the role and clinical significance of lymphangiogenesis/micrometastases and adhesion molecules in resected stage I non-small cell lung cancer (NSCLC).. Immunohistochemical (IHC) staining was used to analyze the protein expression of vascular endothelial growth factor-C (VEGF-C), VEGF, E-cadherin, alpha-catenin, beta-catenin, and gamma-catenin in paraffin-embedded tumor samples from 117 well-characterized stage I NSCLC patients and to compare the protein expression, clinical variables and survival outcome. As a micrometastatic parameter in lymph nodes (LNs), cytokeratin (CK) staining was performed.. The positive expression of VEGF-C and VEGF were detected in 54 (48.7%) and 86 (73.5%), respectively. We identified micrometastatic tumor cells in pathological N0 LNs in 34 (29.1%) of 117 patients. E-cadherin, alpha-catenin, beta-catenin, and gamma-catenin were identified in 70 (59.8%), 41 (35.0%), 83 (70.9%), and 61 (52.1%) specimens, respectively. The VEGF-C expression was found more frequently in squamous cell carcinoma (SQ) and in the tumors with negative expression of beta-catenin than counter features. The VEGF expression was found more frequently in the tumors with a negative expression of E-cadherin. Micrometastasis was found more frequently in a pathological T2 status and in the tumors with a negative expression of alpha-catenin. Beta-catenin and gamma-catenin expressions were found less and more frequently in SQ, respectively. A univariate and multivariate survival analysis demonstrated that old age, pathological T2 status, and micrometastasis were independently associated with an increased risk of poor survival in the patients who underwent a surgical resection of stage I NSCLC.. Complicated relationships exist between lymphangiogenesis/micrometastases and adhesion molecules with a specific histology. The detection of lymph nodal micrometastasis by CK may therefore be a useful marker for predicting a poor prognosis in patients who undergo a complete resection of stage I NSCLC.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Cadherins; Carcinoma, Non-Small-Cell Lung; Catenins; Cell Adhesion; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphangiogenesis; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Vascular Endothelial Growth Factors

The aim of the present study was to clarify protein profiling in small cell lung carcinoma (SCLC) and pulmonary large cell neuroendocrine carcinoma (LCNEC). The proteomic approach was used, and involved cell lysate from two cell lines (N231 derived from SCLC and LCN1 derived from LCNEC), with 2-D gel electrophoresis (2-DE). In the present study, 25 protein spots with greater than twofold quantitative differences between LCN1 and N231 cells on 2-DE gels were confirmed. Within the 25 identified proteins, cytokeratins (CK) 7, 8, 18 and 19 were upregulated in LCN1 cells compared with N231 cells. The expression of CK7, 8, 18, and 19 was further studied on immunohistochemistry with 81 formalin-fixed and paraffin-embedded pulmonary carcinomas, which included 27 SCLC, 30 LCNEC, 14 adenocarcinomas, and 10 squamous cell carcinomas. Although the expression of CK7, 8, 18, and 19 was observed in all histological types, the mean immunostaining scores of CK7, 8, 18, and 19 were significantly higher in LCNEC than in SCLC (P < 0.001, P < 0.001, P < 0.01 and P < 0.001, respectively). These data suggest that the biological characteristics of LCNEC and SCLC may be different and the expression of CK may serve as differential diagnostic markers.

Topics: Biomarkers, Tumor; Carcinoma, Large Cell; Carcinoma, Neuroendocrine; Electrophoresis, Gel, Two-Dimensional; Humans; Immunohistochemistry; Keratin-18; Keratin-19; Keratin-7; Keratin-8; Keratins; Lung Neoplasms; Small Cell Lung Carcinoma; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tissue Array Analysis

It can be difficult to distinguish between primary and metastatic squamous cell carcinoma (SCC) in the lung. Surgical specimens were obtained from two groups of patients, 26 lung SCC patients without histories of any other cancer (the definite primary group) and 17 patients who had undergone surgical removal of SCC emerging in the lung after surgery for tongue SCC (the unknown group). From the former, 26 primary lung SCC were obtained. From the latter, 17 lung tumors and 15 primary tumors of the tongue were obtained. Eleven of the 17 lung tumors from the unknown group were metastatic lung SCC. All specimens were immunostained with cytokeratin (CK)5/6, CK7, CAM5.2, CK19 and p63 antibodies. The frequency of CAM5.2 and CK19 expression was significantly higher in the lung SCC of the definite primary group (21 of 26, 81% and 20 of 26, 78%, respectively) than in the metastatic lung SCC (1 of 11, 9% (P < 0.001) and 2 of 11, 18% (P = 0.003), respectively) or primary SCC of the tongue (5 of 15, 33% (P = 0.002) and 2 of 15, 13% (P < 0.001), respectively). CAM5.2 and CK19 are useful for distinguishing between primary SCC of the lung and metastases from tongue cancer.

Topics: Adult; Age Factors; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Chi-Square Distribution; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Tongue Neoplasms

Adjuvant chemotherapy is required following the resection of aggressive NSCLC. It is therefore necessary to identify factors that accurately predict prognosis.. Tumor specimens were collected from 107 patients who underwent a complete resection for NSCLC from 1994-2000 in this Department. The expression of E-cadherin, dysadherin, and cytokeratin in stage I NSCLC specimens was analyzed by immunohistochemistry.. Seventeen percent of tumors showed reduced E-cadherin immunostaining. Twenty-nine per cent of tumors showed dysadherin expression in over 50% of the cancer cells. Positive expression of cytokeratin was identified in 30 (28.0%) patients. The incidence of positive expression of dysadherin in females and elderly patients was higher than that in other patients. Cytokeratin immunoreactive tumor cells in lymph nodes were identified in 34 (28.0%) out of 107 patients. The incidence of positive expression of cytokeratin in T1 tumors was higher than that in T2 tumors. There was a significant inverse correlation between the expression of E-cadherin and dysadherin. The increased expression of cytokeratin was significantly associated with recurrence. Logistic regression models indicated that cytokeratin expression was an independent predictor of recurrence. The increased expression of dysadherin and cytokeratin had a significant impact on patient survival. Furthermore, tumors with an increased expression of dysadherin and a reduced expression of E-cadherin showed the worst prognosis.. The detection of dysadherin in tumors and cytokeratin in the lymph nodes may be a potential significant indicator of a poor prognosis for patients who undergo complete resection of stage I NSCLC.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Cadherins; Carcinoma, Non-Small-Cell Lung; Disease-Free Survival; Female; Humans; Immunohistochemistry; Ion Channels; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Membrane Glycoproteins; Microfilament Proteins; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Prognosis

Phosphoglucose isomerase (PGI) is a multifunctional enzyme that functions in glucose metabolism as a glycolytic enzyme catalyzing an interconversion between glucose and fructose inside the cell, while it acts as cytokine outside the cell, with properties that include autocrine motility factor (AMF)-regulating tumor cell motility. Overexpression of AMF/PGI induces epithelial-to-mesenchymal transition with enhanced malignancy. Recent studies have revealed that silencing of AMF/PGI resulted in mesenchymal-to-epithelial transition (MET) of human lung fibrosarcoma cells and breast cancer cells with reduced malignancy. Here, we constructed a hammerhead ribozyme specific against GUC triplet at the position G390 in the human, mouse, and rat AMF/PGI mRNA sequence. Mesenchymal human osteosarcoma MG-63, HS-Os-1, and murine LM8 cells were stably transfected with the ribozyme specific for AMF/PGI. The stable transfectant cells showed effective downregulation of AMF/PGI expression and subsequent abrogation of AMF/PGI secretion, which resulted in morphologic change with reduced growth, motility, and invasion. Silencing of AMF/PGI induced MET, in which upregulation of E-cadherin and cytokeratins, as well as downregulation of vimentin, were noted. The MET guided by AMF/PGI gene silencing induced osteosarcoma MG-63 to terminally differentiate into mature osteoblasts. Furthermore, MET completely suppressed the tumor growth and pulmonary metastasis of LM8 cells in nude mice. Thus, acquisition of malignancy might be completed in part by upregulation of AMF/PGI, and waiver of malignancy might also be controlled by downregulation of AMF/PGI.

Topics: Animals; Base Sequence; Blotting, Western; Cadherins; Cell Differentiation; Cell Line, Tumor; Cell Movement; Cell Proliferation; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Glucose-6-Phosphate Isomerase; Humans; Keratins; Lung Neoplasms; Molecular Sequence Data; Osteosarcoma; Rats; Reverse Transcriptase Polymerase Chain Reaction; RNA, Catalytic; Sequence Homology, Nucleic Acid; Transfection; Tumor Burden; Vimentin

DeltaNp63 is an isoform of the p53 homolog p63, which lacks an amino-terminal transactivation domain. The aim of this study was to detect the deltaNp63 expression in the squamous carcinoma component of adenosquamous carcinoma and evaluate its usefulness as a specific squamous carcinoma marker.. Immunohistochemistry was used to analyze the protein expression of deltaNp63 and high molecular weight cytokeratin in paraffin-embedded tumor samples from 17 patients with well-characterized adenosquamous carcinoma.. Of 17 cases, 13 (76.5%) and 14 (82.4%) cases showed positive staining for deltaNp63 and HMWCK in the tumor cells, respectively. It was easy to discriminate the squamous carcinoma and adenocarcinoma components in all tumors. Interestingly, positive expression of deltaNp63 was detected in one case with a negative expression of HMWCK.. These findings indicated that the deltaNp63 status was useful for distinguishing squamous carcinoma from adenocarcinoma in formalin-postfixed adenosquamous carcinoma specimens.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Adenosquamous; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Retrospective Studies; Survival Rate; Trans-Activators; Transcription Factors; Treatment Outcome; Tumor Suppressor Proteins

We previously reported a new method of segmentectomy, pulmonary artery-guided segmentectomy as a surgical alternative for small-sized early lung cancer with favorable results, but the follow-up time was too short for definitive conclusion. To examine the efficacy of the segmentectomy, and to determine the appropriate surgical procedure for early lung cancer, we conducted a retrospective follow-up study, and examined the influences of tumor size and preoperative serum tumor marker levels on the prognosis. We reviewed the records of 91 patients who underwent the segmentectomy for pathological T1N0M0 non-small cell lung cancer from 1993 to 2002. In 85 patients, carcinoembryonic antigen, squamous cell carcinoma-related antigen, and a fragment of cytokeratin were measured preoperatively. The overall 5-year survival rate was 83%. Indication (intentional, n=47; compromised, n=44) and tumor size (20mm or less, n=68; 21 to 30 mm, n=23) had no significant impact on survival. The 5-year survival rate for 49 patients with normal tumor marker levels was 93%, and significantly higher than 36 patients with at least one elevated tumor marker level (68%, p<0.01). Median follow-up time of 72.0 months revealed 11 locoregional recurrences. The incidence of locoregional recurrence was significantly higher in the patients with tumors of 21-30 mm, and elevated tumor marker (p<0.01). The follow-up study demonstrated that the segmentectomy could be an acceptable surgical treatment for early lung cancer patients with tumors of 20mm or smaller and normal tumor marker levels.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Disease-Free Survival; Early Detection of Cancer; Female; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Male; Medical Oncology; Middle Aged; Surgical Procedures, Operative; Treatment Outcome

The increasing panel of systemic therapies enables the individual management of cancer patients, even in advanced stages. However, diagnostic tools indicating early the efficacy of therapy are still needed. In prospectively collected sera of 161 patients with recurrent non-small cell lung cancer (NSCLC) receiving second-line chemotherapy, the courses of nucleosomes, cytokeratin-19 fragments (CYFRA 21-1), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and progastrin-releasing peptide (ProGRP) were investigated and correlated with therapy response. At high specificity for detection of progressive disease, most sensitive biomarkers were identified and included in a combination model. High levels and insufficient decreases of nucleosomes and CYFRA 21-1 during the first cycle of therapy indicated poor outcome. Combination of nucleosome concentrations at day 8 and CYFRA 21-1 before start of the second cycle enabled the early detection of progressive disease with a sensitivity of 34.4% at 95% specificity (AUC 0.79) prior to imaging techniques. When cutoffs were fixed at the 90th percentile of responding patients, the combination model achieved sensitivities of 19% at 100% specificity and of 52% at 88% specificity. Thus, nucleosomes and CYFRA 21-1 showed to be valuable for the individual management of patients with recurrent NSCLC.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antineoplastic Agents; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Nucleosomes; Peptides; Phosphopyruvate Hydratase; Protein Precursors; Recurrence

Mucoepidermoid carcinoma (MEC) of the lung needs to be carefully distinguished from other lung tumors with similar features, particularly from adenosquamous carcinoma, this latter tumor frequently showing EGFR mutations. In contrast with the data reported by Han et al in the last July issue of Lung Cancer, neither EGFR nor K-RAS mutations were observed in MEC from caucasian patients.

Topics: Aged; Carcinoma, Mucoepidermoid; Diagnosis, Differential; Epithelium; ErbB Receptors; Female; Genes, ras; Humans; Keratins; Lung Neoplasms; Mucin 5AC; Mucins; Mutation

The current FDA-approved standard of care for nonsmall cell lung cancer is Carboplastin/Taxol/Avastin based upon an impressive survival benefit; however, patients with squamous carcinoma (SQCC) cannot receive Avastin because of a 30% mortality rate due to fatal hemoptysis. In this study we evaluated the role of cytomorphology and immunohistochemistry in differentiating SQCC from adenocarcinoma (ADC) in lung FNA specimens. The case cohort included 53 FNA cases of nonsmall cell lung carcinoma with surgical pathology follow-up. All FNA specimens were reviewed independently by a panel of cytopathologists to differentiate between SQCC and ADC. The cell block material was available in 23 cases (11 ADC and 12 SQCC) to perform immunohistochemical stains for TTF-1, CK7, CK20, P63, and CK5/6. On surgical resection, 35/53 (66%) cases were diagnosed as ADC and 18/53 (34%) as SQCC. The number of cases classified correctly on the basis of cytomorphology was 66% for ADC and 53% for SQCC (combined accuracy 60%). By immunohistochemical staining, 14/23 (61%) cases expressed TTF-1. Nine cases were TTF-1 negative; eight of the TTF-1 negative cases (89%) were SQCC. Twenty-three cases expressed CK7 (87%); one ADC case (4%) showed focal CK20 positivity. Both P63 and CK5/6 expression was seen in 9/12 (75%) SQCC cases; none of the ADC cases showed this dual expression. Cytomorphology alone may not be able to stratify all cases of nonsmall cell lung carcinoma into ADC and SQCC in FNA specimens. The immune-panel of TTF-1, CK7, CK20, P63, and CK5/6 is useful in differentiating SQCC from ADC.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy, Fine-Needle; Carcinoma, Squamous Cell; Diagnosis, Differential; DNA-Binding Proteins; Female; Humans; Keratin-5; Keratin-6; Keratins; Lung Neoplasms; Male; Membrane Proteins; Middle Aged; Transcription Factors

The differential diagnosis of pleural sarcomatoid mesothelioma (SM) from lung sarcomatoid carcinoma (LSC) invading parietal pleura and chest wall is a challenging issue. The aim of this study was to identify useful antibodies that can be used for the differential diagnosis of pleural SM from LSC.. Forty-five cases of pleural SM and 27 cases of LSC were immunohistochemically analysed by using 15 commercially available antibodies, including D2-40 and antibodies to calretinin, thrombomodulin, Wilms' Tumour 1, carcinoembryonic antigen (CEA), Napsin A, thyroid transcription factor (TTF)-1, pan-cytokeratin, CAM5.2, epithelial membrane antigen, Ber-EP4, MOC-31, alpha-smooth muscle actin, h-caldesmon and desmin. The results revealed that D2-40 positivity was significantly higher in pleural SM (86.7%) than in LSC (25.9%). The positivity of the adenocarcinoma markers, including CEA, Napsin A, and TTF-1, was low even in LSC.. Evaluating the positivity and degree of staining of the well-known mesothelial marker D2-40 could be applied to differentiate pleural SM from the sarcomatoid component of LSC, in addition to assessing clinical and radiological information.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Biomarkers; Biomarkers, Tumor; Carcinoma; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mesothelioma; Middle Aged; Pleural Neoplasms

The CXC chemokine, CXCL12, and its receptor, CXCR4 promote metastases of a variety of solid tumors, including non-small cell lung cancer (NSCLC). The expression of CXCR4 on tumor cells may represent a critical biomarker for their propensity to metastasize. This study was performed to evaluate the hypothesis that co-expression of pan-cytokeratin and CXCR4 may be a prognostic marker for patients with advanced NSCLC.. We evaluated CXCR4 levels on circulating pan-cytokeratin positive cells from patients with NSCLC. NSCLC tumor and metastases were also assessed for the presence of CXCR4.. Pan-cytokeratin positive cells were increased in the circulation of patients with NSCLC, as compared to normal control subjects. Patients with pan-cytokeratin +/CXCR4+ = 2,500 cells/ml had a significant improvement in median survival when compared with patients with pan-cytokeratin +/CXCR4+ >2,500 cells/ml (not achieved versus 14 weeks). CXCR4 expression was found on NSCLC tumors and at sites of tumor metastasis.. This study suggests that CXCR4 may be a prognostic marker in NSCLC, and provides hypothesis-generating results, which may be important in determining metastatic potential. In future studies, we will prospectively evaluate the prognostic significance of pan-cytokeratin/CXCR4+ cells, and determine the mechanisms involved in the regulation of CXCR4 expression on tumor cells in a larger patient population.

Topics: Aged; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratins; Lung Neoplasms; Male; Neoplasm Metastasis; Neoplasm Staging; Neoplastic Cells, Circulating; Receptors, CXCR4

Topics: Aged; Antibodies, Monoclonal; Carcinoma, Transitional Cell; Cystectomy; Follow-Up Studies; Humans; Keratin-7; Keratins; Lung Neoplasms; Male; Urinary Bladder Neoplasms; Vimentin

Many evidences suggest that prognosis of non-small cell lung cancer (NSCLC) with lymph node micrometastases (LNMM) is poor compared with those without LNMM. Therefore, it is better to evaluate LNMM through immunohistochemistry (IHC) of serial sectioning of all dissected lymph nodes. However, this labor-intensive approach is impossible in a practical setting. Therefore, we examined whether we are able to efficiently diagnose LNMM using the sentinel node (SN) mapping. Fifty-one patients with clinical T1N0M0 NSCLC were enrolled in this study. SNs were then detected intraoperatively. After SN mapping, lobectomy and hilar and mediastinal lymph node dissection were performed. Metastases of all dissected lymph nodes were examined by hematoxylin and eosin (H&E) staining and immunohistochemical cytokeratin staining. SN detection rate was 80.4% (41/51). Average number of SNs was 1.8+/-1.1 in a patient. Lymph node metastases were diagnosed in two patients using H&E staining. LNMM were found only in SNs of two patients. On the other hand, micrometastasis was not found in non-SN. According to these results, two patients with clinical T1N0M0 NSCLC migrated to T1N1M0. Evaluation of micrometastases of all dissected lymph nodes may be substituted by evaluating micrometastases of SNs. We believe that further studies are warranted to determine the most useful clinical applications.

Topics: Aged; Carcinoma, Non-Small-Cell Lung; Coloring Agents; Eosine Yellowish-(YS); Female; Hematoxylin; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Pneumonectomy; Predictive Value of Tests; Prognosis; Sentinel Lymph Node Biopsy; Staining and Labeling

Targeted therapies have become a valuable therapeutic option for cancer. Establishment of different animal tumor models has become necessary. This study was to establish xenotransplantation models for patient-derived non-small cell lung cancer (NSCLC) in immune deficient mice.. Immune deficient mice, BALB/C-nu, NOD/scid and SCID, 16 in each strain, were used. Sixteen tumor specimens were obtained from patients with NSCLC. Each specimen was subcutaneously transplanted into one mouse from each of the three strains. The tumor formation rate, time to tumor engraftment, tumor volume doubling time were recorded and compared among the three strains of mice. Histology of xenograft tumors was examined.. The total tumor formation rate was 75% (12/16). The tumor formation rate was the highest in SCID mice (56.25%). Only four tumors were engrafted in SCID mice, and two in BALB/C-nu mice. The tumor formation rate, time to tumor engraftment, and tumor volume doubling time were not significantly different among the three strains of mice. The incidence of tumor size over 1cm in the upper flanks of the mice (56.25%) was significantly higher than that in the lower flanks (25%) (P=0.037). Haematoxylin Eosin staining revealed a high degree of histological similarity between all xenograft and the parental tumors.. We have established xenotransplantation models for patient-derived NSCLC with a success rate of 75% in BALB/C-nu and SCID mice. The xenograft tumors have the same histological features to those of their parental tumors.

Topics: Animals; Carcinoma, Non-Small-Cell Lung; Disease Models, Animal; Female; Humans; Keratins; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Inbred NOD; Mice, Nude; Mice, SCID; Neoplasm Transplantation; Transplantation, Heterologous

The report presents a very rare case of papillary adenoma of the lung in a 61-year old man, described for the first time in the Polish literature.

Topics: Adenoma; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Pulmonary Surfactant-Associated Protein A; Respiratory Mucosa

Topics: Adenocarcinoma; Adult; Combined Modality Therapy; Diagnosis, Differential; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Prostate-Specific Antigen; Prostatectomy; Prostatic Neoplasms; Sarcoma, Synovial; Vimentin

Co-expression of cytokeratin and vimentin has been traditionally associated with a few select tumors. However, this phenomenon is being recognized in a wider range of tumors. Twenty-one canine primary pulmonary epithelial neoplasms were evaluated for the co-expression of cytokeratin and vimentin. The histologic pattern and grade, and an immunohistochemical grade for cytokeratin and vimentin staining, were determined for each neoplasm. Adenocarcinomas predominated, and histologically, most tumors were grade II. All of the neoplasms stained positive for cytokeratin, while only 8 (38%) stained positive for both vimentin and cytokeratin. Papillary adenocarcinomas were consistently vimentin negative. The anaplastic histologic pattern had significantly more vimentin staining than the other histologic patterns. There was no significant difference in histologic grade or grading criteria between those tumors that stained with vimentin and those that did not. The present study established that cytokeratin and vimentin co-expression occurs in canine primary pulmonary epithelial tumors at a similar frequency to human pulmonary neoplasms. Further investigation will be needed to characterize the significance of this finding, particularly with respect to prognosis.

Topics: Adenocarcinoma; Animals; Dog Diseases; Dogs; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Necrosis; Vimentin

Tumor-to-tumor metastasis is a very rare event. We report three cases of tumor metastasizing in another tumor: a clear cell renal cell carcinoma in a vesicular thyroid adenoma, a lung carcinoma in a meningioma and a neuroendocrine lung carcinoma in a clear cell renal cell carcinoma. According to the literature, clear cell renal cell carcinoma is the most common tumor recipient of metastasis while lung carcinoma is the most common donor tumor. Several physiopathological mechanisms can explain this phenomenon, but many of them are still unknown.

Topics: Adenocarcinoma, Clear Cell; Aged; Biopsy; Carcinoma, Large Cell; Cell Division; Female; Hemoptysis; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Lung Neoplasms; Male; Meningeal Neoplasms; Meningioma; Middle Aged; Neoplasm Metastasis; Nephrectomy; Thyroid Neoplasms

The purposes of this study were to examine the usefulness of the biopsy of the sentinel lymph nodes (SNs) for the accurate and effective detection of lymph node micrometastasis in early lung cancer and to clarify the spread of lymph node micrometastasis. One hundred and thirty-three c-stage IA non-small cell lung cancer patients in whom SNs could be identified by radioisotope (RI) method were enrolled. All dissected lymph nodes were stained with cytokeratin AE1/AE3 for the examination of micrometastasis. A total of 1375 lymph nodes including 220 SNs were dissected from the 133 patients. From the 220 SNs, 35 (15.9%) were found to be positive for metastasis. Of the other 185 SNs negative for metastasis, 19 (8.6%) were positive for micrometastasis. When patients were limited to those with pN0, there were no lymph nodes positive for micrometastasis other than SNs. In pN1-2 patients, micrometastasis to non-SNs were observed in 2.3-13.2%. In patients with pN0, micrometastasis was limited to SNs, and the results of the examination of SNs for micrometastasis accurately represented those of the examination of all lymph nodes. With advancement of the stage, micrometastasis was not limited to SNs and showed an irregular distribution.

Topics: Carcinoma, Non-Small-Cell Lung; Disease Progression; Early Diagnosis; Female; Humans; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Sentinel Lymph Node Biopsy

Novel targeted treatment of non-small cell lung cancer (NSCLC) requires accurate classification of NSCLC as squamous cell carcinoma (SCC) and adenocarcinoma (AC). This study details the CK5/6 and TTF-1 immunoprofile of surgical resections of 45 NSCLCs (24 ACs and 21 SCCs) in tissue microarrays. All SCCs were CK5/6 positive, TTF-1 negative. 20 of 24 adenocarcinomas had the reverse pattern. In conclusion, all SCCs in this study were CK5/6 positive and TTF-1 negative, and therefore tumours that do not display this phenotype are unlikely to be SCCs. CK5/6 and TTF-1 is therefore a practical panel for the distinction between pulmonary SCC from AC in routine histopathology practice.

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; DNA-Binding Proteins; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Tissue Array Analysis; Transcription Factors

There is no ideal tumour marker at present. The clinical application of CYFRA 21-1 is possible once a thorough standardisation process is carried out. Standardisation is achieved by determining the reference range in asymptomatic population, benign and malignant lung diseases, and benign and malignant diseases of other organs. Furthermore, it depends on knowledge of research population characteristics, patient medical histories and individual diagnostic procedure results, the size of research target samples and the clinically defined control groups. The cut-off level of CYFRA 21-1 for non-small cell lung cancer (NSCLC) is 1.72 ng/mL in the Croatian population. It is based on the clinically applicable sensitivity of 78% and specificity of 95% in benign lung diseases. The cut-off value is verified by clinical findings. For clinicians the level of CYFRA 21-1 is an early sign of NSCLC in relation to all the benign lung diseases and all the benign diseases of other organs, of which it was confirmed that they can influence the above level, provided that NSCLC is verified using standard diagnostic methods. The level of CYFRA 21-1 is also influenced by the time of sampling in relation to other diagnostic invasive procedures. The marker is clinically applicable if clinical findings verify it; otherwise, it is useless. This research has involved 343 healthy persons, 474 patients with a benign disease and 4440 patients with a malignant disease, 2453 of whom suffer from NSCLC. The sensitivity of CYFRA 21-1 in NSCLC is 78%, in squamous cell lung cancer (SQC) 84.6%, in adenocarcinomas (AD) 74.3% and in large cell lung cancer (LCC) 75.3%. The level of CYFRA 21-1 differs significantly between healthy persons, benign and malignant diseases (p<10(-3)). There are differences between the three histological types of NSCLC (p<10(-6)) and according to T and N (p<10(-3)). The level of CYFRA 21-1 prompts clinicians to repeat the clinical procedure during diagnosis, and helps to detect the disease earlier and implement treatment in NSCLC. We have achieved high concordance between marker findings and clinical diagnostic.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Sensitivity and Specificity

To validate the prognostic value of preoperative levels of CYFRA 21-1, CEA and the corresponding tumor marker index (TMI) in patients with stage I non-small cell lung cancer (NSCLC).. Two hundred forty stage I NSCLC patients (80 in pT1 and 160 in pT2; 100 squamous cell carcinomas, 91 adenocarcinomas, 32 large-cell carcinomas, 17 with other histologies; 171 males and 69 females) who had complete resection (R0) between 1986 and 2004 were included in the analysis. CYFRA 21-1 and CEA were measured using the Elecsys system (Roche) and AxSym-System (Abbott), respectively. Univariate analysis was performed using the Kaplan-Meier method to identify potential associations between survival and age, gender, CYFRA 21-1, CEA and TMI.. Overall 3- and 5-year survival rates were 74 and 64%, respectively. Male gender (p = 0.0009) and age >70 years (p = 0.0041) were associated with a worse prognosis; there were no differences between pT1 and pT2 nor between histological subtypes. Three-year survival was 72% for CYFRA 21-1 levels >3.3 ng/ml versus 75% for levels 6.7 ng/ml versus 75% for CEA 0.05). Corresponding 5-year survival rates were near 64% both for patients with CYFRA 21-1 values above and below the cutoff (3.3 ng/ml), and 49 and 66% for patients with values above and below the CEA cutoff (6.7 ng/ml), respectively (both p values >0.05). Overall survival did not vary in the different TMI risk groups (p = 0.73).. In this cohort of early-stage NSCLC patients, male gender and age >70 years were associated with a worse outcome, but elevated levels of CEA and CYFRA 21-1, and TMI risk were not.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Prognosis; Sex Factors; Survival Rate

High circulating serum vascular endothelial growth factor (VEGF) levels might reflect enhanced angiogenesis in patients suffering from non-small cell lung cancer (NSCLC). This study aimed at determining the prognostic significance of circulating VEGF as a prognostic factor in NSCLC.. Four hundred fifty-one histologically or cytologically proven and previously untreated NSCLC patients have been studied. Median follow-up was 13 years and 9 months. Eleven clinical and biologic variables were recorded. The levels of circulating VEGF were measured in the serum by quantitative immunoassay. Patients have had received conventional treatment (without anti-VEGF therapy) according to the international guidelines. All statistical tests were two-sided.. Receiver operating characteristic curves (area under the ROC curve: 0.66 +/- 0.05) showed that circulating VEGF serum level did not demonstrate a high sensitivity-specificity relationship, and therefore, demonstrated a low ability to differentiate NSCLC from benign lung diseases. A 600 pg/mL level of circulating VEGF serum was considered as threshold with 40.8% of NSCLC patients presenting with a high level. The circulating VEGF distribution differed significantly according to disease stage, nodal status, and performance status (PS), with the highest levels observed in metastatic stage, positive mediastinal nodal status, and poor PS. In univariate survival analysis, patients with a high pretreatment circulating VEGF serum level proved to have a shorter overall survival when compared with patients presenting with a circulating VEGF serum level /=2, nodal status N2-3, metastatic disease, neuron-specific enolase >12.5 ng/mL, CYFRA 21-1 >3.6 ng/mL.. The prognostic information given by a high circulating VEGF serum level is not an independent determinant of survival owing to a high relationship with main prognostic variables such as PS, stage of the disease, and nodal status. This finding does not preclude a putative prognostic impact of in situ detection of VEGF and VEGF receptors in tumor specimen.

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Combined Modality Therapy; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Prognosis; Prospective Studies; ROC Curve; Survival Rate; Vascular Endothelial Growth Factor A

To investigate the clinical application value of serum tumor markers detection combined with support vector machine (SVM) model in the diagnosis of oral squamous cell carcinoma.. Serum levels of neuron-specific enolase (NSE), cancer antigen 242 (CA242), cancer antigen 19-9 (CA199), carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA), cancer antigen 72-4 (CA724), cancer antigen 21-1 (CA211) and alpha fetoprotein (AFP) were detected with enzyme-linked immunosorbent assay (ELISA) and time-resolved fluoroimmunoassay (TRFIA) in 163 oral squamous cell carcinoma patients and 160 healthy persons. All the data was analyzed with SVM; the SVM models for diagnosis of oral squamous cell carcinoma were created, trained and validated by cross validation.. Among the 163 oral squamous cell carcinoma patients, there were 128 males and 35 females with the male-to-female ratio of 3.66:1; the age ranged from 30 to 85 years old with a mean age of 59.3 years old; according to the primary site of tumor, 72 cases in tongue, 34 in gingiva, 22 in buccal mucosa, 15 in palatal mucosa, 13 in floor of mouth, 4 in lip and 3 in retromolar region; according to the TNM-UICC classification, there were 33 patients at stage T1, 72 at T2, 44 at T3, 14 at T4, 119 at N0, 42 at N1, 2 at N2, 159 at M0, 4 at M1, 27 at clinical stage I, 51 at stage II, 52 at III, and 33 at IV; according to the pathological differentiation grade, 109 tumors were well differentiated, 42 were moderately differentiated and 12 were poorly differentiated. Five serum tumor markers of CA211, CA199, TPA, CA724 and NSE were selected optimally to create the optimal SVM model for diagnosis of oral squamous cell carcinoma. The accuracy, specificity, sensitivity and positive predictive value of the optimal SVM model were 88.54%, 93.13%, 84.05% and 92.57%, respectively.. From the results, SVM model combined with 5 optimal serum tumor markers is suggested to be used in the diagnosis of oral squamous cell carcinoma. Supported by Shanghai Leading Academic Discipline Project (Grant No.Y0203).

Topics: Adult; Aged; Aged, 80 and over; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Mouth Neoplasms; Phosphopyruvate Hydratase; Sensitivity and Specificity; Support Vector Machine

To investigate the clinical significance of a combination of several serum tumor markers in the diagnosis of lung cancer.. Serum CEA, CA125, CA199, CYFRA21-1 and NSE were measured with RIA, chromatometrychemoluminescence, ELISA and biochemoluminescence methods respectively in 340 patients with lung cancers at different TNM stages, 120 patients with benign lung diseases, and 45 healthy people. The sensitivities, specificities and accuracies of different combination of those markers for the diagnosis of lung cancers were compared.. CYFRA21-1 had the highest sensitivity and accuracy (60.0% and 70.3%) and CA199 had the highest specificity (99.4%) for detecting lung cancers. CYFRA21-1 had the highest sensitivity (79.6%) for detecting squamous carcinoma. CEA had the highest sensitivity of (75.7%) for detecting adenocarcinoma. NSE had the highest sensitivity (76.2%) for detecting small cell lung cancers (SCLC). The combination of several serum tumor markers had higher sensitivities than the single marker for the diagnosis of lung cancers. With two markers, the combination of CYFRA21-1 and NSE had the highest sensitivity and accuracy (82.9% and 83.4%), while the combination of CA125 and CA199 had the highest specificity (94.5%). With three markers, the combination of CEA, NSE and CYFRA21-1 had the highest sensitivity and accuracy (89.1% and 85.1%), while the combination of CEA, CA125 and CA199 had the highest specificity (86.7%). The combination of CEA, CA125, CA199, CYFRA21-1 and NSE produced the best value, with a sensitivity of 93.8%, a specificity of 71.5%, and an accuracy of 86.5%.. Serum CEA, CA125, CA199, CYFRA21-1 and NSE are helpful markers for the diagnosis of lung cancer. The combination of the markers can improve the sensitivity and accuracy of the diagnosis.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; CA-125 Antigen; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Small Cell Lung Carcinoma

Besides new therapeutic drugs, effective diagnostic tools indicating early the efficacy of therapy are required to improve the individual management of patients with nonoperable cancer diseases.. In prospectively collected sera of 128 patients with newly diagnosed small cell lung cancer receiving first-line chemotherapy, the courses of nucleosomes, progastrin-releasing peptide (ProGRP), neuron-specific enolase (NSE), cytokeratin-19 fragments (CYFRA 21-1), and carcinoembryonic antigen were investigated and correlated with therapy response objectified by computed tomography before start of the third treatment course.. In univariate analyses, high levels and insufficient decreases of nucleosomes, ProGRP, NSE, and CYFRA 21-1 during the first and second cycles of therapy correlated with poor outcome. Insufficient response to therapy was most efficiently indicated by the baseline values of nucleosomes, ProGRP, and CYFRA 21-1 before the second therapy cycle reaching areas under the curve (AUC) of 81.8%, 71.3%, and 74.9% in receiver operating characteristic curves, respectively. Combinations of nucleosomes with ProGRP (AUC 84.1%), CYFRA 21-1 (AUC 82.5%), and NSE (AUC 83.6%) further improved the diagnostic power in the high specificity range and yielded sensitivities of 47.1%, 35.3%, and 35.3% at 95% specificity, respectively. In multivariate analyses, including clinical and biochemical variables, only performance score and nucleosomes before cycle 2 were found to independently indicate therapy response.. Biochemical markers specifically identified patients with insufficient therapy response at the early treatment phase and showed to be valuable for diseases management of small cell lung cancer.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antineoplastic Agents; Area Under Curve; Biomarkers, Tumor; Carcinoembryonic Antigen; Drug Resistance, Neoplasm; Female; Humans; Kaplan-Meier Estimate; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Nucleosomes; Peptide Fragments; Phosphopyruvate Hydratase; Prognosis; Recombinant Proteins; ROC Curve; Sensitivity and Specificity; Small Cell Lung Carcinoma

Currently it is necessary to define in almost each case whether a carcinoma is a small or non-small cell carcinoma, adenocarcinoma, pulmonary or metastatic in origin. Thyroid transcription factor-1 (TTF-1) was positive in more than 80% of primary pulmonary adenocarcinomas and in none from the sites other than the thyroid. Mucinous bronchioloalveolar carcinomas are usually negative. Immunocytochemistry with a panel of cytokeratins (CK) 7 and 20, along with TTF-1, is recommended for identification of the origin of adenocarcinoma in pulmonary cytology.. The aim of the study was to assess the value of TTF-1 reactivity in adenocarcinomas determined by immunocytochemistry in different pulmonary cytologic specimens.. Cytologic specimens of 83 patients with adenocarcinomas were analyzed. Immunocytochemistry was performed with a panel of antibodies: TTF-1, CK7, CK20 in all cases and CK5/6 if necessary. The study included 17 different bronchoscopic samples (aspirates, brushes, transbronchial FNA), 14 transthoracic FNA, 27 pleural effusions and 25 FNA of peripheral lymph nodes. TTF-1 was positive in 26/83 (31.3%) and negative in 47/83 (68.7%) samples. All TTF-1 positive adenocarcinomas were also CK7 positive, thus being conclusive of pulmonary origin. In TTF-1 negative group, pulmonary origin was proven in 10/57 (17.5%) adenocarcinomas, whereas 18/57 (31.6%) adenocarcinomas were metastatic; in 29/57 (50.9%) adenocarcinomas other diagnostic procedures failed to prove their origin. CK20 positivity with CK7 negativity was conclusive of metastatic gastrointestinal adenocarcinoma.. Numerous reports support TTF-1 expression in adenocarcinoma as being highly specific for pulmonary origin, if thyroid is excluded. We were able to identify 36/83 (43.4%) adenocarcinomas as pulmonary adenocarcinomas. Among them, only 31.3% were TTF-1 positive. In our study, about 60% of adenocarcinomas with uncertain origin were in the groups of pleural effusions and lymph nodes. In these groups, cytologic diagnosis of adenocarcinoma often provided evidence of the carcinoma expansion, aggressive behavior and poor differentiation, and served as a guideline for patient management. In the studies of mixed pulmonary adenocarcinomas, TTF-1 expression was lower in poorly differentiated segments as well as in the areas with bronchioloalveolar pattern. One explanation for the high percentage of TTF-1 negative adenocarcinomas in our material is morphological selection of adenocarcinomas of presumably non-pulmonary origin before immunocytochemistry.. TTF-1 in a panel with cytokeratins is specific for differentiation of the origin of adenocarcinomas. TTF-1 negative finding in adenocarcinomas does not exclude pulmonary origin, but only points to other diagnostic procedures for definitive diagnosis.

Topics: Adenocarcinoma; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

Squamous cell carcinoma antigen (SCC) is still a widely used tumor marker for monitoring non-small cell lung cancer (NSCLC), although recent reports discourage its routine use because of low sensitivity. This is a study evaluating the efficacy of SCC and CYFRA21-1 in diagnosing NSCLC. A chart review was performed in a university hospital in Japan, covering a period of 10 years, up to October 2004. During the study period, 142 (35.5%) among 400 NSCLC patients diagnosed, received serum assays of both SCC and CYFRA21-1. Elevated SCC and CYFRA21-1 levels were found in 29.6% and 59.2% of patients, respectively. SCC sensitivity was only 13.0% but CYFRA21-1 sensitivity rose to 73.9% in metastatic patients. The adjunct of SCC increased the CYFRA21-1 sensitivity by 6.3% in the overall population and by only 2.2% for patients with metastases. SCC determination should be considered an inefficient method as a potential diagnosing tool for NSCLC patients, and it provides no additional value when used in combination with CYFRA21-1.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Keratins; Lung Neoplasms; Serpins

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Keratins; Lung Neoplasms; Neoplasm Staging; Prognosis

The aim of the study was to analyze the relation between tumor volume (V(path)), tumor marker index (TMI) and prognosis in 261 completely resected (R0) stages I and II non-small cell lung cancer (NSCLC) patients by univariate and multivariate analyses. V(path) was calculated as an ellipsoid body. TMI represents the geometric mean of normalized CYFRA 21-1 and CEA values. Patients with a V(path)< or =13.7cm(3) had a significantly better 5-year-survival rate than patients with a V(path)>13.7cm(3) (78.1% vs. 47.9%; p<0.001). Patients with a TMI< or=0.54 had a 5-year-survival rate of 79.1% compared to only 47.2% in patients with a TMI>0.54 (p<0.001). Besides age (>70 years), performance status and gender, both V(path) (>13.7 cm(3)) and TMI (>0.54) bore significance in the multivariate Cox model with a hazard ratio (HR) of 1.9 (95% CI: 1.1-3.3, p=0.016) and 2.3 (95% CI: 1.3-4.2, p=0.006), respectively. Based on a combination of V(path) and TMI, a low risk group (17% of the patients) with both parameters in the normal range could be identified. Patients with elevated V(path) or TMI (31%) had an intermediate HR of 3.4 (95% CI: 1.3-9.2). When both factors were elevated (52% of patients) the HR increased to 5.95 (95% CI: 2.4-14.9). The elevation of V(path) and TMI was found in 46.2% of stage I and in 59.1% of stage II. The 5-year-survival rates were found to be 89.1, 62.2 and 43.0%, respectively. In conclusion, elevated levels of TMI and V(path) have a strong negative prognostic impact on survival in operated early stage of NSCLC. These patients might be considered for adjuvant chemotherapy.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Follow-Up Studies; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Prognosis; Proportional Hazards Models; Survival Analysis; Tumor Burden

Prognostic implication of serum cytokeratin 19 fragments (CYFRA 21-1) was explored in 60 advanced NSCLC patients, whereas in 45 patients assessable for serological response a >or=35% CYFRA 21-1 decline after two chemotherapy cycles was strongly associated with non-progression (NP), defined as a sum of objective response (OR)+stable disease (P<0.0001) and survival (P=0.0002). Association of OR with survival was not significant. In multivariate survival analysis, >or=35% marker decline and radiological NP status were found as major determinants of prolonged survival with RR: 0.37 (P=0.01) and 0.63 (P=0.01), respectively. In advanced NSCLC patients, NP reflects therapeutic efficacy better than traditional OR. CYFRA 21-1 >or=35% decline seems to be a reliable surrogate marker of treatment efficacy in terms of survival.

Topics: Adenocarcinoma; Adult; Aged; Antigens, Neoplasm; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Prognosis; Prospective Studies; Survival Rate; Treatment Outcome

Krebs von den Lungen-6 (KL-6) is a high molecular weight glycoprotein classified in the category of human MUC1 mucin. KL-6 has been reported to serve as a sensitive marker for interstitial pneumonia; however, recent studies have suggested that it can also be used as a tumor marker as its origin shows. To further elucidate the clinicopathological significance of circulating KL-6 in lung cancer, we monitored the circulating KL-6 levels in advanced nonsmall cell lung cancer (NSCLC) patients and analyzed the association between these levels and the clinical outcome of EGFR-TKI treatment. The pretreatment levels of circulating KL-6 were found to be significantly higher in progressive disease (PD) patients than disease-controlled (partial response (PR) and stable disease (SD)) patients. Multivariate analyses revealed the circulating KL-6 level to be an independent prognostic factor for overall survival as well as progression-free survival. In addition to these observations, we found that changes in circulating KL-6 levels at 2 weeks after the start of EGFR-TKI treatment from the baseline could quite precisely discriminate PD cases from PR or SD patients and the clinical outcome of EGFR-TKI in NSCLC patients. These results indicate that the monitoring of circulating KL-6 levels in NSCLC patients is effective for both selecting patients to be treated with EGFR-TKI and predicting the clinical outcome of EGFR-TKI. In addition, the findings suggest that the circulating KL-6 level could be used as a clinically relevant biomarker in patients with NSCLC, particularly those who are candidates for EGFR-TKI treatment.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antineoplastic Agents; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Disease Progression; Disease-Free Survival; Electrochemistry; ErbB Receptors; Female; Humans; Keratin-19; Keratins; Luminescent Measurements; Lung Neoplasms; Male; Middle Aged; Mucin-1; Mutation; Odds Ratio; Patient Selection; Predictive Value of Tests; Proportional Hazards Models; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Treatment Outcome

Determination of tumor biomarkers in serum has been proposed as an alternative and noninvasive way of establishing diagnosis for lung cancer. The aim of this study was to assess the combined use of Cyfra 21-1; the soluble fragment of cytokeratin 19, and soluble E-selectin; endothelial adhesion molecule, for improving the accuracy of diagnosing and screening lung cancer.. Serum Cyfra 21-1 and sE-selectin were determined using electrochemiluminescent immunoassay and enzyme linked immunosorbent assay, respectively, in 52 patients with histologically proven lung cancer, 40 patients with benign lung diseases and 50 healthy volunteers.. Both markers were significantly increased in lung cancer patients as compared to healthy, for benign lung diseases Cyfra 21-1 perform well than sE-selectin. Sensitivities of Cyfra 21-1 and sE-selectin at 95th percentiles of the normal group were 96.2% and 92.3%, while at 95th percentiles of the benign lung diseases were 57.7% and 5%, respectively. In ROC curves, the diagnostic power for the detection of lung cancer was similar for Cyfra 21-1 and sE-selectin; however, when compared with benign lung diseases, Cyfra 21-1 was superior to sE-selectin. The highest sensitivity (99.8%) was reported when the two markers were combined.. Cyfra 21-1 and sE-selectin show good performance in detecting lung cancer from normal groups, however, Cyfra 21-1 was superior to sE-selectin in discriminating lung cancer from benign lung diseases.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; E-Selectin; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratin-19; Keratins; Luminescent Measurements; Lung Neoplasms; Male; Middle Aged; Sensitivity and Specificity

The patient was a 63-year-old man who consulted our hospital with complaints of a cough and breathing difficulties. His chest CT revealed a 25-mm mass in his right S1 hilar area with spiculation, disseminated nodule in right lung, and pericardial effusions. Also, bronchoscope and TBLB revealed squamous cell carcinoma. This patient was diagnosed as lung cancer (cT4N3M1, stage IV), and chemotherapy was initiated. The chemotherapy was given in the order of CBDCA (AUC3) +GEM (1,000 mg/m(2)), DOC (60 mg/m(2)), and VNR (25 mg/m(2)), and the tumor response was PD. S- 1 (120 mg/body/day, continuous administration for 2 weeks followed by 1 week of rest) was chosen as fourth-line treatment, and a breast CT detected tumor size reduction following completion of the first course. However, after completion of three courses, the breast CT found tumor-enlargement again. Then the chemotherapy was changed to amrubicin (35 mg/m(2)), but the treatment was discontinued due to skin rash. We once experienced a size reduction with S-1, so S-1 (100 mg/body/day, day 1-14) plus CPT-11 (60 mg/m(2), day 1, 7, 14) combination chemotherapy was conducted at 4-week intervals. After two courses were completed, tumor size reduction was observed by breast XP and CT. The response rate was 40.0%. Currently, seven courses were completed, and we will continue this treatment due to the tumor response of SD. The S-1 single treatment and S-1+CPT-11 combination chemotherapy showed efficacy for this difficult case of NSCLC with refractoriness to multiple cancer drug chemotherapy. This combination treatment should be investigated further for its therapeutic benefit.

Topics: Antigens, Neoplasm; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Camptothecin; Carcinoma, Squamous Cell; Drug Combinations; Drug Resistance, Neoplasm; Humans; Irinotecan; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Oxonic Acid; Tegafur; Tomography, X-Ray Computed

To present an alternative method of detecting micrometastases in lymph nodes previously testing negative for non-small cell lung cancer (NSCLC) by routine hematoxylin-eosin staining.. A total of 77 hilar and mediastinal lymph nodes resected from 18 patients with NSCLC were investigated for the presence of micrometastases using a combination of microarray analysis and immunohistochemistry.. Micrometastases were detected by identifying cytokeratin- and chromogranin-positive cells in lymph node microarrays. Of the 18 patients initially staged as pN0 through routine hematoxylin-eosin staining, 9 (50%) were restaged as N1, and the prognoses were re-evaluated in terms of histological and clinical parameters. The comparison of the survival curves revealed that survival was higher in the patients without micrometastases than in those with micrometastases. In addition, in the multivariate analysis adjusted for age, gender, histological type, and restaging, the presence of micrometastases proved to be an independent predictor of survival. Among patients who had been previously staged as pN0, the risk of death was found to be 7-times greater for those later diagnosed with micrometastases than for those in whom no micrometastases were identified.. The combination of microarray analysis and immunohistochemistry might represent a low-cost and less time-consuming alternative for identifying occult micrometastases and predicting prognoses in surgically resected patients with pN0 NSCLC. Larger randomized, prospective studies are needed in order to determine the accuracy of this method.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Brazil; Carcinoma, Non-Small-Cell Lung; Chromogranin A; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Male; Microarray Analysis; Middle Aged; Neoplasm Staging; Prognosis

The phospholipid fatty acid profiles of erythrocytes and platelets from fifty patients with advanced non-small cell lung cancer were investigated using gas chromatography/mass spectrometry, followed by "ROC" curves analysis to gain novel biomarker information. Sialic acid and cytokeratins were also examined. Potentially useful fatty acid markers: Erythrocytes: phosphatidylcholine, 18:2n6 and 20:4n6; phosphatidylethanolamine, 22:4n6 and 22:6n3 + 24:1n9. Platelets: phosphatidylcholine, 22.0; phosphatidylethanolamine, 22:5n3 + 24:0. At the cut-off value to obtain maximum accuracy, the best biomarkers were found in platelets: phosphatidylserine + phosphatidylinositol (PS + PI), 21:0; sphyngomyelin: 20:1n9 and 22:1n9. All these fatty acids showed similar/higher diagnostic yields than the commonly used markers sialic acid or cytokeratins.

Topics: Adenocarcinoma; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Blood Platelets; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Erythrocytes; Fatty Acids; Female; Gas Chromatography-Mass Spectrometry; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; N-Acetylneuraminic Acid; Neoplasm Proteins; Peptides; Phospholipids; ROC Curve; Sensitivity and Specificity

This study aimed to establish the clinical significance of preoperative serum cytokeratin 19 fragment (CYFRA21-1) and sialyl-Lewis x (SLex) as prognostic markers.. The study involved 272 patients (181 male, 91 female; median age 69 years; range, 32 to 92) with non-small cell lung cancer (NSCLC) who underwent pulmonary resection with mediastinal lymph node dissection. Tumor markers carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), CYFRA21-1, and SLex were examined.. A log-rank test revealed that age, gender, performance status, CEA, SCC, CYFRA21-1, and SLex were associated with the survival rate. By multivariate analysis, age, gender, performance status, CYFRA21-1 (risk ratio, 2.42) and SLex (risk ratio, 6.18) were independent prognostic factors. For patients positive for both markers, the relative risk was 6.10 compared with patients negative for both markers. The patients were divided into three groups: negative for both CYFRA21-1 and SLex (n = 97); positive for either marker (n = 136); and positive for both markers (n = 39). The 1-, 3-, and 5-year survival rates were the following: 98%, 82%, and 75% in the first group; 90%, 63%, and 49% in the second group; and 62%, 31%, and 25% in the third group (p < 0.001). Sixty-four percent of patients positive for both markers were histologic stage III/IV, and 68% of patients negative for both markers were stage I.. Serum CYFRA21-1 and SLex were prognostic markers for NSCLC. Their combination should contribute to the classification of NSCLC patients. Preoperative staging should be carefully performed in patients positive for both tumor markers.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Keratins; Lung Neoplasms; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen

The aim of this study was to evaluate the individual and combined diagnostic value of five tumour markers in the elderly patients with pleural effusions. Serum and pleural fluid levels of cytokeratin fragment 19 (CYFRA21-1), neuron-specific enolase (NSE), carbohydrate antigen 15-3 (CA15-3), carbohydrate antigen 19-9 (CA19-9) and carbohydrate antigen 125 (CA125) were assayed in 32 elderly patients with malignant pleural effusions resulting from advanced lung cancer and in 30 elderly patients with benign pleural effusions by ELISA. Serum levels of CYFRA21-1, NSE, CA15-3, CA19-9 and CA125 in patients with malignant pleural effusions were 12.84 +/- 6.48 microg/l, 22.07 +/- 11.25 microg/l, 65.74 +/- 30.26 kU/l, 56.32 +/- 25.6 kU/l and 71.86 +/- 31.45 kU/l, respectively, and were significantly higher than those in patients with benign pleural effusions (p < 0.01). Pleural fluid levels of CYFRA21-1, CA15-3, CA19-9 and CA125 except NSE in patients with malignant pleural effusions were 18.64 +/- 8.15 microg/l, 59.31 +/- 27.35 kU/l, 48.24 +/- 21.56 kU/l and 62.16 +/- 27.79 kU/l, respectively, and were significantly higher than those in patients with benign pleural effusions (p < 0.01). The parallel combined testing of five tumour markers in serum increased the diagnostic sensitivity to 90.6%, and serial combined testing increased the diagnostic specificity to 93.3%. The sensitivity (%) and specificity (%) of these tumour markers in pleural fluid were as follows: CYFRA21-1, 84.4/90; CA15-3, 62.5/73.3; CA19-9, 37.5/66.7; CA125, 56.3/70; for differentiating malignant effusions from benign effusions. When CYFRA21-1 and CA15-3 combined, the sensitivity and specificity were increased (100% and 90% respectively). Serum and pleural fluid levels of the five tumour markers shows certain values in the diagnosis and differentiate diagnosis for malignant pleural effusions in the elderly patients from benign. The combined assay of five tumour markers in serum and the CYFRA21-1 combined with CA15-3 in pleural fluid were helpful and can increase the sensitivity and specificity in diagnosing malignant pleural effusions.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; CA-125 Antigen; CA-19-9 Antigen; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Mucin-1; Phosphopyruvate Hydratase; Pleural Effusion; Sensitivity and Specificity

Basaloid squamous cell carcinoma of the lung, an uncommon subtype of non-small cell carcinomas was introduced as a distinct entity in the recently revised World Health Organization (WHO) classification of lung tumours. This rare tumour most commonly develops in males older than 60 years. We report a 23-years-old female patient with basaloid squamous cell carcinoma of the lung who was stage IIB post-operatively. The patient is still alive and healthy 18 months after the operation. This is one of the youngest patient reported with this rare type of tumour.

Topics: Adult; Bronchoscopy; Carcinoma, Squamous Cell; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Neoplasm Staging; Radiography; Time Factors; Treatment Outcome

Primary pulmonary and mediastinal synovial sarcoma is rare and poses a diagnostic challenge particularly when unusual histological features are present. We present 60 cases of primary pulmonary and mediastinal synovial sarcoma (29 male and 27 female subjects; mean age, 42 years) and compare our results with five prior series to better define unusual histological features. Clinically, patients with mediastinal synovial sarcoma were younger with a male gender bias. Radiologically, tumors were well delineated with distinctive magnetic resonance imaging features and little vascular enhancement. In all, 21/46 patients died of disease within 5 years. Histologically, all tumors had dense cellularity, interlacing fascicles, hyalinized stroma, and mast cell influx. Hemangiopericytoma-like vasculature (48/60), focal myxoid change (30/60), and entrapped pneumocytes (23/60) were seen. Calcification was not prevalent (10/60). Unusual histological features included Verocay body-like formations (7/60), vague rosettes (6/60), well-formed papillary structures (3/60), adenomatoid change (3/60), and rhabdoid morphology (2/60). Immunohistochemistry demonstrated expression of pancytokeratin (39/58), epithelial membrane antigen (29/53), cytokeratin 7 (26/40), cytokeratin 5/6 (5/7), calretinin (15/23), CD99 (19/23), bcl-2 (24/24), CD56 (11/11), S-100 (9/51), and smooth muscle actin (8/32). In total, 92% (36/39) of primary pulmonary and mediastinal synovial sarcomas studied were positive for t(x;18). In conclusion, our study confirms the clinical, histological, immunohistochemical, and molecular data from previous large series of primary pulmonary and mediastinal synovial sarcoma. Compared with soft tissue synovial sarcoma, primary pulmonary and mediastinal synovial sarcoma has less calcification, less obvious mast cell influx, and less radiologic vascularity, but similar magnetic resonance imaging features, percentage of poorly differentiated tumors, and number of t(x;18)-positive tumors. Awareness of focal unusual histology can prevent misdiagnosis particularly in t(x;18)-negative tumors.

Topics: 12E7 Antigen; Adult; Antigens, CD; Calbindin 2; CD56 Antigen; Cell Adhesion Molecules; Chromosomes, Human, Pair 18; Chromosomes, Human, X; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Mediastinal Neoplasms; Oncogene Proteins, Fusion; Proto-Oncogene Proteins c-bcl-2; S100 Calcium Binding Protein G; S100 Proteins; Sarcoma, Synovial; Survival Rate; Translocation, Genetic

Topics: Adenocarcinoma; Aged; Carcinoma, Papillary; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Keratin-7; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Mucin-1

Although distinguishing metastatic colorectal adenocarcinoma from primary lung adenocarcinoma is often difficult, pre- or intra-operative identification is very important, as the resection areas for each diagnosis differ substantially. CDX2, a recently cloned homeobox gene, represents a highly specific and sensitive marker of colorectal adenocarcinoma. We evaluated CDX2 expression using pre- and intra-operative biopsy specimens. The study examined 50 consecutive colorectal adenocarcinoma metastases to the lung, including 20 biopsy specimens and 66 resected specimens, and 21 primary lung adenocarcinomas. All specimens were immunohistochemically stained for CDX2, cytokeratin (CK) 7, CK20 and thyroid transcription factor (TTF)-1, and scored in a semi-quantitative manner. Mean staining score in biopsy specimens was significantly higher for CDX2 than for CK20. Sensitivities for CDX2 and CK7-/20+ in biopsy specimens were 95.0 and 65.0%, respectively. If CDX2 immunostaining had not been performed, 8 biopsy specimens (40%), and 20 resected specimens (30.3%) might have been diagnosed as equivocal cases either as primary lung cancer or metastatic colorectal cancer, using other markers. These results suggest that positive CDX2 staining represents a highly sensitive and specific marker of metastatic colorectal carcinoma in both biopsy and resected specimens, and is superior to staining for the CK7-/20+ phenotype.

Topics: Adenocarcinoma; Biomarkers, Tumor; Biopsy; CDX2 Transcription Factor; Colorectal Neoplasms; Homeodomain Proteins; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Phenotype; Preoperative Care; Sensitivity and Specificity

A reliable diagnosis of small cell lung cancers (SCLC) is of high clinical relevance. We investigated whether immunocytology substantially improves the diagnostic accuracy of conventional cytology in diagnosing SCLC.. 162 carcinomatous specimens clinically suspected to originate from pulmonary neoplasms were investigated by cytology and immunocytology. Immunocytology was performed on smears using HEA125 and pancytokeratin antibodies as epithelial markers and MOC-1 as SCLC probe.. As histologically clarified, 114 specimens corresponded to pulmonary neoplasms (SCLC = 51; non-small cell lung cancer: NSCLC = 59; mixed SCLC/NSCLC = 2; carcinoid = 2), 48 to nonpulmonary adenocarcinomas. By conventional cytology tumor cells were clearly detected in 93 (57.4%) and suspected in another 43 (26.5%) cases (83.9% overall sensitivity). Considering SCLC samples, tumor cells were diagnosed or suspected in 36 (70.5%), not identified in 10 (19.6%), and misdiagnosed as hematological malignancy in 5 cases. Only 2 specimens were accurately diagnosed as SCLC. Using the epithelial antibodies all samples were identified as carcinomatous. MOC-1 stained all but one SCLC, both SCLC/NSCLC, and both carcinoids. One SCLC brush smear was MOC-1 negative, containing only squamous epithelium. 3 pulmonary adenocarcinomas stained falsely positive, all nonpulmonary carcinomas MOC-1 negative.. Immunocytology substantially improves the diagnostic accuracy of cytology in diagnosing SCLC with a diagnostic sensitivity of 98% and specificity of 97%.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoid Tumor; Carcinoma, Small Cell; Carcinoma, Squamous Cell; CD56 Antigen; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Lung; Lung Neoplasms; Predictive Value of Tests

Reverse transcriptase-polymerase chain reaction assay results have indicated that tumor cells sometimes appear during surgery for primary non-small-cell lung cancer. In this study, we attempted to determine whether cancer cells can be detected during and after surgery using an immunocytology method.. Nine patients undergoing a lobectomy for non-small-cell lung cancer were studied. The presence of circulating tumor cells was determined by the detection of magnified EpCAM antibodies. The criteria used to identify circulating tumor cells were a round-to-oval morphology with a visible nucleus (4'-6'-diamidino-2-phenylindole (DAPI)-positive), which were positive for cytokeratin and negative for CD45.. One patient showed evidence of circulating tumor cells at thoracotomy, and 3 patients did so after surgery. Ten days after the operation, the circulating tumor cells had disappeared in all these cases. The median follow-up period was 14 months, and there was no cancer recurrence in any of the patients.. Using this technique, tumor cells were detected in the peripheral blood of patients before and after lobectomy procedures. It could be argued that this method can provide useful information about patients undergoing lung cancer treatment.

Topics: Aged; Antibodies, Neoplasm; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Case-Control Studies; Cell Adhesion Molecules; Epithelial Cell Adhesion Molecule; Follow-Up Studies; Humans; Immunohistochemistry; Intraoperative Period; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Neoplastic Cells, Circulating; Pilot Projects; Postoperative Period; Reverse Transcriptase Polymerase Chain Reaction; Thoracotomy; Treatment Outcome

The aim of this retrospective study was to assess the prognostic value of serum tumor markers (carcinoembryonic antigen (CEA) and CYFRA21-1) in patients with pathologic (p-) stage I non-small cell lung cancer (NSCLC) undergoing complete resection.. Two hundred and seventy-five patients (163 males, 112 females, mean age 67.1 years) with p-stage I NSCLC who underwent complete resection at our institution between April 1999 and October 2004 were examined. Patients who had received preoperative chemotherapy or radiotherapy were excluded, as were patients who had multiple malignancies including multiple lung cancer. The serum levels of tumor markers were measured using commercially available immunoassays within 1 month before surgical resection. Serum levels of CEA and CYFRA21-1 higher than 5.0 and 2.8 ng/ml, respectively, were considered as positive according to the manufacture's instructions.. The histological classification was adenocarcinoma in 193 patients, squamous cell carcinoma in 71, large cell carcinoma in 5, and other histological type in 6. One hundred and fifty-seven patients had T1 disease and 118 patients had T2 disease. The positive ratio of CEA and CYFRA21-1 was 25.7% and 13.7%, respectively, and in relation to histological type was 27.8% and 7.8% in adenocarcinoma, and 20.6% and 28.4% in squamous cell carcinoma. The overall 5-year survival rate was 79.3%. With a median follow-up of 35.5 month for surviving patients, those with initial CYFRA21-1 serum levels higher than 2.8 ng/ml had a significantly worse prognosis (p=0.0041). Patients with an elevated preoperative CEA level exceeding 5.0 ng/ml had a shorter disease-free survival period (p=0.0003). In patients with adenocarcinoma, a CEA level above 5.0 ng/ml was associated with shorter survival and early recurrence, whereas CYFRA21-1 showed no such association. In patients with squamous cell carcinoma, elevated preoperative CEA was not related to survival and recurrence. In these patients, preoperative CYFRA21-1 level exceeding 2.8 ng/ml was associated with a poorer outcome, whereas preoperative CYFRA21-1 level was not associated with cancer recurrence.. The patients with p-stage I adenocarcinoma whose preoperative CEA level was high might be considered as good candidates for adjuvant chemotherapy. The prognostic value of CYFRA21-1 could not be confirmed for stage I NSCLC, and preoperative CYFRA21-1 level was not useful in selecting the candidates for adjuvant chemotherapy.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Disease-Free Survival; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Prognosis; Pulmonary Surgical Procedures; Regression Analysis; Retrospective Studies; Survival Analysis

The 2004 WHO classification of lung tumours recognised basaloid carcinoma as a variant of squamous and large cell carcinoma. We report a unique case of primary pulmonary adenocarcinoma with a basaloid component. An 82-year-old man underwent pulmonary lobectomy for a 2.8 cm tumour. The patient is disease-free 13 months after diagnosis. Histologically, an invasive carcinoma having a glandular and a solid component was observed. The former was an adenocarcinoma with mucus containing spaces lined by columnar mucinous cells and basaloid cells. The solid component was an organoid proliferation of basaloid-type cells, as in cutaneous basal cell carcinoma. Basaloid cells, but not mucinous cells, were immunoreactive for high molecular weight cytokeratins (CK), CK 7 and, focally, for TTF-1. High Ki67 index, p53 and EGFR expression were also found. This tumour is unique in several respects: (1) The solid areas resemble a conventional basaloid carcinoma, except for the presence of small mucin-containing spaces. (2) The mucinous adenocarcinoma areas contain two layers of columnar and basaloid cells. (3) Both components are neoplastic based on cell morphology, invasive properties and phenotypic profile. These findings indicate that a basaloid variant of adenocarcinoma is also existing in the spectrum of basaloid carcinomas of the lung.

Topics: Adenocarcinoma; Aged, 80 and over; Carcinoma, Basal Cell; Humans; Keratins; Lung Neoplasms; Male; Mucins; Neoplasm Invasiveness; Phenotype; World Health Organization

For deveplopment and function of the lung, progesterone (Prog) fulfils important roles. In a recent report, immunolocalization of Prog and estrogen receptors in non-small cell lung carcinomas were examined and it was shown that the Prog receptor might be a potent prognostic factor. In the present study, a cell line with the sensitivity to Prog was established from a human lung cancer and the growth mechanism was analyzed. The proliferation of established SN96-42 cells was sensitive to Prog and antiprogesterone RU38486 inhibited their proliferation stimulated by Prog. Exposure of these cells to Prog resulted in a decreased formation of leukotriene (LT). The 5-lipoxygenase inhibitor (5-LOX), AA861, effectively stimulated SN96-42 cell proliferation and 5-LOX-catalyzed product(s), especially LTC4, inhibited SN96-42 cell proliferation caused by Prog. Prog-sensitive enhancement of SN96-42 cell proliferation is at least partly mediated through an inhibition of LT formation and these data suggest that 5-LOX and LTs play important roles in SN96-42 cell proliferation stimulated by Prog.

Topics: Carcinoma, Squamous Cell; Cell Division; Cell Line, Tumor; Humans; Keratins; Leukotrienes; Lung Neoplasms; Progesterone; Receptors, Estrogen; Receptors, Progesterone

This study aimed to establish the clinical significance of preoperative serum cytokeratin 19 fragment (CYFRA21-1) and Sialyl Lewis(x) (SLX) in patients with stage I non-small cell lung cancer (NSCLC). The study involved 137 patients (87 male, 50 female; median age 69 years) with completely resected stage I NSCLC. SLX, carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), and CYFRA21-1 were examined. Receiver operator characteristic (ROC) curves were constructed to determine prognostic cut-off values. Among the 137 patients, we identified 30 with recurrence within 3 years. The 5-year survival rates in patients with (n=30) and without (n=107) recurrence were 14% and 81%, respectively. The serum concentrations of SLX, CEA, and CYFRA21-1 in the recurrence group were significantly higher than those in the non-recurrence group. The areas under the ROC curve (AUC) were 0.72, 0.65, 0.53, and 0.64 for SLX, CEA, SCC, and CYFRA21-1, respectively. The prognostic cut-off values were 36U/ml, 7.8ng/ml, 1.5ng/ml, and 3.2ng/ml for SLX, CEA, SCC, and CYFRA21-1, respectively. A log-rank test revealed that age, performance status, T factor, lymphatic invasion, vascular invasion, SLX, CEA, SCC, and CYFRA21-1 were all significantly associated with survival. By multivariate analysis, age, performance status, lymphatic invasion, SLX (risk ratio, 4.11) and CYFRA21-1 (risk ratio, 3.47) were independent prognostic factors. For patients positive for both CYFRA21-1 and SLX, the relative risk was 5.32 compared with patients who were negative for both markers. The 5-year survival rates were 80% in the group negative for both markers (n=86); 52% in the group positive for one of the markers (n=43); and 13% for the group positive for both markers (n=8) (p<0.001). We concluded that serum SLX and CYFRA21-1 were prognostic markers for stage I NSCLC. Their combination should contribute to the classification of stage I NSCLC patients. There is a need to consider adjuvant and neoadjuvant therapies to improve prognosis in patients positive for both tumor markers.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Oligosaccharides; Peptide Fragments; Sialyl Lewis X Antigen; Survival Rate; Time Factors; Treatment Outcome

The clinical importance of preoperative CYFRA 21-1 measurement in early-stage non-small cell lung cancer (NSCLC) is still unclear. The aim of this study is to clarify the prognostic value of preoperative CYFRA 21-1 levels in clinical stage (c-stage) I NSCLC.. The records of 101 c-stage I NSCLC patients who had undergone complete resection were analyzed to correlate preoperative CYFRA 21-1 levels to both the pathologic factors of resected specimens and postoperative outcomes. The cut-off value was set at 3.5 ng/ml.. Six cases (5.9%) showed high CYFRA 21-1 (> or =3.5 ng/ml). The 5-year survival of normal and high CYFRA 21-1 groups was 83.3% and 50.0%, respectively. Patients with high CYFRA 21-1 had significantly poor outcomes (P=0.006). In univariate analysis, preoperative serum CYFRA 21-1 level, pT, pN, and p-stage were significantly associated with prognosis. Multivariate analysis showed that only CYFRA 21-1 level was retained as an independent prognostic factor (relative risk=9.79, P=0.002).. CYFRA 21-1 is an independent predictor of poor outcome for c-stage I NSCLC. Elevated preoperative CYFRA 21-1 levels in early-stage NSCLC may indicate a subgroup at high risk of early death, which has the potential for better survival with additional systemic chemotherapy.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Neoplasm Staging; Prognosis; Survival Analysis; Survival Rate

The purpose of this study is to evaluate the value of cytokeratin (CK) MNF116 and vimentin in the differential diagnosis of malignant pleural effusions. There were evaluated smears from 30 patients with pleural effusions stained with May-Grünwald Giemsa and Papanicolaou techniques for the routine cytological diagnosis. Additional smears were immunostained with CK MNF116 and vimentin using LSAB2 technique. Two independent observers evaluated all smears. Smears were classified first by cytological examination in seven cases (23.33%) as benign, and in 23 cases (76.67%) as malignant pleural effusions. Mesothelial cells expressed CK MNF116 in 96.67% (29/30) of cases and vimentin in 33.33% (10/30) of cases. Malignant cells expressed CK MNF116 in 52.17% (12/23) of cases and vimentin in 30.43% (7/23) of cases. The pattern of immunostaining was diffuse cytoplasmic. In conclusion, CK MNF116 and vimentin may be used as a part of the panel of antibodies for differential diagnosis of malignant pleural effusions with primary unknown.

Topics: Adenocarcinoma; Antibodies; Carcinoma, Small Cell; Diagnosis, Differential; Eosine Yellowish-(YS); Female; Humans; Keratins; Lung Neoplasms; Lymphoma, Large B-Cell, Diffuse; Melanoma; Methylene Blue; Pleural Effusion, Malignant; Pleural Neoplasms; Vimentin

Breast cancer, a whole body disease, can metastasize at early stage. This study was to explore the correlation of peripheral blood cancer cell (PBCC) content to distant metastasis of breast cancer.. The PBCC content of 65 breast cancer patients and 8 healthy donors was detected by multi-parameter flow cytometry (FCM) with CD45 and cytokeratin staining.. Cancer cells were detected in peripheral blood samples from 57 of the 65 patients; the positive rate was 87.7%. No cancer cell was found in peripheral blood samples from healthy donors. The positive rate of PBCCs was correlated to T stage (r=0.271,P=0.017) and N stage (r=0.393, P=0.002). The patients were followed for 5 years; 2 were lost. Distant metastasis was found in 25 patients with PBCCs. In contrast, no metastasis was found in 8 patients without PBCCs (P<0.05).. Preoperative PBCC content is closely related to distant metastasis of breast cancer. The detection of PBCCs might be useful for individual treatment decision for breast cancer.

Topics: Adult; Aged; Breast Neoplasms; Carcinoma; Female; Fibroma; Flow Cytometry; Follow-Up Studies; Humans; Keratins; Leukocyte Common Antigens; Liver Neoplasms; Lung Neoplasms; Middle Aged; Neoplasm Staging; Neoplastic Cells, Circulating; Young Adult

To investigate the clinicopathologic features of tumors showing perivascular epithelioid cell differentiation (PEComas).. The clinicopathologic data of 39 cases pf angiomyolipoma (AML), 17 males and 22 females, with the primary focus in the kidney on 30 cases, in the liver in 4 cases, in the lung, uterus and broad ligament, abdominal wall, retroperitoneum, and nasal cavity in 1 case respectively, were analyzed. Immunohistochemistry (HIC) was used to detect the expression of pan-cytokeratin (CK), S-100 protein, smooth muscle actin (SMA), desmin, vimentin, HMB45, Melan-A, microphthalmia transcription factor (MiTF), CD117, and CD34 in the specimens of the tumors obtained during operation. Twenty patients were followed up.. Pathological examination showed branched capillaries or arterioles, often thick-walled similar to those in the renal cell carcinoma, and the cancerous cells consisting of the mixture of epithelial cells and spindle cells. HIC showed that the expression rates of Melan-A, HMB45, MITF, SMA, desmin, S-100 protein, vimentin, CD117, CK, and CD34 were 95% (37/39), 72% (32/39), 46% (18/39), 82% (32/39), 27% (10/39), 15% (6/39), 82% (32/39), 10% (4/39), 0, and 0 respectively. Clinical follow-up showed 1 patient alive with tumor, and 19 alive free from disease.. PEComas have distinctive morphological and immunohistochemical features.

Topics: Adult; Aged; Angiomyolipoma; Antigens, CD34; Cell Differentiation; Desmin; Epithelioid Cells; Female; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Nose Neoplasms; Proto-Oncogene Proteins c-kit; Retrospective Studies; S100 Proteins; Vimentin

We report a unique case of adenoid cystic carcinoma (ACC) of the maxillary sinus, with gradual histologic transformation from lower-grade ACC (cribriform and tubular types) to high-grade adenocarcinoma (HGA) showing a sequential histologic spectrum via solid-type ACC. A 74-year-old man presented with swelling and mild pain of the right cheek. CT scan showed a mass measuring approximately 4 cm, with marked bone destruction in the right maxillary sinus. A surgically resected specimen revealed that the tumor was comprised of three different components: HGA and solid-type ACC in the central portion and lower-grade ACC in the periphery. The tumor was discriminated from a dedifferentiated carcinoma or hybrid tumor. Autopsy specimens also demonstrated both solid-type ACC and HGA components in the lung and spleen. Immunohistochemically, positive staining of p53 protein was detected on both solid-type ACC and HGA cells, but cyclin D1 and HER2/neu was only seen in HGA cells. Solid-type ACC cells were immunoreactive for CD117 (c-kit), but lower-grade ACC and HGA cells were negative. This case suggests that the overexpression of CD117, p53 protein, cyclin D1, and HER2/neu might be involved in the progression from lower-grade ACC to solid-type ACC and HGA.

Topics: Actins; Adenocarcinoma; Aged; Carcinoma; Carcinoma, Adenoid Cystic; Disease Progression; Fatal Outcome; Humans; Immunohistochemistry; Keratins; Liver Neoplasms; Lung Neoplasms; Male; Maxillary Sinus Neoplasms; Muscle, Smooth; S100 Proteins; Tumor Suppressor Protein p53

Pulmonary epithelium is known to undergo a preneoplastic process prior to the development of lung carcinoma. Squamous dysplasia and atypical adenomatous hyperplasia have been identified and classified as preinvasive lesions of squamous cell carcinoma and peripheral pulmonary adenocarcinoma, respectively. However, these commonly recognized preinvasive lesions do not completely explain the development of all histological types of lung carcinoma. By examining 114 resection lung specimens, we concluded that there are four histological patterns of bronchial epithelial dysplasia based on morphological features (basal cell dysplasia, columnar cell dysplasia, bronchial epithelial dysplasia with transitional differentiation, and squamous dysplasia). The histological patterns were further characterized by immunohistochemistry. Basal cell dysplasia was focally positive for cytokeratin (CK) 17 and 10/13; columnar cell dysplasia was generally positive for CK7, 8, and 18; bronchial epithelial dysplasia with transitional differentiation had a heterogeneous immunoprofile, while squamous dysplasia was positive for CK10/13 and focally positive for CK17. Various degrees of abnormal expression of p53 and Ki-67 were found in the different types of bronchial epithelial dysplasia. The cases were divided into three groups based on degree and extent of bronchial epithelial dysplasia. By Crosstabs McNemar test, the Mann-Whitney U-test (for two independent groups), the Kruskal-Wallis one-way nonparametric ANOVA (for >2 independent groups) and Spearman correlation analysis, the degree and extent of bronchial epithelial dysplasia was shown to be positively correlated with the incidence of bronchogenic carcinoma and multifocal primary lung carcinoma (P<0.05). These findings indicated the following: (1) bronchial epithelium can develop various patterns of dysplasia with abnormal/ambiguous cell differentiation and abnormal expressions of p53 and Ki-67. Thus, these bronchial epithelial dysplastic lesions may represent a preneoplastic process. (2) The degree of bronchial epithelial dysplasia may significantly predispose individuals to bronchogenic carcinoma and multifocal primary lung carcinoma.

Topics: Adult; Aged; Bronchial Neoplasms; Carcinoma, Bronchogenic; Cell Differentiation; Epithelial Cells; Female; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Lung Neoplasms; Male; Middle Aged; Precancerous Conditions; Respiratory Mucosa; Tumor Suppressor Protein p53

As both mesotheliomas and squamous carcinomas can present a wide variety of morphological patterns, they can on occasion be confused. Recently, some groups of investigators have called attention to the difficulties that sometimes exist in distinguishing between these malignancies and the need to define a panel of markers that can assist in reaching the correct diagnosis. The aim of the present study is to compare the value of the various immunohistochemical markers currently available for the diagnosis of mesothelioma and squamous carcinoma of the lung. A total of 30 epithelioid pleural mesotheliomas exhibiting a solid or predominantly solid pattern, and 30 nonkeratinizing squamous carcinomas of the lung were investigated for the expression of the following markers: podoplanin, calretinin, mesothelin, WT1, keratin 5/6, keratin 7, p63, carcinoembryonic antigen (CEA), MOC-31, Ber-EP4, B72.3, BG-8 (Lewis(y)), leu-M1 (CD15), and thyroid transcription factor-1 (TTF-1). All 30 (100%) of the mesotheliomas reacted for calretinin, mesothelin and keratin 7, 93% each for podoplanin, WT1 and keratin 5/6, 13% for Ber-EP4, 7% each for p63, MOC-31 and BG-8, and 0% for B72.3, CEA, leu-M1 and TTF-1. All 30 (100%) of the squamous carcinomas were positive for p63 and keratin 5/6, 97% for MOC-31, 87% for Ber-EP4, 80% for BG-8, 77% for CEA, 57% for keratin 7, 40% for calretinin and B72.3, 30% for leu-M1, 27% for mesothelin, 15% for podoplanin, and 0% for WT 1 and TTF-1. After analyzing the results, it is concluded that from a practical point-of-view, a combination of two positive mesothelioma markers (WT1 and calretinin or mesothelin) with two negative mesothelioma markers (p63 and MOC-31) would allow the differential diagnosis to be established between epithelioid mesotheliomas and squamous carcinomas of the lung in nearly all instances.

Topics: Biomarkers, Tumor; Calbindin 2; Carcinoma, Squamous Cell; Diagnosis, Differential; Epithelioid Cells; GPI-Linked Proteins; Humans; Immunohistochemistry; Keratin-5; Keratin-6; Keratins; Lung Neoplasms; Membrane Glycoproteins; Mesothelin; Mesothelioma; S100 Calcium Binding Protein G; WT1 Proteins

It is important to discriminate between primary and secondary lung cancer. However, often, the discriminating diagnosis of primary lung acinar adenocarcinoma and lung metastasis of colorectal cancer based on morphological and pathological findings is difficult. The purpose of this study was to evaluate the clinical usefulness of immunohistochemistry of beta-catenin, cytokeratin (CK) 7, and CK20 for the discriminating diagnosis of lung cancer.. We performed immunohistochemistry of beta-catenin, CK7, and CK20 in 19 lung metastasis of colorectal cancer samples, 10 corresponding primary colorectal cancer samples and 11 primary lung acinar adenocarcinoma samples and compared the levels of accuracy of the discriminating diagnosis by using antibodies against these antigens.. Positive staining of beta-catenin was observed in all the lung metastasis of colorectal cancer samples as well as in the primary colorectal cancer samples but in none of the primary lung acinar adenocarcinoma samples. Positive staining of CK7 was observed in 90.9% of the primary lung acinar adenocarcinoma samples and in 5.3% of the lung metastasis of colorectal cancer samples, but in none of the primary colorectal cancer samples. Positive staining of CK20 was observed in all the primary colorectal cancer samples and in 84.2% of the lung metastasis of colorectal cancer samples, but in none of the primary lung acinar adenocarcinoma samples.. Combined immunohistochemistry of beta-catenin, CK7, and CK20 is useful for making a discriminating diagnosis between lung metastasis of colorectal cancer and primary lung acinar adenocarcinoma. This method will enable accurate diagnosis of a lung tumor and will be useful for selecting appropriate therapeutic strategies, including chemotherapeutic agents and operation methods.

Topics: Adenocarcinoma; beta Catenin; Biomarkers, Tumor; Colorectal Neoplasms; Diagnosis, Differential; Gene Expression Profiling; Humans; Immunohistochemistry; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Retrospective Studies; Sensitivity and Specificity

Lung cancer, especially adenocarcinoma and large cell carcinoma, tends to spread to the pleural cavities. Once an effusion develops, the prognosis is generally dismal. Immunocytochemistry is frequently applied to effusions for diagnostic purposes, but the prognostic value of markers in malignant effusions have thus far attracted less attention. Dakopatts CK 5/6 antibody was applied to ethanol-fixed fresh cytospin preparations from malignant pleural effusions originating from 18 patients (11 men and 7 women) with a previously or later verified non-small cell lung carcinoma (NSCLC). In three cases, CK5/6 reactivity was found in part of the malignant population, whereas 10 cases showed reactivity in most tumor cells. The lack of reactivity in malignant cells was only seen in five effusions. Females showed significantly lower reactivity rates, with all negative effusions coming from female patients, whereas 9/10 effusions with reactivity in most malignant cells originated from males. CK5/6 reactivity was significantly correlated to survival, with a median survival time of 18 days for patients with CK5/6-negative tumors, and 212 days for those with positive tumors. The strong relationship between CK5/6 reactivity and survival, and the observed gender difference, warrants larger studies aimed at the clinical utility of CK5/6 as a prognostic marker in metastatic NSCLC, the possible functional role of CK5/6 in cell adhesion in advanced NSCLC and its possible hormonal control.

Topics: Aged; Aged, 80 and over; Antibodies; Carcinoma, Non-Small-Cell Lung; Female; Humans; Immunohistochemistry; Keratin-5; Keratin-6; Keratins; Lung Neoplasms; Male; Middle Aged; Pleural Effusion; Predictive Value of Tests; Prognosis; Survival Analysis

Topics: Adult; Biopsy, Fine-Needle; Carcinoid Tumor; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Mucin-1; Neoplasm Metastasis; Pulmonary Sclerosing Hemangioma; Radiography; Vimentin

Topics: Adult; Chromosomes, Human, Pair 18; Chromosomes, Human, X; Humans; Immunohistochemistry; Karyotyping; Keratins; Lung; Lung Neoplasms; Male; Neoplasms, Radiation-Induced; Radiotherapy; Sarcoma, Synovial; Translocation, Genetic; Wilms Tumor

A 63-year-old man who had undergone resection of colon cancer 15 years previously was found to have a right hilar mass on chest X-ray, and an elevated serum carcinoembryonic antigen level. The hilar lymph nodes were resected with the right upper lobe, and the initial diagnosis was colon cancer metastasis to the right hilar lymph nodes. Although the resection was incomplete, and no additional treatment was given, the patient remained free of recurrence for 10 years. This prompted us to reconsider our diagnosis using immunohistochemistry. The resected lymph nodes were found to be positive for thyroid transcription factor 1 (TTF-1) and cytokeratin 7, and negative for surfactant apoprotein (SAP), cytokeratin 20, and napsin A. The neuroendocrine markers and thyroglobulin were also negative. These findings led us to diagnose T0N1 lung cancer. There are reports of patients with clinical T0N1,2 lung cancer having exceptionally good prognoses despite noncurative treatment; however, to our knowledge, this is the first case of a patient with T0N1 lung cancer diagnosed by immunohistochemistry, with a good prognosis despite incomplete resection. In this case, TTF-1 and cytokeratin staining was particularly helpful in the differential diagnosis.

Topics: Adenocarcinoma; Apoproteins; Aspartic Acid Endopeptidases; Colonic Neoplasms; Humans; Immunohistochemistry; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Lymph Nodes; Male; Middle Aged; Neoplasm Staging; Nuclear Proteins; Prognosis; Pulmonary Surfactant-Associated Proteins; Thyroid Nuclear Factor 1; Time Factors; Transcription Factors

The central type and peripheral type squamous cell carcinoma (SCC) of the lung have different clinicopathological characteristics, but, little is known about their biological characteristics. We investigated differences between the properties and phenotypes of peripheral-type (P-type) and central-type (C-type) SCC by performing an immunohistochemical analysis of each type by tissue microarray analysis with a large panel of antibodies. To examine strictly, we selected 20 P-type SCCs that were pathological stage T1 and limited to more peripherally than the fifth bronchial bifurcation, and 21 C-type SCCs that were pathological stage T1 and limited to a lobar bronchus. The results of the clinicopathological study showed that the patients with P-type SCC were significantly older than the patients with C-type SCC and that squamous metaplasia was predominant in C-type SCC than in P-type SCC. The 36 antibodies revealed different expression patterns of cytokeratin 7 (CK 7) and cytokeratin 19 (CK 19) between C-type and P-type SCC. CK 7 expression was more predominant in P-type SCC than in C-type SCC, and CK 19 expression was more predominant in C-type SCC than in P-type SCC. These results suggest that C-type and P-type SCC have different clinicopathological and biological features.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Tissue Array Analysis

This study was designed to examine the relationship between the presence of occult neoplastic cells (ONCs) in lymph nodes (LNs) and survival in 238 patients with stage I/II LN-negative cancer of the breast, lung, or stomach. In addition, immunohistochemistry for TS and DPD was used to compare the 5-FU sensitivity of the primary tumor in ONC (+) patients. The 5-year relapse-free survival (RFS) rate of 215 ONC (-) patients and 23 ONC (+) patients was 95.2 and 82.6%, respectively (p=0.0107). The 5-year overall survival (OS) rate of the ONC (-) and (+) patients was 97.4 and 77.4%, respectively (p=0.0000). The 6 ONC (+) patients with recurrence showed high and low TS expression in 33.3% (2/6) and 66.7% (4/6), respectively, while high and low DPD expression was observed in 16.7% (1/6) and 83.3% (5/6), respectively. In the 17 ONC (+) patients without recurrence, the corresponding values were 64.7% (11/17), 35.3% (6/17), 29.4% (5/17), and 70.6% (12/17). Patients with a combination of high TS and low DPD expression accounted for 33.3% (2/6) of the ONC (+) patients with recurrence and 52.9% (9/17) of those without recurrence, showing no significant difference between the two groups. These results suggest that ONCs are associated with a lower survival rate and that ONC (+) patients are unlikely to respond to 5-FU+LV therapy.

Topics: Antimetabolites, Antineoplastic; Breast Neoplasms; Dihydrouracil Dehydrogenase (NADP); Female; Fluorouracil; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Neoplasm Recurrence, Local; Neoplasm Staging; Stomach Neoplasms; Survival Analysis; Thymidylate Synthase

Detection of micrometastatic disease is an interesting area in non-small cell lung cancer (NSCLC). We conducted a study to determine whether the detection of mediastinal lymph node spread by immunohistochemical (IHC) analysis offers some prognosis with respect to patients' disease-free survival or not.. Between 1997 and 2003, twenty-one early stage lung cancer patients underwent complete resection with mediastinoscopy and systemic nodal dissection. Four hundred and twenty-six paraffin-embedded lymph node sections from 21 patients were analyzed. Epithelial specific-antigen Ab-9 and Keratin-Pan Ab-1 were used as IHC marker.. Based on nodal spread four of the 21 patients (19.04%) were up-staged after IHC analysis. Two patients with stage IB (T2N0) up-staged to stage IIIA (T2N2); two patients staged as IIB (T2N1) up-staged to IIIA (T2N2). Statistical analysis showed that the lymphatic dissemination detected with IHC analysis was associated with reduced disease-free survival (DFS) (p = 0.002).. Our study provides some indication that patients with lymphatic micrometastasis have a reduced DFS. Before creating a new TNM staging system, more information is needed to understand the prognostic impact of micrometastatic dissemination.

Topics: Antibodies, Neoplasm; Biomarkers, Tumor; Carcinoma; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Retrospective Studies

We evaluated the sensitivity and specificity of 10 monoclonal and two polyclonal antibodies for distinguishing epithelioid mesothelioma from adenocarcinoma (AdCA) using immunohistochemistry (IHC). The antibodies were directed against the mesothelial-associated antigens mesothelin, calretinin, cytokeratin 5, thrombomodulin, Wilms' tumor-1 (WT-1) gene product and HBME-1, and the nonmesothelial antigens Lewis-Y blood group (antibody BG8), MOC-31, BerEp4, CD15, and carcinoembryonic antigen (CEA) family. The 133 tumors evaluated included 65 malignant epithelioid mesotheliomas, 22 lung AdCAs, 27 ovarian serous carcinomas, 24 breast carcinomas, and five gastric carcinomas. Diagnoses were based on clinical, histologic, ultrastructural, and/or IHC findings. Calretinin had the best sensitivity for mesothelioma (95%), followed by HBME-1 (84%), WT-1 (78%), cytokeratin 5 (76%), mesothelin (75%), and vimentin and thrombomodulin (68%). Thrombomodulin had the best specificity for mesothelioma (92%), followed by cytokeratin 5 (89%), calretinin (87%) vimentin (84%), and HBME-1 (45%). When ovarian carcinomas were excluded from the analysis, the specificity of mesothelin and WT-1 for the diagnosis of mesothelioma increased to 90 and 81%, respectively. The sensitivity of the nonmesothelial antigens for AdCA was organ dependent, with BG8 performing best in the breast cancer group (96%), and BerEp4, BG8, MOC-31 performing best in the lung cancer group (100%). The specificity of the nonmesothelial antigens for AdCA was 98% for BG8 and CEA, 97% for CD15, 95% for BerEp4, and 87% for MOC-31. A novel statistical analysis technique employing logic regression analysis identified a three-antibody immunohistochemical panel including calretinin, BG8, and MOC-31, which provided over 96% sensitivity and specificity for distinguishing epithelioid mesothelioma from AdCA.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antibody Specificity; Biomarkers, Tumor; Breast Neoplasms; Calbindin 2; Carcinoembryonic Antigen; Diagnosis, Differential; Female; GPI-Linked Proteins; Humans; Immunohistochemistry; Keratin-5; Keratins; Lung Neoplasms; Membrane Glycoproteins; Mesothelin; Mesothelioma; Multivariate Analysis; Ovarian Neoplasms; S100 Calcium Binding Protein G; Sensitivity and Specificity; Stomach Neoplasms; Thrombomodulin; WT1 Proteins

Thyroid transcription factor-1 (TTF-1), and cytokeratin 7 (CK7) and cytokeratin 20 (CK20) have recently been reported to be useful to distinguish between primary and metastatic lung adenocarcinoma. The aim of this study was to determine the usefulness of the staining patterns of pulmonary adenocarcinoma with antibodies to TTF-1, CK7, and CK20 in differentiating primary from metastatic pulmonary adenocarcinoma. Of the 66 lung adenocarcinoma specimens that were enrolled in our study, there were 40 primary lung adenocarcinomas, 12 metastatic adenocarcinomas from breast, 13 metastatic adenocarcinomas from colon, and 1 metastatic adenocarcinoma from stomach. The expression of TTF-1, CK7, and CK20 was assessed by immunohistochemistry. We found that 73% of primary lung adenocarcinomas expressed TTF-1, whereas all nonpulmonary adenocarcinomas lacked TTF-1 staining. CK7 expression was significantly more frequent in adenocarcinomas of pulmonary and breast origin than gastrointestinal (GI) origin (p < 0.001). In contrast, CK20 expression was significantly more prevalent in adenocarcinoma that originated in the GI tract than that of pulmonary or breast origin (p < 0.001). A combination of TTF-1+CK7+CK20- was highly significantly associated with primary adenocarcinoma of lung (vs GI tract, p < 0.001; vs breast, p < 0.001). A combination of TTF-1-CKCK20+ was highly significantly associated with adenocarcinoma of GI origin (vs lung, p < 0.001; vs breast, p < 0.001). Our study has confirmed that expression of CK7, CK20, and TTF-1 is a useful immunohistochemical marker for diagnosis of lung tumors and for differential diagnosis of primary pulmonary adenocarcinomas from extrapulmonary adenocarcinomas metastatic to the lung. Application of this panel of antibodies might be expected to increase the accuracy of diagnosis.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Female; Humans; Immunohistochemistry; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

We report a rare case of unknown primary carcinoma. A 36-year-old man was admitted to the hospital because of a chest wall tumor. Serum carcinoembryonic antigen level was 160 ng/ml. The resected chest wall tumor was pathologically diagnosed as metastatic adenocarcinoma, showing positive immunoreactivity for cytokeratin 7 and negative immunoreactivity for cytokeratin 20, suggesting lung origin. Serum carcinoembryonic antigen level returned to normal limits. Twenty-one months later, a chest X-ray showed a nodular lesion in the left upper lobe and serum carcinoembryonic antigen level increased to 12.3 ng/ml. Left upper lobectomy was performed 23 months after chest wall resection. The resected tumor was pathologically diagnosed as primary lung adenocarcinoma, showing the same immunoreactivity as in the chest wall tumor. The combination of immunohistochemistry for cytokeratin 7 and 20 appeared to be a useful tool in determining the site of origin and helpful for premortem diagnosis of the origin of unknown primary carcinoma.

Topics: Adenocarcinoma; Adult; Humans; Immunohistochemistry; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Neoplasms, Unknown Primary; Thoracic Neoplasms; Thoracic Wall

The nonangiogenic lung tumour is characterized by neoplastic cells co-opting the pre-existent vasculature and filling the alveoli space. 3-dimensional reconstruction of the tumour reveals that this particular tumour progresses without neovascularization and there is no major destruction of the lung's architectural integrity.

Topics: Antigens, CD34; Carcinoma, Non-Small-Cell Lung; Disease Progression; Humans; Imaging, Three-Dimensional; Immunohistochemistry; Keratins; Lung Neoplasms; Neovascularization, Pathologic; Staining and Labeling

We assessed the usefulness of several immunohistochemical stains in distinguishing these two neoplasms, including cytokeratin 7, cytokeratin 20 (CK20), neuron-specific enolase, chromogranin, synaptophysin, neurofilaments (NF), thyroid-transcription factor-1 (TTF-1), CD56 antigen, S-100 protein, vimentin, c-erbB-2 oncoprotein, and CD117 antigen. All 13 cases of Merkel cell carcinoma evaluated were positive for CK20, and negative for TTF-1. Twelve of 13 Merkel cell carcinoma cases were positive for NF. Eleven of 13 cases of small cell lung carcinoma were positive for TTF-1. All small cell lung carcinoma cases were negative for NF, and all but one were negative for CK20. In terms of the remaining antigens, there were no differences of significance between the two neoplasms. These findings suggest that a set of three immunohistochemical stains, including CK20, NF, and TTF-1, is useful in affording a distinction between Merkel cell carcinoma and small cell lung carcinoma.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Merkel Cell; Carcinoma, Small Cell; CD56 Antigen; Chromogranins; Female; Humans; Immunohistochemistry; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Neurofilament Proteins; Nuclear Proteins; Phosphopyruvate Hydratase; Proto-Oncogene Proteins c-kit; Receptor, ErbB-2; S100 Proteins; Skin Neoplasms; Synaptophysin; Thyroid Gland; Thyroid Nuclear Factor 1; Transcription Factors; Vimentin

Cytokines are potential new serum markers, especially desirable for malignancies with poor prognosis like non-small cell lung cancer (NSCLC).. Cytokines, tumor necrosis factor alpha (TNFalpha), interleukin (IL)-6 and IL-8, soluble TNF (sTNF) RI, sTNF RII, soluble IL-2 receptor-alpha, IL-1 receptor antagonist (IL-1ra), IL-10, vascular endothelial growth factor, basic fibroblast growth factor, and macrophage (M-CSF) and granulocyte colony-stimulating factor, as well as tumor markers - carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC) and CYFRA 21.1 - were assessed in the sera of 103 untreated NSCLC patients, and these cytokines and tumor markers were referred to clinical parameters of the disease and to the overall survival of patients evaluated during a 6-year follow-up.. Most of the factors analyzed were found to be elevated in the sera of NSCLC patients, and increases in IL-6, IL-8 and sTNF RI were noted in the greatest proportion of stage I patients. Most cytokine/cytokine receptor levels revealed higher sensitivity than the standard tumor markers; IL-6 and IL-1ra levels were significantly different in patients with squamous cell versus adenocarcinoma; IL-6 and IL-10 were related to the tumor size, while IL-6 and M-CSF levels significantly increased with disease progression. A significant prognostic value of pretreatment serum M-CSF and CEA levels in NSCLC patients has been shown, but only M-CSF proved to be an independent prognostic factor.. Increased pretreatment serum M-CSF level is a significant independent predictor of poor survival in patients with NSCLC.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Cytokines; Female; Fibroblast Growth Factor 2; Humans; Interleukins; Keratin-19; Keratins; Lung Neoplasms; Macrophage Colony-Stimulating Factor; Male; Middle Aged; Predictive Value of Tests; Prognosis; Receptors, Cytokine; Receptors, Interleukin; Receptors, Tumor Necrosis Factor; Risk Factors; ROC Curve; Serpins; Time Factors; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

In this study, we sought to validate the importance of p63, CK5/CK6, CK7, and surfactant-A (SP-A) to classify 42 non-small cell lung cancers in autopsy and surgical resection specimens and to study the usefulness of these markers in distinguishing between squamous cell carcinomas and adenocarcinomas because of their different implications regarding treatment and prognosis. All adenocarcinoma cases were negative for p63; 9 (56.2%) of 16 were CK5/CK6 positive, 16 (94.1%) of 17 were CK7 positive, and 4 (26.6%) of 15 were SP-A positive. In squamous cell carcinoma, 1 case was CK7 and SP-A positive and 14 (77.8%) of 18 were p63 positive. The latter appears to be useful in differentiating squamous cell carcinoma from adenocarcinoma of the lung in small biopsies without keratinization or glandular differentiation; thus, for advanced-stage cases, where there is no possibility of surgical resection and the treatment of choice is radiotherapy plus chemotherapy, we would be able to differentiate between the two histological types, establishing then a different therapy.

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratin-5; Keratin-6; Keratin-7; Keratins; Lung Neoplasms; Membrane Proteins; Pulmonary Surfactant-Associated Protein A

Topics: Actins; Antigens, Neoplasm; Desmin; Diagnosis, Differential; Female; Humans; Hyperplasia; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Lymphangioleiomyomatosis; Melanoma-Specific Antigens; Middle Aged; Muscle, Smooth; Neoplasm Proteins; Nuclear Proteins; Receptors, Estrogen; Receptors, Progesterone; Thyroid Nuclear Factor 1; Transcription Factors; Tuberous Sclerosis

To elucidate additional phenotypic differences between large cell neuroendocrine carcinoma (LCNEC) and small cell lung carcinoma (SCLC), we performed tissue microarray (TMA) analysis of surgically resected LCNEC and SCLC specimens. Immunostaining with 48 antibodies was scored based on staining intensity and the percentage of cells that stained positively. Four proteins were identified as significantly expressed in LCNEC as compared with SCLC: cytokeratin (CK)7, 113 vs 49 (P < .0301); CK18, 171 vs 60 (P < .0008); E-cadherin, 77 vs 9 (P < .0073); and beta-catenin, 191 vs 120 (P < .0286). Immunostaining of cross-sections containing LCNEC and SCLC components revealed significant expression of CK7, CK18 and beta-catenin in the LCNEC component compared with the SCLC component in 2 of 3 cases. Our results indicate that significant expression of CK7, CK18, E-cadherin, and beta-catenin is more characteristic of LCNEC than of SCLC, and these findings provide further support that these tumor types are separate entities morphologically and immunophenotypically, if not biologically.

Topics: beta Catenin; Biomarkers, Tumor; Cadherins; Carcinoma, Large Cell; Carcinoma, Neuroendocrine; Carcinoma, Small Cell; Diagnosis, Differential; Humans; Immunohistochemistry; Keratin-7; Keratins; Lung Neoplasms; Neoplasm Proteins; Retrospective Studies; Tissue Array Analysis

The ability to detect occult systemic metastases in patients with operable NSCLC could have a significant impact on the management of the disease. The aim of the study was to detect occult micrometastatic tumor cells in bone marrow in patients with resectable NSCLC.. A total of 35 patients (29 men, 6 women), age between 47 and 78 (mean 61.6) were included in the study. In each of the patients bone marrow aspirates from the ribs were sampled during surgery. Both the tumor and the bone marrow aspirate were examined histologically and immunocytochemically with the cytokeratin: AE1/AE3, CAM 5,2, CK-7, CK-18. The presence of grow factors CD 31 and CD 34 were examined as well.. No evidence of micrometastases or tumor cells in bone marrow was found in histological examination. Cytokeratin positive (CAM 5,2 +) cells were detected in 33 cases (94.23%) of the tumors and in 21 cases (60.00%) of bone marrow samples. The statistically significant correlation between the presence of CAM 5,2 in tumors and bone marrow was found (p = 0.049). Cytokeratin positive cells were detected in all the 35 tumors (AE1/AE3), in 20 tumors--57.14% (CK-7) and in 23 tumors--65.71% (CK-18). Cytokeratin positive cells (CK-7) were detected in bone marrow sample in one patient only.. Immunocytochemical examination with the use of cytokeratin CAM 5,2 is of use to detect occult micrometastatic tumor cells in bone marrow in NSCLC patients. However, no correlations were found between the presence of cytokeratin CAM 5,2 in bone marrow or tumor and patients' age, sex and the histological type of NSCLC its degree of malignancy and stage.

Topics: Aged; Biomarkers, Tumor; Biopsy, Needle; Bone Marrow; Carcinoma, Non-Small-Cell Lung; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Neoplastic Cells, Circulating; Prognosis

Our previous proteomics study on human hepatocellular carcinoma (HCC) cell strains revealed that cytokeratin 19 (CK19) was expressed in cells with high metastasis potential; we further studied serum CK19 fragment CYFRA 21-1 level in HCC patients and nude mice model of HCC metastasis. HCC cell line HCCLM3 was injected subcutaneously into 30 nude mice which were then randomized into 6 groups of 5 mice each. The murine serum CYFRA 21-1 and pulmonary metastases were determined 2, 3, 4, 5, 6, and 7 weeks after injection. Serum CYFRA 21-1 levels of 101 normal controls and 108 HCC patients were also determined. In nude mice model, CYFRA 21-1 level increased significantly when pulmonary metastases occurred. Among 108 HCC patients, 24 (22.2%) had increased serum CYFRA 21-1 level. The presence of portal vein tumor emboli was significantly higher in CYFRA 21-1 increased cases (33.3%, 6/24) than in CYFRA 21-1 normal cases (6.0%, 5/84) (x2=7.403, P < 0.01). In addition, the percentage of TNM stage III/IV tumor was significantly higher in CYFRA 21-1 increased patients (54.2%, 13/24) than in CYFRA 21-1 normal cases (21.4%, 18/84) (x2=9.776, P < 0.005). These results suggest that CK19 may play an important role in HCC metastasis.

Topics: Adult; Aged; Animals; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Hepatocellular; Female; Humans; Keratin-19; Keratins; Liver Neoplasms; Lung Neoplasms; Male; Mice; Mice, Nude; Middle Aged; Neoplasm Staging; Neoplastic Cells, Circulating; Portal Vein; Random Allocation

Detection of tumor-related mRNA in blood has become a potential cancer diagnostic approach. However, the sensitivity of single-marker assays is not high enough for clinical applications. The present study was aimed to evaluate the efficacy of a multimarker panel for molecular diagnosis of non-small cell lung cancer (NSCLC).. Carcinoembryonic antigen (CEA), cytokeratin 19 (CK-19), c-met and heterogeneous nuclear ribonucleoprotein (hnRNP) B1 mRNAs were quantified by quantitative real-time reverse transcriptase polymerase chain reaction in 34 tumor tissues and 69 peripheral blood samples of NSCLC patients.. All four markers displayed high overexpression rates (range 82.3-97.1%) in NSCLC tumors. When used as single markers in blood for NSCLC diagnosis, CEA, CK-19, c-met and hnRNP B1 could only reach sensitivities of 52.2, 50.7, 42 and 17.4%, respectively. However, the sensitivity was enhanced up to 85.5% when CEA, CK-19 and c-met were combined in a 3-marker panel. Moreover, the expression of c-met and hnRNP B1 in blood was significantly correlated with patients' pathological stages.. The combined detection of CEA, CK-19 and c-met mRNAs in blood provided a valuable tool for molecular diagnosis of NSCLC. In addition, our results also suggested that hnRNP B1 was not a valuable diagnostic marker but a potential prognostic marker for NSCLC.

Topics: Actins; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Gene Expression Regulation, Neoplastic; Heterogeneous-Nuclear Ribonucleoprotein Group A-B; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Polymerase Chain Reaction; Proto-Oncogene Proteins c-met; RNA, Messenger; Up-Regulation

Non-small cell lung cancer (NSCLC) is derived from epithelial cells and accounts for approximately 80% of all lung cancers. Cytokeratins are specific for epithelial cells and during malignant transformation the cytokeratin profile usually remains constant. Angiogenesis is the formation of new blood vessels out of the existing vascular bed. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are potent circulating angiogenic factors. The aim of the present study was to determine if increased levels of a new cytokeratin assay (MonoTotal, which in comparison with TPAcyk detects not only fragments of cytokeratins 8 and 18 but also of cytokeratin 19) is correlated with circulating angiogenic factors (VEGF and bFGF) and the secondary aim was to investigate if increased levels of these circulating markers are associated with survival. In the present study, a total of 45 NSCLC patients (26 patients stage IIIa and 19 patients stage IIIb) receiving only curatively intended treatment for advanced NSCLC were included. These patients donated a total of 291 serum samples during follow-up which was investigated for the presence of MonoTotal, VEGF and bFGF. MonoTotal was statistically significantly correlated with bFGF (R=0.26, p=0.00049) and VEGF (R=0.26, p=0.00007). From the time of histological diagnosis until time of death, MonoTotal increased by 603 U/l (p<0.0001). VEGF increased by 430 pg/ml (p=0.0004) whereas the corresponding value for bFGF was 5.93 pg/ml (p=0.018). MonoTotal, a newly developed commercial cytokeratin assay, seems to be a potentially very interesting serum marker that, in conjunction with other clinical data, might be used for monitoring of patients with NSCLC.

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Fibroblast Growth Factor 2; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Risk; Survival Analysis; Vascular Endothelial Growth Factor A

We encountered a male patient aged 64 with pulmonary mucinous carcinoma in whom a diagnosis of pulmonary metastasis from early rectal cancer with submucosal invasion was made based on an immunohistochemical examination. A rectal cancer was detected together with a mass in the lung. The mass in the lung was consistent with mucinous adenocarcinoma, whereas the invasion of rectal cancer was confined to the submucosa; thus, distant metastases appeared unlikely. These lesions were assessed using immunohistochemical staining for cytokeratin and thyroid transcription factor-1, which failed to make a definite diagnosis. A further assessment was made by staining for villin. Both neoplasms were positive for this protein, demonstrating a common brush-border pattern. A lung metastasis from rectal cancer with submucosal invasion was diagnosed. Villin is considered useful for detecting primary neoplastic lesions based not only on its specificity but also on its staining pattern, which is different from that of other proteins.

Topics: Adenocarcinoma, Mucinous; Biomarkers, Tumor; Fatal Outcome; Humans; Immunohistochemistry; Intestinal Mucosa; Keratins; Lung Neoplasms; Male; Microfilament Proteins; Middle Aged; Neoplasm Invasiveness; Nuclear Proteins; Rectal Neoplasms; Thyroid Nuclear Factor 1; Transcription Factors

The authors assessed the predictive and prognostic role of decline in the serum levels of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA 21-1) during chemotherapy in patients with advanced nonsmall cell lung cancer (NSCLC).. Changes in serum levels of CEA and CYFRA 21-1 during first-line, conventional chemotherapy were studied prospectively with an immunometric assay at baseline and every 2 courses in 117 patients with advanced NSCLC. Data were correlated with radiologic objective response (OR) and survival.. One hundred seven patients were evaluable for radiologic and serologic response assessment after 2 chemotherapy courses. The radiologic OR rate was 44% overall. The CEA and CYFRA 21-1 responses (> or =20% reduction over baseline level; assessed after the second course of chemotherapy) were 38% and 61%, respectively. Statistically significant correlations were observed between CEA and CYFRA 21-1 responses and OR (P = .01 and P = .004, respectively). The median survival from response assessment (landmark analysis) was 9 months. In a univariate analysis, disease stage, performance status, baseline lactate dehydrogenase level (LDH), OR, CEA response, and CYFRA 21-1 response were correlated significantly with survival. In particular, the median survival was 13 months for patients who had a CEA response and 11 months for patients who had a CYFRA 21-1 response compared with 8 months and 6 months for patients who did not respond, respectively. In a multivariate analysis, performance status (P = .005), baseline LDH level (P = .02), CEA response (P = .03) and CYFRA 21-1 response (P = .01) were confirmed as independent prognostic factors for survival.. CEA and CYFRA 21-1 responses appeared to be reliable surrogate markers of chemotherapy efficacy in patients with advanced NSCLC.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Prognosis; Prospective Studies; Treatment Outcome

Using current diagnostic criteria, this work summarizes the microscopic review of 16 proliferative squamous lesions, previously diagnosed as cystic keratinizing squamous cell carcinoma, in the lungs of rats from a 2-year inhalation study with pigment-grade titanium dioxide particles.. In the aftermath of two international pathology workshops designed, in part, to establish histological criteria for classifying pulmonary keratin lesions, these lesions were evaluated by four pathologists using current diagnostic criteria.. Unanimous agreement was reached as to the diagnosis of each of the lesions. Two of the lesions were diagnosed as squamous metaplasia and one as a poorly keratinizing squamous cell carcinoma. The remaining 13 lesions were diagnosed as non-neoplastic pulmonary keratin cysts.. These keratin cysts are a species-specific lesion that is unique to the rat lung under conditions of particle overload exposure.

Topics: Animals; Cysts; Dose-Response Relationship, Drug; Dust; Environmental Exposure; Female; Inhalation Exposure; Keratins; Lung Diseases; Lung Neoplasms; Male; Neoplasms, Squamous Cell; Pulmonary Alveoli; Rats; Species Specificity; Titanium

We developed and validated a novel method for quantifying protein expression of cancer tumors in an accurate, sensitive, and high throughput format. This technique integrates quantum dots, tissue microarray, optical spectroscopy, and algorithm design for analysis of tumor biopsies. The integration of this method for tissue analysis in the clinic bears potential impact for improving the diagnosis and treatment of cancer.

Topics: Animals; Antigens, Neoplasm; Cadherins; Cell Line, Tumor; ErbB Receptors; Fluorescence Resonance Energy Transfer; Humans; Keratins; Lung Neoplasms; Mice; Mice, SCID; Microarray Analysis; Neoplasm Transplantation; Quantum Dots; Transplantation, Heterologous

To evaluate the performance of 18F-FDG three-head tomography with coincidence imaging and serum tumor marker assays in identifying lung lesions in 104 patients with abnormal findings on chest X-ray or computer tomography.. A prospective evaluation of 18F-FDG coincidence imaging and the measurement of 3 serum markers for lung cancer ( carcinoembryonic antigen, CYFRA21-1 and neuron specific enolase) were performed within one week in 104 inpatients with suspected lung malignancy. All images were analyzed visually. It was considered positive for malignancy if the 18F-FDG uptake was increased relative to that in the adjacent lung tissue, and was focal. The serum tumor marker test was considered positive for malignancy if the serum level of at least one marker was elevated.. 66 patients were proven to have lung cancer by pathology, and 38 patients had benign lung diseases. The sensitivity, specificity, accuracy of 18F-FDG coincidence imaging and serum tumor markers in assessing lung cancers were 80. 0% , 77. 2% , 77. 9% and 56. 0% , 60. 9%, 64. 4% , respectively. 18F-FDG coincidence images in assessing lung lesions showed significantly higher sensitivity, specificity and accuracy than serum tumor markers. Four patients with lung cancer had negative findings on 18F-FDG coincidence images but showed positive serum markers.. 18F-FDG coincidence imaging is a powerful tool for evaluating patients with lung lesions suggestive of malignancy. Although the determination of serum marker levels is less accurate than 18F-FDG coincidence imaging, the combination of a positive 18F-FDG coincidence result and positive tumor markers may be helpful in improving the diagnosis of lung cancers.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Fluorodeoxyglucose F18; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Plasma Cell Granuloma, Pulmonary; Positron-Emission Tomography; Prospective Studies; Radiopharmaceuticals; Sensitivity and Specificity; Tuberculosis, Pulmonary

Accurate morphologic distinction between small cell carcinoma and poorly differentiated squamous cell carcinoma has critical therapeutic significance, but can be limited by crush artifact, tumor necrosis, limited tumor representation, and overlapping morphologic features. We evaluated a panel of antibodies for their efficacy in distinguishing between these neoplasms. Formalin-fixed paraffin-embedded tissue sections of small cell carcinomas and poorly differentiated squamous cell carcinomas underwent immunohistochemical staining with antibodies to thyroid transcription factor-1, p63, high molecular weight keratin, and p16(INK4A). Of 28 small cell carcinomas, 26 (93%) small cell carcinomas showed diffuse moderate or strong staining for thyroid transcription factor-1 with no staining for high molecular weight keratin and p63. In contrast, 27/28 (96%) poorly differentiated squamous cell carcinomas manifested opposite immunoreactivities, with diffuse moderate or strong staining for high molecular weight keratin and p63, and no or minimal staining for thyroid transcription factor-1. In two additional cases originally interpreted as small cell carcinoma, high molecular weight keratin highlighted small numbers of neoplastic large cells, leading to reclassification as combined small cell and non-small cell carcinomas. p16(INK4A) expression varied widely in poorly differentiated squamous cell carcinomas, but was consistently moderate or strong and diffuse in small cell carcinomas, and proved helpful in the two thyroid transcription factor-1-negative small cell carcinomas. This study demonstrates that a panel consisting of antibodies to thyroid transcription factor-1, p63, high molecular weight keratin, and p16(INK4A) is highly effective for distinguishing between small cell carcinoma and poorly differentiated squamous cell carcinoma. This panel also facilitates diagnosis of combined small cell and non-small cell carcinomas.

Topics: Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Differentiation; Cyclin-Dependent Kinase Inhibitor p16; Diagnosis, Differential; DNA-Binding Proteins; Genes, Tumor Suppressor; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Nuclear Proteins; Phosphoproteins; Thyroid Nuclear Factor 1; Trans-Activators; Transcription Factors; Tumor Suppressor Proteins

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) as gefitinib emerged as an accepted treatment in second- or third-line setting in NSCLC. However, clinical surrogate markers of EGFR-TKI activity in NSCLC patients remain to be identified and we studied the prognostic value of CYFRA 21-1 in this setting. Serum samples from 53 patients with NSCLC receiving gefitinib after failure of at least a platinum-containing regimen were prospectively collected from January 2002 to December 2003. Multivariate analysis demonstrated an independent negative impact on survival for a level of CYFRA 21-1 higher than 3.5 ng ml(-1) (HR=2.45, 95% CI 1.13-5.29; P=0.02). In conclusion, CYFRA 21-1 is a tool available to predict the survival of NSCLC patients receiving gefitinib as third-line therapy in an independent manner. In case of a CYFRA 21-1 level higher than 3.5 ng ml(-1), treatment with gefitinib needs further evaluation giving its relative poor effect on survival.

Topics: Aged; Antigens, Neoplasm; Antineoplastic Agents; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Gefitinib; Humans; Intracellular Signaling Peptides and Proteins; Keratin-19; Keratins; Lung Neoplasms; Male; Prognosis; Quinazolines

Six cases of an unusual variant of primary pulmonary adenocarcinoma resembling colorectal and sinonasal adenocarcinoma are presented. Pulmonary intestinal-type adenocarcinoma occurs in elderly Caucasians and is associated with a histology characteristic of colorectal/enteric adenocarcinoma: a garland-like architecture with a 'gland in gland' periphery, central 'dirty' necrosis, and elongated stratified columnar cells, lacking significant goblet or signet ring differentiation. While a resemblance to intestinal adenocarcinoma by light microscopy is present, immunohistochemical studies comparing these carcinomas with metastatic colorectal adenocarcinoma clearly show a respiratory phenotype with the neoplastic cells expressing thyroid transcription factor-1 and cytokeratin 7 to the exclusion of cytokeratin 20, and failing to express CDX-2. Stains for a variety of epithelial mucins (MUC1, MUC2, MUC5AC) also support this observation. The differential diagnosis with other pulmonary adenocarcinomas, especially those with mucinous differentiation, is discussed.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; CDX2 Transcription Factor; Cell Differentiation; Colonic Neoplasms; Female; Homeodomain Proteins; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Mucin 5AC; Mucin-1; Mucin-2; Mucins; Nuclear Proteins; Thyroid Nuclear Factor 1; Trans-Activators; Transcription Factors

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; CA-125 Antigen; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Disease Progression; Female; Genetic Variation; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged

This study was designed to screen occult cancer cells by CK19 mRNA detection using reverse transcriptase-polymerase chain reaction (RT-PCR) in mediastinal lymph nodes stations (MLNS) in non-small cell lung carcinoma (NSCLC). In 49 NSCLC patients free of mediastinal adenopathy on computed tomograph, 254 MLNS were evaluated by histopathology, immunohistochemistry (IHC) and RT-PCR. Of 225 non-tumoral MLNS on histopathology, 32 (14.2%) were positive by RT-PCR. IHC did not provide significant additional results. Seventeen patients were without mediastinal tumoral extension on histopathology and RT-PCR (Group 1), 16 were upgraded by RT-PCR (Group 2) and 16 pN2 on histopathology (Group 3). The two-year cancer-related death survival in Groups 1 (100%) and 2 (64.5%) was significantly different (P=0.04). The relative risk of recurrence in Group 2 compared with Group 1, evaluated by the Cox model multivariate analysis, was 5.61 (P=0.02). In conclusion, CK19 mRNA detected by RT-PCR in MLNS was significantly associated with an increased risk of rapid recurrence.

Topics: Adult; Aged; Carcinoma, Non-Small-Cell Lung; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Node Excision; Lymphatic Metastasis; Male; Mediastinal Neoplasms; Middle Aged; Multivariate Analysis; Neoplasm Recurrence, Local; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; RNA, Messenger; RNA, Neoplasm; Survival Analysis

Interstitial pneumonia (IP) may occur with the chest radiographic abnormality at the bilateral, predominantly basal and subpleural area of the lower lobe. And the incidence of lung cancer are markedly increased among patients with IP. From 1993 to 2003, 15 cases (male 14, female 1; average age 65.9-year-old), who were complicated IP, underwent surgical treatment in our hospital. Concerning detective tumor markers of lung cancers with IP, CYFRA showed very high sensitivity (92.3%) for any type of carcinoma. There were no case of perioperative death, however, 3 cases occurred severe deterioration of IP within 1 month and 6 cases died of respiratory failure. The risk factors, which aggravated IP at the postoperative period, were administration of prednisolone or immunosuppressive drugs from pre-operation and resection of the relatively non-fibrotic and non-honeycomb lobe which was affected with cancer. If it is possible to prognosticate the postoperative deterioration of IP, segmentectomy should be considered.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Diseases, Interstitial; Lung Neoplasms; Lymph Node Excision; Male; Middle Aged; Neoplasm Staging; Pneumonectomy; Prognosis; Survival Rate

Pleomorphic carcinoma of the lung (PCL) is characterized by a mixture of sarcomatoid and carcinoma components, and a poor prognosis. However, no immunophenotype of tumor markers has been characterized in PCL. To characterize the immunophenotype for CD99 in PCL, we performed an immunohistochemical evaluation of PCLs for thyroid transcription factor-1 (TTF-1), cytokeratin (CK) 7 and 20, and for CD99. CD99 was found to be expressed in both carcinomatous (47%) and sarcomatous components such as spindle cells (92%) and giant cells (57%). In the case of spindle cells, CK7 was expressed in 6 cases (46%) and TTF-1 in 2 cases (15%), whereas for giant cells CK7 was expressed in 8 cases (57%) and TTF-1 in one case (7%). However, CK20 was not expressed in either the carcinomatous or sarcomatous components in any case. Thus, CD99 was found to be widely expressed in both sarcomatous and carcinoma component in PCL. A clinicopathological analysis showed no direct correlation between the expression of CD99 and the clinical indices (stage, survival rate, invasion) of PCL.

Topics: 12E7 Antigen; Adult; Aged; Aged, 80 and over; Antigens, CD; Carcinoma; Cell Adhesion Molecules; Female; Humans; Immunohistochemistry; Immunophenotyping; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Prognosis; Sarcoma; Thyroid Nuclear Factor 1; Time Factors; Transcription Factors

Paraffin-embedded sections of various adenocarcinomas (13 colonic, 11 mucinous ovarian, 5 serous ovarian, 8 pancreatic, 6 ampullary, 12 gastric, 5 esophageal, 10 endometrial, 29 breast, and 55 lung) and 29 additional lung carcinomas (nonadenocarcinomas) were immunostained with antibodies to CDX2 protein, cytokeratin 7 (CK7), and cytokeratin 20 (CK20). The 84 lung carcinomas were also stained with antibody to thyroid transcription factor-1 (TTF-1). All colorectal and most ovarian mucinous carcinomas were strongly and diffusely immunoreactive for CDX2. Esophageal, gastric, and ampullary adenocarcinomas showed variable immunoreactivity for CDX2. All breast, nonmucinous ovarian, and most endometrial and pancreatic adenocarcinomas showed no immunoreactivity for CDX2. CK7 and CK20 expression was similar to previous reports. Ten of 84 primary lung carcinomas (12%) were immunoreactive for CDX2 expression. Of these, 5 (4 adenocarcinomas and 1 large cell carcinoma) were reactive for TTF-1. Gene expression profiling data--available for 32 of these 84 tumors--showed CDX2 gene expression in 7 of 8 (88%) CDX2 immunoreactive tumors whereas only 1 of 24 (4%) tumors negative for CDX2 immunoreactivity showed CDX2 gene expression. The authors conclude that CDX2 is a relatively specific marker for tumors with intestinal differentiation, with the caveat that its expression can be seen in primary large cell and adenocarcinomas of the lung and mucinous carcinomas of the ovary.

Topics: Adenocarcinoma; Biomarkers, Tumor; CDX2 Transcription Factor; Gastrointestinal Neoplasms; Homeodomain Proteins; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms

We report 6 cases of low-grade pulmonary mucoepidermoid carcinoma displaying a striking lymphoplasmacytic infiltrate. All six tumors had a typical pulmonary mucoepidermoid carcinoma presentation as a polypoid endobronchial mass involving the proximal bronchi. The patients were 3 females and 3 males with a mean age of 33 years (range, 5-61 years). Half of the patients were asymptomatic, while half experienced mild symptoms of pneumonia, asthma-like symptoms, or hemoptysis. No tumor-related deaths were observed, with a mean follow-up of 51 months. The tumor size ranged from 2.1 to 3.4 cm (mean, 2.9 cm). The tumors characteristically displayed an elaborate tubulocystic epithelial component composed of intermediate, epidermoid, and mucus-producing cells, and variable numbers of clear cells, multinucleated giant cells, columnar cells, and oncocytic cells. The tumors' lymphoplasmacytic infiltrate with occasional Russell bodies was sufficiently intense to raise concern of a low-grade lymphoma. All tested tumors were immunoreactive with CK7 while nonreactive with TTF-1 and CK20. Recognition of this histologic variant is important for a correct diagnosis of low-grade pulmonary mucoepidermoid carcinoma. The dense lymphoplasmacytic infiltrate is similar to that previously described in salivary glands as tumor-associated lymphoid proliferation.

Topics: Adult; Biomarkers, Tumor; Carcinoma, Mucoepidermoid; Cell Proliferation; Child, Preschool; Female; Humans; Keratin-7; Keratins; Lung Neoplasms; Lymphoid Tissue; Lymphoproliferative Disorders; Male; Middle Aged

The aim of the present study was to evaluate the usefulness of immunohistochemical markers in the differential diagnosis of pulmonary neuroendocrine tumors with particular emphasis on the preservation of immunoreactivity in areas showing crush artifacts. Specimens from 9 carcinoid tumors (CTs) and 13 small cell carcinomas (SCCs) with crush artifact were stained with antibodies to Ki-67, chromogranin A, synaptophysin, and cytokeratin. The immunoreactivity was well preserved in the crushed areas. Ki-67 was expressed in the crushed areas of all SCCs. Reactivity was diffuse or at least present in 25% of the crushed areas. In contrast, the immunoreactive areas in CTs never exceeded 10%. Immunoreactivity for Ki-67, synaptophysin, chromogranin A, and cytokeratin is well preserved in tissue with crush artifacts and can be interpreted reliably. The diagnosis of SCC should be questioned if fewer than 25% of cells show reactivity for Ki-67.

Topics: Artifacts; Biopsy; Bronchoscopy; Carcinoid Tumor; Carcinoma, Small Cell; Chromogranin A; Chromogranins; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Ki-67 Antigen; Lung Neoplasms; Synaptophysin

To study diagnostic significance of blood serum cancer antigens levels in patients who subsequently develop pulmonary cancer (PC) and gastrointestinal cancer (GIC).. ELISA was used to study expression of tumor antigens (CEA, NSE, CA19-9, CA242, AFP) in the blood serum of 27 PC and 31 GIC patients; 22 patients with lymphatic tumors and 32 patients with pulmonary and gastrointestinal inflammation served control. After removal of the tumor the same antigens including cytokeratines (CK) and differentiated leukocytic markers were studied immunocytochemically in the tumor cells with relevant monoclonal antibodies.. Sera of patients with verified afterwards cancer contained elevated concentrations of the antigens: NSE and CEA in PC, CA19-9, CA242 in GIC. The expression of these antigens including CK was found also in tumor cells of these patients. Atypical cells of lymphatic tumors had hemopoietic markers in the absence of CK. In inflammation and in lymphatic tumors, tumor antigens levels remained normal.. The test for tumor antigens levels in the serum may be used for early (preoperative) diagnosis of cancer, especially in tumors with difficult access or if they are asymptomatic.

Topics: Adenocarcinoma; Adult; Aged; Antibodies, Monoclonal; Antigens, Neoplasm; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Female; Gastrointestinal Neoplasms; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged

Detection of disseminated tumor cells in mediastinoscopic biopsies could improve staging and might be helpful concerning indications for neoadjuvant therapy regimens. This prospective study was performed to evaluate a simple and observer-independent polymerase chain reaction (PCR)-based method for the detection of disseminated tumor cells in regional lymph nodes.. Lymph nodes of 32 consecutive patients without neoadjuvant therapy were removed by systematic lymphadenectomy during resection of primary NSCLC. One hundred of these lymph nodes were cut into two equal halves which were examined using either routine histopathology or quantitative reverse transcriptase PCR (qRT-PCR). qRT-PCR amplification of cytokeratin 19 (CK19) transcripts was applied for the detection of tumor cell-specific RNA. We differentiated between illegitimate marker gene transcription and cancer-specific expression by using a cut-off value that was obtained from the analysis of 18 lymph nodes of patients with benign lung diseases. Subsequent to the evaluation of qRT-PCR, a pilot project with five additional patients was conducted to examine 19 mediastinoscopic biopsies, which were cut into two equal halves and proceeded as described above.. Ninety-four (94%) lymph nodes were tumor-free by histopathology. qRT-PCR detected disseminated tumor cells in 26 (28%) of these lymph nodes. All of the remaining six lymph nodes that were judged by the pathologist to contain tumor cells exhibited CK19 transcripts. Twenty-three patients had a pN0 status. qRT-PCR detected disseminated tumor cells in 13 (56%) of these pN0 patients. The mediastinoscopic biopsies showed disseminated tumor cells in four (21%) out of 19 histopathologically tumor-free samples.. CK19 qRT-PCR is a sensitive and specific tools for the detection of disseminated tumor cells in regional lymph nodes of patients with operable NSCLC. Further studies are required to asses if this molecular method might improve mediastinoscopic staging.

Topics: Aged; Biomarkers, Tumor; Biopsy; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratins; Lung Neoplasms; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Male; Mediastinoscopy; Mediastinum; Middle Aged; Neoplasm Staging; Prospective Studies; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm; Sensitivity and Specificity

Sarcomatoid carcinomas are rare malignancies which represent poorly differentiated epithelial tumors that may be difficult to recognize as such. While some cases may have obvious epithelial areas, the sarcomatoid areas are poorly distinguishable from true sarcoma at the light microscopic level and, by immunohistochemistry, often show only limited staining for traditional epithelial markers such as cytokeratin or epithelial membrane antigen. This can be particularly problematic for diagnosis on small biopsy specimens. We sought to assess the diagnostic utility of several immunohistochemical markers of epithelial differentiation including p63, MOC-31, and thyroid transcription factor-1 on sarcomatoid carcinomas of the head and neck (19 cases; 'spindle cell carcinomas'), lung (19 cases), and urinary bladder (11 cases). These results were compared to immunohistochemistry for the traditional epithelial markers pan-cytokeratin and epithelial membrane antigen. Staining for p63 showed the greatest diagnostic utility, positive in 63, 50, and 36% of head and neck, lung, and urinary bladder sarcomatoid carcinomas, respectively. p63 stains were positive in many cases where immunohistochemistry was negative for both pan-cytokeratin and epithelial membrane antigen. All three alternative markers were quite specific for epithelial differentiation, each staining less than 10% of the control group of 73 various primary and metastatic sarcomas, melanomas, and benign spindle cell lesions. In conclusion, immunostaining beyond traditional pan-cytokeratin and epithelial membrane antigen may have diagnostic utility in this context.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Melanoma; Membrane Glycoproteins; Membrane Proteins; Middle Aged; Mucin-1; Nuclear Proteins; Sarcoma; Sensitivity and Specificity; Thyroid Nuclear Factor 1; Transcription Factors; Urinary Bladder Neoplasms

Topics: Adult; Aged; Antigens, Neoplasm; Blood Proteins; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Proteomics; Reproducibility of Results; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

To evaluate five serum tumor markers used alone or in combination for the diagnosis of lung cancer.. The level of five serum tumor markers: NSE, pro-GRP, CYFRA21-1, p53 antibody and CEA was detected by ELISA in 50 healthy adults, 170 lung cancer patients and 60 patients with respiratory infection.. The level of the five serum tumor markers in lung cancer patients was significantly higher than that of healthy adults and patients with respiratory infection (P < 0.01). The level of NSE and pro-GRP in patients with small-cell lung cancer was significantly higher than those of the other subtypes of lung cancer (P < 0.01); The level of CYFRA21-1 in patients with squamous-cell carcinoma was significantly higher than that of other subtypes (P < 0.01). The specificity of p53 antibody was 100% in diagnosing lung cancer and the sensitivity of NSE, pro-GRP was much higher for small-cell lung cancer than for other subtypes (P < 0.01); The same was observed in CYFRA21-1 for the diagnosis of squamous-cell carcinoma (P < 0.01). The sensitivity of the tumor markers in diagnosing lung cancer was significantly enhanced if used in combination (P < 0.01).. These five tumor markers are valuable auxiliary parameters in diagnosing lung cancer. The combination of NSE and pro-GRP is more appropriate than other combinations in diagnosing small-cell lung cancer; the combination of CYFRA21-1, CEA and p53 antibody is the most valuable combination for diagnosing non-small-cell lung cancer. p53 antibody has the highest specificity for diagnosing lung cancer; CYFRA21-1 is the most valuable parameter for diagnosing squamous carcinoma.

Topics: Adenocarcinoma; Antibodies, Neoplasm; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Gastrin-Releasing Peptide; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Tumor Suppressor Protein p53

Currently, no satisfactory biomarkers are available to screen for lung cancer. Surface-Enhanced Laser Desorption/ionization Time-of-Flight Mass Spectrometry ProteinChip system (SELDI-TOF-MS) is one of the currently used techniques to identify biomarkers for cancers. The aim of this study is to explore the application of serum SELDI proteomic patterns to distinguish lung cancer patients from healthy individuals.. A total of 208 serum samples, including 158 lung cancer patients and 50 healthy individuals, were randomly divided into a training set (including 11 sera from patients with stages I/II lung cancer, 63 from patients with stages III/IV lung cancer and 20 from healthy controls) and a blinded test set (including 43 sera from patients with stages I/II lung cancer, 41 from patients with stages III/IV lung cancer and 30 from healthy controls). All samples were analyzed by SELDI technology. The spectra were generated on weak cation exchange (WCX2) chips, and protein peaks clustering and classification analyses were made using Ciphergen Biomarker Wizard and Biomarker Pattern software, respectively. We additionally determined Cyfra21-1 and NSE in the 208 serum samples included in this study using an electrochemiluminescent immunoassay.. Five protein peaks at 11493, 6429, 8245, 5335 and 2538 Da were automatically chosen as a biomarker pattern in the training set. When the SELDI marker pattern was tested with the blinded test set, it yielded a sensitivity of 86.9%, a specificity of 80.0% and a positive predictive value of 92.4%. The sensitivities provided by Cyfra21-1 and NSE used individually or in combination were significantly lower than that of the SELDI marker pattern (P < 0.005 or 0.05, respectively). Based on the results of the test set, we found that the SELDI marker pattern showed a sensitivity of 91.4% in the detection of non-small cell lung cancers (NSCLC), which was significantly higher than that in the detection of small cell lung cancers (P < 0.05); The pattern also had a sensitivity of 79.1% in the detection of lung cancers in stages I/II.. These results suggest that serum SELDI protein profiling can distinguish lung cancer patients, especially NSCLC patients, from normal subjects with relatively high sensitivity and specificity, and the SELDI-TOF-MS is a potential tool for the screening of lung cancer.

Topics: Adult; Aged; Algorithms; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cations; Decision Trees; Female; Gene Expression Regulation, Neoplastic; Humans; Immunoassay; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Protein Array Analysis; Proteomics; Sensitivity and Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Carcinoma, Endometrioid; Female; Humans; Immunohistochemistry; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

To set up a molecular method (i.e. RT-PCR) that can be used to detect disseminated tumor cells (DTCs) in regional lymph nodes (LNs) in patients with lung cancer and to evaluate its clinical significance.. Cytokeratin 19 (CK(19)) was used as marker. Serial dilution study for LC-5 cells (a lung squamous cell line) was performed to detect sensitivity of the molecular protocol. Regional LNs (n = 261) and primary lung cancer tissue (n = 40) were obtained from 40 patients with lung cancer who underwent lobectomy or pneumonectomy. They were randomly categorized into two groups: group I (LN-based study, n = 20) and group II (patient-based study, n = 20). Each LN was halved. One half of a LN was subjected to histological examination (HE) and the other half was subjected to RT-PCR amplification of CK(19) mRNA. The effect on survival was analyzed. The cumulative survival was calculated by the Kaplan-Meier method and compared by the log rank test. The Cox model analyzed the prognostic factors.. CK(19) mRNA expressed in all tumor tissues as well as LC-5, PAa cells (a lung adenocarcinoma cell line), but not in normal control LNs. Serial dilution study for LC-5 cells demonstrated that CK(19) mRNA was detectable at a concentration as low as 10 LC-5 cells in 1x10(7) LN cells. There was no significant difference between the detecting result of single LN and that of mixed LNs (P > 0.05). In 18 of 40 patients, the metastasis in regional LNs was found by both HE and RT-PCR. Of 22 patients without pathologically involved nodes, six (27%) were found to express CK(19) mRNA in regional LNs. According to the results of regional LNs in 40 patients by molecular assay, the presence of the CK(19) product in LNs was related to tumor size (chi(2) = 5.76, P < 0.025) as well as cell differentiation of the tumor (chi(2) = 7.08, P < 0.01). Following a median observation time of 26 months (range, 4 to 60 months), patients with DTCs in nodes showed significant shorter disease-free survival duration than node-negative patients (log-rank test, P = 0.001). The independence of this prognostic significance was demonstrated by a multivariate analysis (Cox regression model, P = 0.004). The results diagnosed by HE had no significant effect on prognoses (P = 0.455).. Comparing with HE, RT-PCR can make more accurate assessment of metastatic status in LNs, which is helpful for screening the patients in whom the early subclinical metastasis exists and disclosing the intrinsic regulation of malignant metastasis. The presence of DTCs in LNs is an independent factor for prognosis. Molecular detection of DTCs in LNs is a supplement for current tumor staging in lung carcinoma.

Topics: Aged; Biomarkers, Tumor; Cell Line, Tumor; Female; Humans; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Survival Analysis

Overwhelming evidence has demonstrated tobacco smoke (TS) is causally associated with various types of cancers, especially lung cancer. Sustained epithelial cell hyperplasia and squamous metaplasia are considered as preneoplastic lesions during the formation of lung cancer. The cellular and molecular mechanisms leading to lung cancer due to TS are not clear. Mitogen-activated protein kinases (MAPK)/activator protein-1 (AP-1) can be activated by various stimuli and play a critical role in the control of cell proliferation and differentiation. To date, information on the response of the MAPK/AP-1 pathway during hyperplasia and squamous metaplasia induced by TS is lacking. We therefore investigated the effects of TS on the development of epithelial hyperplasia and squamous metaplasia, regulation of MAPK/AP-1 activation, and expression of AP-1-regulated cell cycle proteins and differentiation markers in the lungs of rats. Exposure of rats to TS (30 mg/m(3) or 80 mg/m(3), 6 h/day, 3 days/week for 14 weeks) dramatically induced cell proliferation and squamous metaplasia in a dose-dependent manner, effects that paralleled the activation of AP-1-DNA binding activity. Phosphorylated ERK1/2, JNK, p38 and ERK5 were significantly increased by exposure to TS, indicating the activation of these MAPK pathways. Expression of Jun and Fos proteins were differentially regulated by TS. TS upregulated the expression of AP-1-dependent cell cycle proteins including cyclin D1 and proliferating cell nuclear antigen (PCNA). Among the AP-1-dependent cell differentiation markers, keratin 5 and 14 were upregulated, while loricrin, filaggrin and involucrin were downregulated following TS exposure. These findings suggest the important role of MAPK/AP-1 pathway in TS-induced pathogenesis, thus providing new insights into the molecular mechanisms of TS-associated lung diseases including lung cancers.

Topics: Animals; Cell Proliferation; Cyclin D1; Enzyme Activation; Filaggrin Proteins; Hyperplasia; Intermediate Filament Proteins; JNK Mitogen-Activated Protein Kinases; Keratins; Lung Neoplasms; Male; Membrane Proteins; Metaplasia; Mitogen-Activated Protein Kinases; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Proliferating Cell Nuclear Antigen; Protein Precursors; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-jun; Rats; Rats, Inbred WKY; Signal Transduction; Smoking; Transcription Factor AP-1

To evaluate the clinical and prognostic significance of the tumor markers cytokeratin 19 fragment, carcinoembryonic antigen and neuron-specific enolase in lung cancer patients.. Serum levels of cytokeratin 19 fragment, carcinoembryonic antigen and neuron-specific enolase were measured using electrochemical luminescence immunoassay in 46 lung cancer patients. Serum levels of cytokeratin 19 fragment, carcinoembryonic antigen, and neuron-specific enolase higher than 3.6 ng/ml, 5.0 ng/ml and 13.0 ng/ml, respectively, were considered as elevated.. Cytokeratin 19 fragment, carcinoembryonic antigen, and neuron-specific enolase were elevated in 19.6%, 43.5%, and 63% of patients, respectively. Elevated levels of neuron-specific enolase were detected more frequently in smokers than in ex-smokers (p=0.003). Likewise preoperative levels of carcinoembryonic antigen (p=0.023) and neuron-specific enolase (p=0.007) were statistically higher in smokers than in ex-smokers. A significant correlation was detected between the level of cytokeratin 19 fragments and smoking cumulative exposure (r=0.542, p=0.037). The number of patients with elevated levels of cytokeratin 19 fragment and neuron-specific enolase was higher in more advanced disease than in early lung cancer (p=0.036 and p=0.036, respectively). Preoperative levels of cytokeratin 19 fragment (p=0.017 and p=0.016, respectively) and neuron-specific enolase (p=0.03 and p=0.006, respectively) were significantly associated with more advanced disease and tumor size, as well as tumor histology in non-small cell lung cancer (p=0.03 and p=0.016, respectively). Preoperative levels of cytokeratin 19 fragments were higher in squamous cell carcinoma than in adenocarcinoma (p=0.026). Elevated preoperative serum levels of cytokeratin 19 fragment predict a poor prognosis for lung cancer patients (p=0.007).. Alteration of serum tumor markers cytokeratin 19 fragment, carcinoembryonic antigen and neuron-specific enolase is associated with particular tumor histology, smoking habit, more advanced disease and poor prognosis.

Topics: Adenocarcinoma; Adult; Aged; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Phosphopyruvate Hydratase; Prognosis; Smoking

We sought to determine whether cytokeratin-positive cells can be detected as markers of lymph node metastasis by using flow cytometry within a time frame suitable for intraoperative decision making in non-small cell lung cancer.. Five lymph nodes from each of 20 patients with non-small cell lung cancer were randomly selected for study. Each node was divided longitudinally into 3 pieces: one piece for flow cytometry, one for immunohistochemical staining, and the last for conventional hematoxylin and eosin staining. In both flow cytometry and immunohistochemistry, cytokeratin-positive cells were detected with the fluorescein isothiocyanate-conjugated anti-cytokeratin antibody AE1/AE3.. Cytokeratin-positive nodes were detected by means of flow cytometry within 40 minutes. Eight (8%) of the 100 lymph nodes from 4 (20%) of the 20 patients were deemed positive for metastasis on the basis of conventional histologic examination. By contrast, 33 (33%) lymph nodes from 13 (65%) patients were deemed positive on the basis of immunohistochemical cytokeratin staining, and 38 (38%) lymph nodes from 14 (70%) patients were deemed positive on the basis of flow cytometric cytokeratin-positive cell detection. All nodes deemed positive for metastasis on the basis of conventional and immunohistochemical methods were also positive on flow cytometry.. Flow cytometry enables rapid intraoperative diagnosis of nodal metastasis in patients with non-small cell lung cancer. Flow cytometric detection of cytokeratin-positive cells within lymph nodes correlates with their immunohistochemical detection, and its level of sensitivity is greater than that of conventional histologic staining and about equal to that of immunohistochemical staining.

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Flow Cytometry; Humans; Immunohistochemistry; Intraoperative Period; Keratins; Lung Neoplasms; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis

Small cell carcinoma is a rare malignant disease with high mortality, which is pathologically diagnosed by using routine neuroendocrinal markers, such as neuron-specific enolase (NSE), synaptophysin (SYN), chromogranin A (CgA). This study was to investigate the expression of CD56, a neural cell adhesion molecule (NCAM), in small cell carcinoma tissues, and to explore the possibility of CD56 as a diagnostic marker of small cell carcinoma.. Eighty samples of small cell carcinoma were collected, including 42 samples of small cell lung carcinoma (with 20 cases of lymph node metastases), 21 samples of small cell esophageal carcinoma, and 17 samples of small cell colorectal carcinoma. Thirty-eight samples of non-small cell lung cancer (with 28 cases of lymph node metastases), including 26 samples of squamous cell carcinoma and 12 samples of adenocarcinoma, were used as control. All samples were detected using markers of CD56, NSE, SYN, CgA, cytokeratin (CK), and epithelial membrane antigen (EMA) immunohistochemically.. Positive rate of CD56 was significantly higher in either small cell lung carcinoma or their metastatic lymph nodes than in either non-small cell lung carcinoma or their metastatic lymph nodes [90.5% (38/42) vs. 7.8% (3/38), 90.0% (18/20) vs. 3.5% (1/28); H=85.731, P<0.001]. Positive rate of CD56 in small cell carcinoma samples (86.3%, 69/80) were significantly higher than those of SYN (78.8%, 63/80), CgA (73.8%, 59/80), EMA (66.3%, 53/80), CK (61.3%, 49/80), and NSE (56.3%, 45/80) (H=38.871, P<0.001). Positive rate of CD56 in small cell lung carcinoma (90.5%, 38/42) was similar to those in small cell esophageal carcinoma (81.0%, 17/21) and small cell colorectal carcinoma (82.4%, 14/17) (H=1.651, P=0.438).. CD56 is highly expressed in either small cell carcinoma or their metastatic lymph nodes without organ-specificity. It could serve as a potential diagnostic marker of small cell carcinoma.

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; CD56 Antigen; Chromogranin A; Colorectal Neoplasms; Diagnosis, Differential; Esophageal Neoplasms; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Lymphatic Metastasis; Mucin-1; Phosphopyruvate Hydratase; Synaptophysin

To assess the diagnostic values of carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and cytokeratin 19 fragments (CYFRA 21-1) as markers of pleurisy in primary lung cancer.. Prospective case-control study.. A tertiary university hospital.. Thirty-four patients with lung cancer and 16 patients with tuberculous pleurisy.. Levels of CEA, NSE, and CYFRA 21-1 were measured by immunoassay in the serum and pleural fluid of patients with lung cancer and of patients with tuberculous pleurisy. Patients with lung cancer were found to have significantly higher serum and pleural fluid levels of CEA and CYFRA 21-1 than patients with tuberculous pleurisy. Using cutoff values of 5 ng/mL, 20 ng/mL, and 3.3 ng/mL for serum CEA, NSE, and CYFRA 21-1, respectively, the sensitivities and specificities of these tumor markers were as follows for differentiating malignant effusion from benign: CEA, 68% and 93%; NSE, 34% and 93%; and CYFRA 21-1, 45% and 100%. Using cutoff values of 5 ng/mL, 20 ng/mL, and 45 ng/mL for pleural fluid, the sensitivities and specificities were as follows: CEA, 82% and 94%; NSE, 36% and 94%; and CYFRA 21-1, 61% and 81%. A combination of pleural fluid CEA and NSE increased sensitivity and specificity.. In the diagnosis of malignant effusion associated with lung cancer, the determinations of CEA and NSE in pleural fluid could enhance diagnostic yield better than those of all three tumor markers.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Case-Control Studies; Humans; Keratin-19; Keratins; Lung Neoplasms; Peptide Fragments; Phosphopyruvate Hydratase; Pleural Effusion; Sensitivity and Specificity

The human epidermal growth factor receptor (EGFR) is reportedly overexpressed in 15-20% of breast carcinomas. EGFR overexpression is associated with reduced survival and is inversely correlated with expression of estrogen receptor (ER). This study assessed EGFR expression in breast carcinomas with squamous differentiation. The immunohistochemical (IHC) expression of EGFR was evaluated in 39 breast carcinomas with squamous differentiation (30 pure squamous, 6 adenosquamous, 3 carcinosarcomas) by use of the pharmDx assay (clone 2-18C9, DakoCytomation). Cases were considered positive if at least 10% of the cells showed 1+ positivity in the squamous component. Squamous differentiation was confirmed with IHC for CK5-6 (clone D5/16B4, DakoCytomation). ER, PR, and HER2 status as well as clinical information regarding treatment and outcome were correlated. As a control, a tissue microarray comprising 280 lymph node positive breast carcinomas was evaluated with the same EGFR assay. The 39 patients ranged in age from 33 to 77 years (mean 52). The tumors measured 1.3-30 cm (mean 4.8). Sentinel or full axillary lymph node dissection was performed in 28 patients. Fourteen patients had positive lymph nodes. At the time of initial diagnosis, 3 patients had distant metastasis. Follow-up was available for 16 patients (mean 45 months). Disease-free survival at 3 years was 70%. Among the 39 tumors 87% (34) were positive for EGFR (p<0.0001). Sixty-nine percent (27 of 39) showed >50% 2+ EGFR staining. EGFR-positive tumor cells (showing squamous morphology) were also found in 1 bone, 1 lung, and 8 of 11 lymph node metastases available for evaluation. All 11 lymph nodes showed squamous differentiation. All but 1 of the EGFR+ tumors examined were ER and PR negative. Six EGFR-positive tumors were HER2 positive. No statistically significant differences in HER2 status, size, lymph node status and disease-free survival were observed between EGFR+ and EGFR- cases, but the number of EGFR-negative tumors was quite small. Nine of 280 (3%) of lymph node-positive invasive carcinomas on the tissue microarray were EGFR+; review of the initial diagnostic slides failed to reveal squamous features in all but 1 of the 9 EGFR+ tumors. Breast carcinomas with squamous differentiation are a distinct subgroup of breast tumors with a very high frequency of EGFR positivity. Breast carcinomas of this type would be ideal candidates for a trial with EGFR inhibitors.

Topics: Adult; Aged; Bone Neoplasms; Breast Neoplasms; Carcinoma, Adenosquamous; Carcinoma, Squamous Cell; Carcinosarcoma; Cell Differentiation; ErbB Receptors; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Middle Aged; Neoplasm Staging; Receptor, ErbB-2; Receptors, Estrogen

The aim of the study was to assess the role of serum tumour markers (NSE, Cyfra 21-1, CEA, LDH, ferritin) as a prognostic and predictive factors in 79 patients with advanced NSCLC treated with chemotherapy. Objective response to treatment was significantly more frequent in the patient with serum NSE > 12.5 ng/ml. Progression of disease was observed more often in patients with serum Cyfra 21-1 >10 ng/ml or LDH >480 U/L. CEA >3 ng/ml, LDH >480 U/L, for coefficient >1, NSE >20 ng/ml and Cyfra 21-1 >10 ng/ml had a negative impact on survival in univariate analysis. Independent negative prognostic significance of fer coefficient >1 was confirmed by multivariate analysis.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Mucin-1; Neoplasm Proteins; Predictive Value of Tests; Prognosis; Serum; Survival Analysis; Survival Rate

To better understand the mechanism underlying the hepatocellular carcinoma (HCC) metastasis and to search potential markers for HCC prognosis, differential proteomic analysis on two well-established HCC cell strains with high and low metastatic potentials, MHCC97-H and MHCC97-L, was conducted using two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/time-of-flight mass spectrometry. Cytokeratin 19 (CK19) was identified and found to be overexpressed in MHCC97-H as compared with MHCC97-L. This result was further confirmed by two-dimensional Western blot analysis and immunofluorescence assay. Furthermore, one-dimensional Western blot analysis showed consistently increased CK19 expression in progressively more metastatic cells. Immunohistochemical study on 102 human HCC specimens revealed that more patients in the CK19-positive group had overt intrahepatic metastases (satellite nodules, p < 0.05; vascular tumor emboli, p < 0.001; tumor node metastatis staging, p < 0.001). CK19 fragment CYFRA 21-1 levels measured in sera from nude mice model of human HCC metastasis with radioimmunoassay increased in parallel with tumor progression and rose remarkably when pulmonary metastases occurred. The results demonstrated that overexpression of CK19 in HCC cells is related to metastatic behavior. Serum CK19 level might reflect the pathological progression in some HCC and may be a useful marker for predicting tumor metastasis and a therapeutic target for the treatment of HCC patients with metastases.

Topics: Animals; Biomarkers, Tumor; Carcinoma, Hepatocellular; Cell Line, Tumor; Electrophoresis, Gel, Two-Dimensional; Humans; Keratins; Liver Neoplasms; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Metastasis; Peptide Fragments; Proteome; Random Allocation; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The correlation between detection of occult neo-plastic cells (ONCs) in lymph node sinuses and the recurrence/metastasis of primary breast, lung, esophageal, and gastric cancer was examined. Among patients with Stage I-III cancer treated by radical resection with dissection of at least 10 lymph nodes, 40 patients who suffered recurrence/metastasis within 5 years post-operatively and 94 patients who did not have recurrence within 5 years were randomly selected. A total of 1,340 lymph nodes were subjected to immunohistochemical staining for cytokeratin to identify ONCs. Then the sensitivity, specificity, positive predictive value, and negative predictive value of ONCs were determined for predicting the recurrence of each cancer. These parameters were respectively 40.0, 75.9, 62.4, and 55.8% for breast cancer, while the respective values were 50.0, 77.4, 68.9, and 60.8% for lung, 57.1, 64.3, 61.5, and 60.0% for esophageal, and 68.8, 65.0, 66.3, and 67.5% for gastric cancer. All of the parameters exceeded 65% for gastric cancer. ONCs also showed a high specificity for breast and lung cancer, but both the sensitivity and specificity were low for esophageal cancer.

Topics: Breast Neoplasms; Esophageal Neoplasms; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Neoplasm Recurrence, Local; Neoplasms; Predictive Value of Tests; Random Allocation; Sensitivity and Specificity; Single-Blind Method; Stomach Neoplasms

The aim of the present study was to examine the impact of sputum carcinoembryonic antigen (CEA), neuron-specific enolase (NSE) and cytokeratin fragment 19 (CYFRA 21-1) levels in lung cancer diagnosis and to compare the diagnostic usefulness of sputum assays with that of serum assays.. Forty-seven patients with lung cancer and 62 with benign lung disease were studied. Tumour marker levels in sputum (sp.) and serum (ser) were measured by immunoradiometric assays.. Sputum and serum tumour marker levels were significantly higher in lung cancer than in benign disease. When the specificity was 95%, the sensitivity was 57%, 43%, 36%, 30%, 28% and 19%, for spCEA, serCYFRA 21-1, spCYFRA 21-1, serCEA, serNSE, and spNSE, respectively. Bayesian analysis showed that the best predictive values correspond to spCEA and serCYFRA 21-1. The maximum overall gain was obtained in pretest probability of 0.35 for both spCEA and serCYFRA 21-1, with predictive values of 84% and 80% for spCEA and serCYFRA 21-1, respectively.. Sputum tumour marker levels were no more useful than the serum levels in lung cancer diagnosis. SpCEA offered the best predictive values but these were still not sufficiently satisfactory for spCEA to be proposed for routine use.

Topics: Adult; Aged; Aged, 80 and over; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Cross-Sectional Studies; Female; Humans; Immunoradiometric Assay; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; ROC Curve; Sensitivity and Specificity; Specimen Handling; Sputum

We investigated 27 pleomorphic carcinomas of the lung for exon 1 K-ras gene mutations using polymerase chain reaction-single-strand conformation polymophism analysis and direct sequencing. All pleomorphic carcinomas were biphasic, that is, composed of an adeno-, squamous- or large-cell-carcinomatous component associated with a spindle- and/or giant-cell component. Of 27 cases, six (22%) showed K-ras codon 12 mutations, which is a figure higher than that previously reported on in pure sarcoma-like pleomorphic carcinomas. Five tumors displayed the same mutation in both the epithelial and the sarcomatoid components, whereas in one tumor the mutation was restricted to the epithelial component. All mutations occurred in smokers, and were transversions, including GGT (glycine) to TGT (cysteine) change in two cases, to GCT (alanine) in two and to GTT (valine) in two. No significant relationships were found between the occurrence and type of mutations and patients' survival or any other clinicopathological variable, suggesting that K-ras mutations are early events in the development of these tumors. Our results indicate that most, though not all, biphasic pleomorphic carcinomas of the lung are monoclonal in origin, and that cigarette smoking may have a causative role in the development of K-ras alterations in these tumors, as all mutations are transversions.

Topics: Adenocarcinoma; Adult; Aged; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Clone Cells; DNA Mutational Analysis; DNA, Neoplasm; Female; Genes, ras; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Mutation; Mutation, Missense; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Smoking; Survival Analysis; Vimentin

An 11-year-old male Collie was presented with a swelling of the face caused by tumor masses arising from the gingiva. Postmortem examination revealed metastases to the lymph nodes, lung, liver, and orbital cavity. Histologically, the tumor represented a combination of fibrosarcomatous proliferation, pulpal mesenchyme, and undifferentiated odontogenic epithelium, with a follicular or plexiform growth pattern. In addition, the follicular areas of the tumor showed a biphasic character, and there were numerous apoptotic cells in plexiform areas. Furthermore, acidophilic material resembling dysplastic dentine or enamel matrix was observed in the metastatic lesion in the lung. Based on the histological characters, the present case was diagnosed as malignant ameloblastic fibro-odontoma. This study is the first known description of a possible malignant ameloblastic fibro-odontoma in a dog with metastasis to distant organs.

Topics: Animals; Chromogenic Compounds; Dog Diseases; Dogs; Gingival Neoplasms; Immunohistochemistry; In Situ Nick-End Labeling; Keratins; Lung Neoplasms; Odontoma

A case history is presented of a 53-year-old woman with an incidental finding of a breast lump, identified after having had chemotherapy for lung metastases from a rectal carcinoma. Clinical examination, ultrasound, mammography, fine needle aspiration and core biopsies could not prove definitively whether the breast lump represented a metastasis from colorectal carcinoma. Following local excision, the final diagnosis of metastatic colorectal carcinoma to the breast was based on the absence of any site of origin within the breast (i.e. no surrounding DCIS) and on the expression of cytokeratin CK7 and CK20 on immunohistochemistry. Postoperative chemotherapy was initiated. Four months later, although without local recurrence in the breast, the patient developed cutaneous metastatic deposits and active treatment was stopped. A review of other cases of breast metastases from extramammary sources is presented. Possible mechanisms for this rare and unusual phenomenon are discussed.

Topics: Adenocarcinoma; Biomarkers; Breast Neoplasms; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Middle Aged; Skin Neoplasms

We studied the staining patterns of bronchioloalveolar carcinoma (BAC) with antibodies to cytokeratin (CK) 7, CK20, and thyroid transcription factor-1 (TTF-1) to determine the diagnostic usefulness of this panel in differentiating BAC from metastatic adenocarcinoma in material obtained by fine-needle aspiration biopsy (FNAB) of the lung. We identified 16 cases of BAC. Of these, 6 were mucinous, 4 were nonmucinous, and 6 were mixed with focal mucinous differentiation. Immunohistochemical analysis with antibodies to CK7, CK20, and TTF-1 was performed on cell-block sections. Of the 6 mucinous BACs, 4 (67%) were CK7+, CK20+, and TTF-1-. All 4 nonmucinous BACs were CK7+ and CK20-, and 2 (50%) were TTF-1+. All 6 mixed BACs were diffusely positive for CK7 and focally positive for CK20; 5 (83%) were TTF-1+. Nonmucinous BACs display CK7, CK20, and TTF-1 immunoreactivity similar to conventional pulmonary adenocarcinoma. Mucinous and mixed BACs have an immunohistochemical phenotype that is different from that of conventional pulmonary adenocarcinoma. Knowledge of these staining patterns is crucial for distinguishing mucinous and mixed BACs from metastatic adenocarcinoma involving the lungs.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy, Fine-Needle; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

In 1999, the World Health Organization redefined bronchioloalveolar carcinomas (BACs) as those neoplasms with only a pure lepidic growth pattern and no invasion.. The present study examined 45 lung cancers with a BAC component (1) to determine whether these tumors would be classified as BACs by current World Health Organization standards, (2) to quantitate the BAC component within these tumors, and (3) to see if phenotypic differences exist between the so-called invasive and noninvasive regions of these tumors.. Retrospective review of hematoxylin-eosin-stained slides and classification of histologic grade, tumor subtype, and percentage of pure BAC pattern, with further characterization by immunohistochemical staining for thyroid transcription factor 1, cytokeratin 7, cytokeratin 20, and Ki-67 antibodies.. Only 7 (15.6%) of the 45 tumors examined could be classified as BAC by current strict World Health Organization criteria. Those tumors, classified as nonmucinous and mixed, showed similar immunohistochemical staining for cytokeratin 7, cytokeratin 20, and thyroid transcription factor 1; mucinous tumors showed disparate staining. Significant differences in immunohistochemical staining and tumor cell proliferation were seen for the regions of tumors designated as lepidic, infiltrative, and leading edge and for the regions of tumors with different histologic grades (ie, well, moderately, and poorly differentiated).. Nonmucinous and mixed BACs are phenotypically similar and show identical immunohistochemical staining patterns; mucinous tumors, on the other hand, show disparate immunohistochemical staining. Pulmonary neoplasms designated as adenocarcinomas with a BAC component represent a heterogenous group with a range of cell types, differentiation, growth, and immunophenotypes. Within an individual neoplasm, there are regional differences in these parameters as well.

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Adenocarcinoma, Mucinous; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cell Differentiation; Connective Tissue; Female; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Proteins; Nuclear Proteins; Phenotype; Retrospective Studies; Staining and Labeling; Thyroid Nuclear Factor 1; Transcription Factors

We examined the effect of stable transfection of dominant negative TbetaR-II (dn TbetaR-II) cDNA in a human oral carcinoma cell line that contained normal Ras and was growth inhibited by TGF-beta1. Two clonal cell lines containing dn TbetaR-II were isolated and compared to the vector-only control and parent cell line. The treatment of cells with exogenous TGF-beta1 resulted in a decrease in ligand-induced growth inhibition and loss of c-myc downregulation in test cells compared to controls; transcriptional activation of certain genes including fra-1 and collagenase was retained. Cells containing dn TbetaR-II grew faster in monolayer culture, expressed less keratin 10 and exhibited increased motility and invasion in vitro compared to control cell lines. Endogenous TGF-beta1 production and the regulation of MMP-2 and MMP-9 by TGF-beta1 remained unchanged. After orthotopic transplantation to the floor of the mouth in athymic mice, cells containing dn TbetaR-II formed comparable numbers of primary tumours at the site of inoculation as controls but the tumours were less differentiated as demonstrated by the absence of keratin 10 immunostaining. Further, metastatic dissemination to the lungs and lymphatics was more evident in grafts of cells containing dn TbetaR-II than controls. Taken together, the results demonstrate that attenuation of TGF-beta signalling through transfection of dn TbetaR-II cDNA leads to an enhanced growth rate, a loss of tumour cell differentiation and an increase in migration and invasion, characteristics that corresponded to the development of the metastatic phenotype.

Topics: Cell Differentiation; Cell Division; Cell Line, Tumor; Cell Movement; Humans; Keratin-10; Keratinocytes; Keratins; Lung Neoplasms; Mouth Neoplasms; Neoplasm Metastasis; Phenotype; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Signal Transduction; Transforming Growth Factor alpha

Cytokeratin 19 fragment (CYFRA 21-1) level in serum have already been documented as a useful tumor marker for lung cancer. In the present study, we hypothesized that CYFRA 21-1 increases in the sera of patients with radiation pneumonitis, resulting from epithelial cell damage. We measured CYFRA 21-1 in the sera of patients with radiation pneumonitis and evaluated the correlation between CYFRA 21-1 level and severity of radiation pneumonitis as well as clinical course. We studied 16 patients diagnosed with radiation pneumonitis associated with primary lung cancer. CYFRA 21-1 levels in the sera of patients with diffuse radiation pneumonitis (n = 6) significantly increased compared to normal smokers (n = 10) or patients with local radiation pneumonitis (n = 10). CYFRA 21-1 values in sera changed according to the progression or improvement of the diffuse radiation pneumonitis. An immunohistochemical study using pulmonary tissues obtained from autopsied patients with radiation pneumonitis demonstrated that the hyaline membrane and proliferating type II pneumocytes were strongly stained by the anti-human cytokeratin 19 antibody. Our data demonstrated that CYFRA 21-1 was increased in patients with diffuse radiation pneumonitis. Since CYFRA 21-1 is widely used as a tumor marker for lung cancer, this evidence should be noted especially in irradiated patients.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers; Epithelial Cells; Female; Humans; Immunohistochemistry; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Radiation Pneumonitis; Treatment Outcome

To assess the value of Cyfra 21-1, carcino-embryonic antigen (CEA) and neuron-specific enolase (NSE) combined, all three together as prognostic factors in advanced stage non-small cell lung cancer (NSCLC) patients.. Serum samples from untreated NSCLC patients were prospectively collected. All assays were performed using commercial kits blind to clinical information. Serum levels of CEA, NSE and Cyfra 21-1 higher than 10, 13 and 3.5 ng/ml, respectively, were considered as elevated.. 264 patients (men, 87%), with Performans Status (PS) of 0/1 in 80% and stage IV disease in 65% were studied. Cyfra 21-1, CEA and NSE were elevated in 52.5%, 41.8% and 33.2% of patients, respectively. Median survival was 9 months (range, 1-77). Cyfra 21-1, age, PS, stage as well as the combination of the three markers together correlated with prognosis in univariate analysis. Multivariate analysis demonstrated that age > or = 65 years (HR = 1.3 [1.02-1.70], p = 0.03), PS 2 (HR = 4.3 [3.13-6.11], p < 0.0001), Cyfra 21-1 > or = 3.5 ng/ml (HR = 1.3 [1.06-1.78], p = 0.01) and the combination of the three markers (HR = 1.06 [1.009-1.13], p = 0.02) remained prognostic determinants.. Combining Cyfra 21-1, NSE and CEA correlated with prognosis in a significant and independent manner.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Prognosis; Prospective Studies; Survival Analysis

The purpose of this study was to assess the feasibility of using rare event imaging system (REIS)-assisted analysis to detect occult tumor cells (OTCs) in peripheral blood (PB). The study also sought to determine whether REIS-assisted OTC detection presents a clinically viable alternative to manual microscopic detection to establish the true significance of OTC from solid epithelial tumors.. We recently demonstrated proof of concept using a fluorescence-based automated microscope system, REIS, for OTC detection from the PB. For this study, the prototype of the system was adopted for high-throughput and high-content cellular analysis.. The performance of the improved REIS was examined using normal blood (n = 10), normal blood added to cancer cells (n = 20), and blood samples obtained from cancer patients (n = 80). Data from the screening of 80 clinical slides from breast and lung cancer patients, by manual microscopy and by the REIS, revealed that as many as 14 of 35 positive slides (40%) were missed by manual screening but positively identified by REIS. In addition, REIS-assisted scanning reliably and reproducibly quantified the total number of cells analyzed in the assay and categorized positive cells based on their marker expression profile.. REIS-assisted analysis provides excellent sensitivity and reproducibility for OTC detection. This approach may enable an improved method for screening of PB samples and for obtaining novel information about disease staging and about risk evaluation in cancer patients.

Topics: Breast Neoplasms; Carcinoma, Small Cell; Cell Count; Cell Line, Tumor; Humans; Image Processing, Computer-Assisted; Immunohistochemistry; Keratins; Lung Neoplasms; Microscopy, Fluorescence; Neoplastic Cells, Circulating; Reproducibility of Results; Sensitivity and Specificity

To assess the diagnostic accuracy and to study the histologic typing of mediastinal lesions using core needle biopsies.. The histopathology and immunophenotype of 65 mediastinal core needle biopsy specimens were studied retrospectively by light microscopy and immunohistochemical staining (ABC method). Gene rearrangement studies were performed in some of the non-Hodgkin's lymphomas cases using PCR. Follow-up records were also analyzed.. Morphologically, all specimens showed a combination of epithelioid cells, lymphoid cells and fibrous tissue in different proportions. The pathologic diagnoses included lymphoma (21 cases), pulmonary carcinoma (20 cases), thymoma (14 cases), thymic carcinoma (4 cases), seminoma (3 cases) and chronic inflammation (1 case). Definitive diagnosis was not possible in 2 cases due to insufficient material. The tumor cells in lymphoma (21 cases) expressed CD20, CD3, TDT, CD30, CD15 or EMA, depending on their histologic subtypes. Tumor cells in the 17 pulmonary carcinoma cases expressed cytokeratin (CK), except 3 cases of small cell carcinoma of lung. Synaptophysin, chromogranin A and neuron-specific enolase were all positive in the 10 cases of small cell carcinoma of lung and 1 case of thymic small cell carcinoma (which was also CD5 negative). The 3 cases of adenocarcinoma of lung showed positivity for thyroid transcription factor-1 (TTF-1) and they were negative for CD5. The 14 thymoma cases expressed CK, CD3 or CD20. The 3 thymic carcinoma cases expressed CK and CD5. Placental-like alkaline phosphatase (PLAP) was positive in 3 seminoma cases which were CK-negative. Immunoglobulin heavy chain gene was rearranged in the 3 cases of diffuse large B-cell lymphoma and 1 B-cell anaplastic large cell lymphoma case. T-cell receptor beta gene was rearranged in 5 T-cell lymphoblastic lymphoma cases.. Microscopic assessment of tissue samples from mediastinal core needle biopsies should be made in combination with clinical and radiologic information. Ancillary investigations, including immunohistochemical staining and/or gene rearrangement studie, are needed in both non-lymphoma and lymphoma cases of mediastinum.

Topics: Adolescent; Adult; Aged; Biopsy, Needle; CD5 Antigens; Child; Child, Preschool; Diagnosis, Differential; Female; Follow-Up Studies; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; Humans; Keratins; Lung Neoplasms; Lymphoma; Male; Mediastinal Diseases; Mediastinum; Middle Aged; Retrospective Studies; Thymus Neoplasms

Telomerase is a ribonucleoprotein complex mainly composed of a reverse transcriptase catalytic subunit (telomerase reverse transcriptase gene, hTERT) that copies a template region of its RNA subunit to the end of the telomere. For detecting telomerase activity in a tissue specimen the TRAP assay is a relatively sensitive and specific method, but it can be used only on fresh tissue extracts and offers no information at the single cell level. Immunohistochemistry (IHC) allows to detect hTERT protein expression at an individual cell level in human tissues. We have tested commercially available anti-hTERT antibodies in formalin-fixed and paraffin-embedded human tissues by IHC. Only one monoclonal antibody (NCL-hTERT; Novacastra) was sufficiently specific and this was applied to human tissues in which telomerase activity had been shown by TRAP assay and hTERT mRNA expression by RT-PCR. hTERT protein localized diffusely in the nucleoplasm and more intensely in the nucleoli of cancer cells and proliferating normal cells. Mitotic cells showed diffuse staining of the entire cell. Granular cytoplasmic staining was occasionally found in some tumor cells. In telomerase-positive tumors not all the tumor cells showed hTERT immunoreactivity. A significantly heterogeneous hTERT protein expression was observed in human tumor tissues. The hTERT immunostaining in fixed tissues was concordant with telomerase activity and hTERT mRNA expression in corresponding non-fixed samples. Quantitative RT-PCR of microdissected sections showed that hTERT mRNA expression was higher in cells with nuclear expression than in those with cytoplasmic expression. Double staining with the M30 antibody showed that a subpopulation of hTERT-negative cells is apoptotic. We conclude that: (1) hTERT protein can be detected by IHC in fixed human tissues, but the choice of the antibody, tissue processing, and reaction conditions are critical, (2) hTERT protein localizes in the nucleoplasm, more strongly in the nucleolus, and occasionally in the cytoplasm, (3) telomerase-positive tumors show significant heterogeneity of hTERT protein expression, and (4) a subpopulation of hTERT protein negative tumor cells is identified as apoptotic cells.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Apoptosis; Breast Neoplasms; Cell Nucleolus; Cell Nucleus; Colonic Neoplasms; Cytoplasm; DNA-Binding Proteins; Female; Gene Expression; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphocytes; Male; Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm; Sarcoma; Spermatogonia; Telomerase; Urinary Bladder Neoplasms

We describe a case of primary pulmonary paraganglioma, a tumor that has not been reported in sufficient detail in previous literature. The patient was a 55-year-old woman with hypertension accompanied by an elevated serum norepinephrine level (2651 pg/mL; normal 100-450 pg/mL). Computed tomography revealed a well-circumscribed solid mass, 3.5 cm in diameter, located in the lower lobe of the left lung. In the lobectomy specimen, the tumor had invaded the B8 bronchus and hilar lymph nodes with microscopic metastasis to the mediastinal nodes. The tumor showed histologic, immunohistochemical, and ultrastructural features of paraganglioma: argyrophilic cells arranged in a nesting (Zellballen) or anastomosing trabecular pattern within an arcuate vascular network. Neoplastic chief cells positive for neuroendocrine markers (CD56, synaptophysin, chromogranin A) were surrounded by sustentacular cells positive for S-100 protein. Neurofilament protein was positively stained, but cytokeratins were totally negative. On electron microscopy, chief cells possessed abundant dense core granules with an eccentric halo ("norepinephrine-type" granules). The patient's blood pressure began to decline soon after the resection, and her serum norepinephrine promptly returned to almost normal. On the basis of our experience, our case is a bona fide primary pulmonary paraganglioma, a tumor heretofore subject to considerable skepticism.

Topics: CD56 Antigen; Chromogranin A; Chromogranins; Cytoplasmic Granules; Female; Humans; Hypertension; Keratins; Lung Neoplasms; Lymphatic Metastasis; Middle Aged; Neurofilament Proteins; Norepinephrine; Paraganglioma; S100 Proteins; Synaptophysin

Induced sputum, in contrast to bronchoscopic biopsies and lavages, is an easily obtained source of biological specimens. However, obtaining abnormal exfoliated cells for detailed molecular studies is limited because respiratory epithelial cells comprise only about 1% of sputum cell populations.. We developed a multiparameter flow sorting strategy to purify epithelial cells from nonepithelial sputum cells, using anti-cytokeratin antibody AE1/AE3 to recognize human epithelial cells and DAPI to stain DNA. We excluded cells with a high degree of side-scatter, which were composed predominantly of squamous cells and contaminating macrophages. The remaining cytokeratin-positive respiratory epithelial cells were then sorted based on anti-cytokeratin (PE) vs DNA (DAPI) parameters.. In this proof of principle study, the AE1AE3 cytokeratin/DNA flow sorting strategy enriched rare diploid respiratory epithelial cells from an average of 1.1% of cells in unsorted induced sputum samples to average purities of 42%. Thus, AE1AE3 flow-sorting results in a 38-fold enrichment of these cells.. We report a multiparameter flow cytometric assay to detect and enrich rare respiratory epithelial cells from induced sputum samples to average purities of 42%. With further development, this methodology may be useful as part of a molecular screening approach of populations at high risk for lung cancer.

Topics: Carcinoma; Cell Separation; Epithelial Cells; Flow Cytometry; Humans; Keratins; Lung Neoplasms; Sputum

Topics: Adenocarcinoma; Biomarkers, Tumor; Brain; Brain Neoplasms; Cerebrospinal Fluid; Diagnosis, Differential; Facial Nerve; Humans; Keratin-7; Keratins; Lateral Ventricles; Lung; Lung Neoplasms; Male; Meningeal Neoplasms; Middle Aged; Miller Fisher Syndrome; Oculomotor Nerve

To assess cytokeratin (CK) and thyroid transcription factor (TTF)-1 expression in primary epithelial lung tumours by comparison with non-pulmonary carcinomas and to correlate it with their histological type and grade.. Immunohistochemistry using antibodies against CKs 5/6, 7, 19, 20 and TTF-1 was applied to 165 primary and 37 secondary epithelial lung tumours. CK5/6 is a sensitive and specific marker of lung squamous carcinomas being positive in 100% of cases. CK7 is a common marker of primary lung adenocarcinomas (100% of cases) but with a lower specificity since it is also observed in other primary lung carcinomas (70% of large-cell neuroendocrine carcinomas, 40% of large-cell carcinomas, 23% of squamous carcinomas) but also in 27% of non-pulmonary adenocarcinomas. Addition of an anti-CK20 may be useful to prove or disprove the pulmonary origin of an adenocarcinoma when there is a history of colon cancer. CK19 is ubiquitous but a predominant or exclusive 'dot-like' pattern is very suggestive of high-grade neuroendocrine carcinoma. TTF-1 is a very sensitive and specific marker to document the pulmonary origin of an adenocarcinoma if a thyroid origin is excluded. Its expression in neuroendocrine lung tumours depends on the tumour grade.. Immunohistochemical expression of CKs and TTF-1 may be correlated with histological type and grade of lung primary epithelial tumours and may allow them to be distinguished from non-pulmonary carcinomas.

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

Based on the metastatic route in lymph nodes from lung cancer, we investigated micrometastasis in the dissected lymph nodes by genetic analysis of keratin 19 and nm23-H1 (the expression of a tumor-metastatic suppressor gene) and evaluated the postoperative outcomes.. Ten patients operated with lung cancer were 4 males and 6 females, who were stage IA; 2, stage IB; 3, stage IIA; 2, stage IIB; 1, and stage IIIA; 2, respectively. After total RNA extraction from the dissected lymph nodes, the expression of nm23-H1 and keratin 19 messenger ribonucleic acid (mRNA) were analyzed with reverse-transcripted polymerase chain reaction (RT-PCR).. The confirmation of micrometastasis in lymph nodes was realized in seven of 10 cases (70%), in their 5-year follow-up term. In three patients there was recurrence (43%, 3/7), and the one of them had died from the mediastinal recurrence. On the expression of nm23-H1 mRNA in lymph nodes, there was no significant difference between the pathologically lymph-node metastasis positive group and the negative one, and between the group with a tumor size over 30 mm and the group with a tumor size under 30 mm, respectively. The expression ratio of nm23-H1 gene was significantly expressed in the group with micrometastasis in lymph nodes (47%, 9/19) as compared to those without micrometastasis (10%, 1/10) (p<0.05). On the all-positive expression of nm23-H1 in the examined lymph nodes (n=4), no patient had recurrence (0%, 0/4). However, in the rest of the six patients without the all-positive expression of nm23-H1 in those lymph nodes (n=6), four patients had recurrence (67%, 4/6). There was no significance between the recurrent ratio in the all-positive expression of nm23-H1 suggesting lower incidence as compared to that in patients without all-positive expression of nm23-H1.. A detection of micrometastasis in lymph nodes could be a useful tool to identify the subpopulation of patients who might have a higher risk of recurrence and distant metastases. The nm23-H1 gene might be involved in a suppression role for micrometastasis in lymph nodes through the lymphatic route in lung cancer.

Topics: Aged; Chi-Square Distribution; Electrophoresis, Agar Gel; Female; Humans; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; NM23 Nucleoside Diphosphate Kinases; Nucleoside-Diphosphate Kinase; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

To investigate the prognostic factors in non-small cell lung cancer (NSCLC) at stage III and IV and establish a reliable model of clinical prognostic index.. Kaplan-Meier and Cox regression were used to analyze the relationship between the prognostic factors and survival time in 114 cases of NSCLC. The prognostic factors included clinical-pathological features and serum levels of cytokeratin fragment 19 (Cyfra21-1), CEA, neuron-specific enolase (NSE), CA125, interleukin-2 (IL-2) and soluble interleukin-2 receptors (sIL-2R).. Kaplan-Meier analysis showed that KPS, sex, disease stage, treatment, Cyfra21-1, sIL-2R and CA125 were related to prognosis. Multivariate analysis indicated that Cyfra21-1, stage and treatment were independent prognostic factors. When Cyfra21-1 > 3.5 mg/L, stage IV and chemotherapy < 3 cycles, the relative risk (RR) was 1.691, 2.229 and 3.035, respectively. In patients given 3 or more cycles of chemotherapy, serum Cyfra21-1, sIL-2R and stage at diagnosis were significantly independent prognostic factors. Three of these prognostic factors were used to establish a prognostic index (PI) model based on a simple algorithm: PI = Cyfra21-1 + sIL-2R + stage. The median survival period of patients with 3 or more cycles of chemotherapy were 18 months if PI = 0, 8 months if PI = 1 or 2, and 5 months if PI = 3.. The serum Cyfra21-1, sIL-2R and disease stage in unresectable NSCLC were independent prognostic factors. PI calculated on the basis of Cyfra21-1, sIL-2R and stage is recommended to predict the survival period of NSCLC.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Follow-Up Studies; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Prognosis; Proportional Hazards Models; Receptors, Interleukin-2; Survival Rate

To investigate the expression of CEA mRNA and CK(19) mRNA in peripheral blood cells of patients with lung cancer and evaluate its clinical significance.. Peripheral blood nucleated cells of 50 patients with lung cancer were studied by RT-PCR to detect the expression of CEA mRNA and CK(19) mRNA.. The combined positive rate of CEA mRNA and CK(19) mRNA in patients with lung cancer (82.0%) was significantly higher than that in patients with benign lung diseases (28.0%) and healthy volunteers (8.1%) (P < 0.001). The expression rate had no relation to the clinical staging or histological type. Compared with single detection, combined detection increased the detection rate but did not decrease the specificity.. Combined detection of CEA mRNA and CK(19) mRNA expression in peripheral blood nucleated cells increase the sensitivity of detecting hematogenous dissemination of cancer cells. Long-term survival analysis and more specimens would be helpful for evaluating its clinical significance.

Topics: Adenocarcinoma; Adult; Aged; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Topics: Aged; Aged, 80 and over; Asbestos; Biomarkers; Biomarkers, Tumor; Calbindin 2; Carcinoembryonic Antigen; Carcinoma, Small Cell; CD56 Antigen; Fatal Outcome; Histocytochemistry; Humans; Keratin-5; Keratins; Lung Neoplasms; Male; Mesothelioma; Neoplasms, Multiple Primary; Nuclear Proteins; Occupational Exposure; Pleural Neoplasms; S100 Calcium Binding Protein G; Thyroid Nuclear Factor 1; Transcription Factors; Vimentin

Two llamas with pulmonary tumors were examined. Llama No. 1 had multiple nodules throughout the lung that consisted histologically of solid clusters of polygonal to spindle cells with rare glandular differentiation. Intravascular emboli were common. Similar neoplastic masses were present in the kidney, heart, and liver. Immunohistochemically, neoplastic cells were positive for broad-spectrum cytokeratins (CKs), high-molecular weight CKs, CKs 5/6, and vimentin. The diagnosis was pulmonary carcinoma. Llama No. 2 had pulmonary nodules without extrapulmonary involvement. Microscopically, neoplastic cells formed acini lined by simple epithelium and solid cords of squamous cells that sometimes surrounded acini. Neoplastic cells were strongly positive for broad-spectrum CKs and weakly positive for thyroid transcription factor-1. The diagnosis was adenosquamous carcinoma. Pulmonary tumors account for 23% of neoplasms in South American camelids in our laboratory, making this the second most common type of neoplasm after lymphosarcoma.

Topics: Animals; Camelids, New World; Carcinoma, Adenosquamous; Diagnosis, Differential; Fatal Outcome; Female; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Vimentin

We studied the diagnostic value of Cdx2 to distinguish mucinous bronchioloalveolar carcinoma from mucinous colorectal adenocarcinoma metastatic to the lung. We retrieved 92 via the hospital computer system, including 30 mucinous bronchioloalveolar carcinomas, 32 nonmucinous bronchioloalveolar carcinomas, and 30 mucinous colorectal adenocarcinomas metastatic to the lung. All cases were confirmed by clinical history and surgical resection with occasional immunohistochemical studies. Cases were stained with antibodies against Cdx2, thyroid transcription factor-1 (TTF-1), cytokeratin (CK) 7, and CK20. Bronchioloalveolar carcinoma, mucinous type, showed positive staining for Cdx2, TTF-1, CK7, and CK20 in 0 (0%), 5 (17%), 30 (100%), and 18 (60%) of 30 cases, respectively; nonmucinous tumors were positive in 0 (0%), 30 (94%), 32 (100%), and 0 (0%) of 32 cases, respectively. For colorectal adenocarcinoma, the positive staining for Cdx-2, TTF-1, CK7, and CK20 was 29 (97%), 0 (0%), 7 (23%), and 29 (97%) of 30 cases, respectively. Our results demonstrated Cdx2 as a sensitive and specific marker for differentiating metastatic colorectal adenocarcinoma from mucinous bronchioloalveolar adenocarcinoma.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Avian Proteins; Biomarkers, Tumor; Colorectal Neoplasms; Diagnosis, Differential; Homeodomain Proteins; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

Recently, the authors identified molecular signatures and pathways associated with nonsmall cell lung carcinoma histology and lung development. They hypothesized that genetic classifiers of histology would provide insight into lung tumorigenesis and would be associated with clinical outcome when evaluated in a broader set of specimens.. Associations between patient survival and immunostaining for 11 representative histologic classifiers (epidermal growth factor receptor [EGFR], CDK4, syndecan-1, singed-like, TTF-1, keratin 5, HDAC2, docking protein 1, integrin alpha3, P63, and cyclin D1) were examined using a tissue microarray constructed from nonsmall cell lung carcinoma specimens.. Sixty-three tumors were examined, including 43 adenocarcinomas, 11 large cell carcinomas, and 9 squamous cell carcinomas. Sixty-three percent of tumors were clinical Stage I lesions, and 37% were Stage II-III lesions. In a multivariate analysis that controlled for age, gender, and race, syndecan-1 expression was found to be associated with a significant reduction in the risk of death (hazard ratio, 0.31 [95% confidence interval, 0.18-0.87]; P < 0.05). Multivariate analysis also indicated that EGFR expression was associated with a significant reduced risk of death.. The authors demonstrated that expression of either of the nonsmall cell lung carcinoma subtype classifiers syndecan-1 and EGFR was associated with a 30% reduction in the risk of death, with this reduction being independent of histology and other confounders. The results of the current study suggest that loss of expression of these histologic classifiers is associated with biologic aggressiveness in lung tumors and with poor outcome for patients with such tumors. If their significance can be validated prospectively, these biomarkers may be used to guide therapeutic planning for patients with nonsmall cell lung carcinoma.

Topics: Adenocarcinoma; Aged; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinases; ErbB Receptors; Female; Histone Deacetylase 2; Histone Deacetylases; Humans; Immunohistochemistry; Integrin alpha Chains; Keratin-5; Keratins; Lung Neoplasms; Male; Membrane Glycoproteins; Membrane Proteins; Multivariate Analysis; Nuclear Proteins; Proteoglycans; Proto-Oncogene Proteins; Repressor Proteins; Syndecan-1; Syndecans; Thyroid Nuclear Factor 1; Transcription Factors

We investigated the potential of circulating, nucleosomal DNA for the early prediction of the efficacy of chemotherapy in patients with advanced lung cancer.. In serum of 212 patients with newly diagnosed non-small cell lung cancer (stages III and IV) undergoing chemotherapy, nucleosomes (ELISA, Roche) were measured at days 1, 3, 5, and 8 of the first cycle and before each new therapeutic cycle. Additionally, carcinoembryonic antigen and cytokeratin 19 fragments (CYFRA 21-1; Elecsys, Roche) were determined before each cycle. The therapeutic success was classified by computed tomography before start of the third cycle according to the World Health Organization criteria.. In univariate analysis, responders (patients with remission) showed significantly (P < 0.05) lower values for the area under the curve of days 1 to 8 (AUC 1-8) of nucleosomes, the pretherapeutic baseline values of cycle 2 (BV2) and cycle 3 (BV3) of nucleosomes, and higher decreases of the baseline values from cycle 1 to 2 (BV1-2) and from cycle 1 to 3 (BV1-3) compared with nonresponders (patients with stable or progressive disease). Additionally, CYFRA 21-1 (BV1, BV2, BV3, BV1-2, BV1-3) and carcinoembryonic antigen (BV1-2) discriminated significantly between both groups. In multivariate analysis including all parameters available until end of the first therapeutic cycle, nucleosomes (AUC 1-8), CYFRA 21-1 (BV1), stage, and age were independent predictors of therapy response with nucleosomes (AUC 1-8) having the strongest impact.. Circulating nucleosomes in combination with oncological biomarkers are valuable for the early estimation of the efficacy of chemotherapy in patients with lung cancer.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antineoplastic Agents; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Drug Resistance, Neoplasm; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratin-19; Keratins; Kinetics; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Nucleosomes; Time Factors; Treatment Outcome

Lung cancer and pancreatic cancer are the most lethal tobacco-associated malignancies. To elucidate possible clinical interrelationships, the authors reviewed the clinicopathologic characteristics of patients treated for both pulmonary and pancreatic neoplasms.. Patients presenting with a potentially resectable pancreatic mass and a diagnosis of metachronous malignant neoplasm of the lung were studied by retrospective chart audit and review of histopathologic material.. Seven patients were identified over 6 years, representing five different clinical entities: metachronous presence of lung cancer and pancreatic cancer (n = 3), lung cancer metastatic to the pancreas (n = 1), lung cancer with a benign pancreatic neoplasm (n = 1), periampullary cancer metastatic to the lung (n = 1), and malignant melanoma metastatic to both lung and pancreas (n = 1). A tobacco history was present in all patients but one. Primary treatment modality was complete resection of isolated sites whenever feasible (lung resection, n = 6; pancreatic resection, n = 5). In four cases, a differential diagnosis of adenocarcinomas of both lung and pancreas was obtained after cytokeratin (CK) 7 and CK 20 immunohistochemistry. All patients with evidence of nodal or visceral metastasis from either primary site (n = 4) died within 5 to 9 months after the last operation. Three of four patients who had undergone resection of both pulmonary and pancreatic tumors were alive between 17 and 67 months after the last operation. All three survivors had presented with early disease stages and/or a protracted course (diagnostic interval, 16-66 months).. Our experience with neoplastic conditions that can involve lungs and pancreas metachronously may be useful to the clinician who is confronted with a similar situation. If therapeutic decision-making depends on differential diagnostic analysis, examination of CK 20 expression appears to be helpful. Although biologically favorable circumstances are rarely present, long-term survival seems possible after complete operative treatment in selected patients with early-stage disease.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Female; Humans; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Pancreatic Neoplasms; Postoperative Period; Retrospective Studies; Smoking

Histopathologic classification of lung carcinoma is important, as a prognostic factor and in the evaluation of treatment modalities. Although the World Health Organization classification of lung cancer is based on routine microscopy, immunohistochemistry is an important additional aspect in modern pathologic practice. This study examines whether the main histologic types of lung carcinomas are more reliably diagnosed with immunohistochemical technique using antibodies for the lung tissue-specific antigen thyroid transcription factor-1 (TTF-1) and a panel of cytokeratin (CK) antibodies. Forty-five cases of lung cancer (12 squamous cell carcinoma, 13 small cell carcinoma, 11 adenocarcinoma, 9 large cell carcinoma [LCC]/pleomorphic carcinoma) were stained with antibodies to CK CAM5.2, CK5, CK7, CK20, and TTF-1. All 45 cases were positive with CAM5.2, 16 of 45 cases with CK5, 34 of 45 cases with CK7, 4 of 45 cases with CK20, and 29 of 45 with TTF-1. Squamous cell carcinoma (epidermoid carcinoma) had the immunophenotype CK5+/TTF-1-, and at least 20% were also positive with CK7. Most nonepidermoid tumors had the "lung-specific" phenotype CK5-/TTF-1+; all small cell carcinomas had the phenotype CK5-/CK8+/TTF-1+, all adenocarcinomas CK5-/CK7+/TTF-1+ and (more than 50%) of LCC CK5-/CK7+/TTF-1+. Thus, more than 50% of LCCs were of the same phenotype as adenocarcinomas. The immunophenotypes of the main histologic types of lung carcinoma are stable and highly reproducible. However, because of considerable overlapping, immunophenotyping should not be used alone for histopathologic classification of lung cancer, but only as an adjunct to light microscopy. It is also suggested that CK5+ lung carcinomas with basaloid features should be regarded as variants of squamous cell carcinoma and not as LCC.

Topics: Biomarkers, Tumor; Carcinoma; Humans; Immunoenzyme Techniques; Immunophenotyping; Keratins; Lung Neoplasms; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

Numerous studies have been performed to determine diagnostic or prognostic utility of tumor markers in patients with lung cancer. The aim of the study was to evaluate the diagnostic usefulness of the tumor markers CA 125, CEA and CYFRA 21-1 in bronchoalveolar lavage fluid (BALF) in patients with non-small cell lung cancer (NSCLC). BAL was performed in 13 patients with NSCLC during diagnostic bronchofibroscopy. The control group consisted of 12 patients with sarcoidosis and 13 healthy volunteers. Tumor markers were determined in BALF supernatants using electrochemiluminescence technique (Elecsys 1010, Roche). To determine optimal cut-off values of tumor markers in BALF ROC curve was used. CEA and CA 125 concentration in BALF were significantly higher in NSCLC patients than in healthy volunteers and patients with sarcoidosis. CYFRA 21-1 in BALF was higher in NSCLC patients than in healthy volunteers, but no significant difference was found between NSCLC and sarcoidosis patients. The cut-off values of BALF concentration of CA 125, CEA and CYFRA 21-1 were 95 IU/mL, 3 ng/ml and 3 ng/ml, respectively. The sensitivity and specificity of CEA and CA 125 in BALF were 100%, 84% and 92%, 80%, respectively. In conclusion, we suggest that among the chosen markers, determination of CEA in BALF is the most useful in diagnosis of NSCLC. It may be a complementary method in diagnosing of patients in whom tumor cannot be visualized by bronchofibroscopy. These results need confirmation in larger groups of patients.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Bronchoalveolar Lavage Fluid; CA-125 Antigen; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Case-Control Studies; Female; Humans; Keratin-19; Keratins; Luminescent Measurements; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Sarcoidosis; Sensitivity and Specificity; Time Factors

To determine whether a micropapillary component is a prognostic predictor, with particular reference to nodal micrometastasis, in patients with stage I lung adenocarcinomas.. Thirty-five cases with stage I lung adenocarcinomas, obtained from lobectomies or pneumonectomies, and 434 dissected hilar and mediastinal lymph nodes, were retrospectively reviewed. A micropapillary component and nodal micrometastasis were found in 16 (45.7%) and 14 (40%) of the 35 cases, respectively, with nodal micrometastasis in 24 (5.5%) of the 434 lymph nodes, in an immunohistochemical study using an anti-cytokeratin antibody. Ten (62.5%) of the 16 cases with a micropapillary component, and four (21.1%) of the remaining 19 cases, showed nodal micrometastases (P = 0.014). Kaplan-Meier survival curves demonstrated that there was no significant difference between the cases with and without a micropapillary component (P = 0.28). However, the 5 years' survival of the cases with and without nodal micrometastases were 71.4% and 35.7%, respectively (P = 0.03).. A micropapillary component may be a manifestation of aggressive behaviour, as shown by frequent micrometastasis, for stage I lung adenocarcinomas.

Topics: Adenocarcinoma; Adult; Aged; Carcinoma, Papillary; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Retrospective Studies; Survival Analysis

We selected a 4-stain immunopanel including thyroid transcription factor (7ITF)-], cytokeratin (CK)7, 34betaE12, and CD56/neural cell adhesion molecule(NCAM) to subclassify a series of 45 pulmonary large cell carcinomas (LCCs) on bronchial biopsy. All cases consisted of a large tumor cell proliferation with abundant cytoplasm, vesicular nuclei, and prominent nucleoli. Immunohistochemically, 27 tumors (60%)were subclassified as adenocarcinoma (7TF-1 +/CK7+,24; CK7+ only, 3), 10 (22%) as squamous cell carcinoma (34betaE12+ only), and 4 (9%) as LCC with neuroendocrine differentiation (CD56+, variably stained with TTF-I and CK7, 34betaE12-). In 4 cases, the tumors coexpressed CK7 and 34betaE12 (3 cases) or were completely unstained (I case). Surgically resected tumors matched exactly with the corresponding original biopsy specimens in 21 of 23 cases; consistent CD56 expression was a reliable marker in confirming a diagnosis of large cell neuroendocrine carcinoma even on biopsy. Our results suggest that the proposed 4-stainset of commercially available markers might help subclassify LCC even in small biopsy material, validating expression-profiling studies aimed at lung cancer classification and permitting more consistent patient enrollment for trials with targeted treatments.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Large Cell; Carcinoma, Squamous Cell; CD56 Antigen; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Neuroendocrine Tumors; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

Serum tumor markers are non-invasive diagnostic tools for malignant tumor and commonly used for screening of cancer and as an indicator of treatment-effect. In small cell lung cancer, NSE and proGRP are effective markers. In non-small cell lung cancer (NSCLC), CEA, SCC, CYFRA21-1, SLX and CA19-9 are commonly used for screening, and at least one marker among CEA, SCC or CYFRA21-1 is positive in 77% of patients with NSCLC. According to the histological type, the positive rate of CEA and CYFRA21-1 is high in adenocarcinoma patients, and the positive rate of CYFRA21-1 and SCC is high in squamous cell carcinoma patients. This review summarizes the clinical usefulness of tumor markers in primary lung cancer.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Humans; Keratin-19; Keratins; Lewis X Antigen; Lung Neoplasms; Peptide Fragments; Peptides; Recombinant Proteins; Serpins

A large primary retroperitoneal sublumbar neoplasm was identified in an 11-year-old Holstein cow, with metastases to the lungs, kidneys and lymph nodes. The tumour cells proliferated in a characteristic endocrine pattern, were argyrophilic and positive for neuron-specific enolase, and had membrane-bounded intracytoplasmic granules. In addition, the cells were occasionally positive for cytokeratin and had desmosome-like intercellular junctions. The primary tumour mass was diagnosed as a malignant paraganglial tumour of the aortico-sympathetic ganglion (organ of Zuckerkandl), and was considered to contain primitive cells with epithelial differentiation.

Topics: Animals; Biomarkers, Tumor; Cattle; Cattle Diseases; Cytoplasmic Granules; Desmosomes; Female; Ganglia, Sympathetic; Immunoenzyme Techniques; Keratins; Kidney Neoplasms; Lung Neoplasms; Lymph Nodes; Paraganglioma; Phosphopyruvate Hydratase; Retroperitoneal Neoplasms; Silver Staining

Metastatic cancer in thoracic lymph nodes without a primary site is rare. The purpose of this study is to draw attention to this probably underestimated entity, to speculate on its possible origins, and to suggest guidelines for its treatment.. Eight heavy smokers with no past medical history of cancer were diagnosed at operation to have malignant cells in intrathoracic lymph nodes (N1 or N2) with no primary site in the lung. All patients underwent an exploratory thoracotomy with a presumed diagnosis of lung cancer except one who presented with a middle lobe mucosa-associated lymphoid tissue lymphoma. We reviewed the type of surgical resection, histologic and immunohistochemical analysis of resected specimens, treatments, survival, and long-term results.. Resections performed were pneumonectomy (n = 4), lobectomy (n = 3), and bilobectomy (n = 1). All patients underwent complete mediastinal lymph node dissection. Lung resection was performed for mucosa-associated lymphoid tissue lymphoma (n = 1) and for tumorlike lesions that appeared to be tuberculoma (n = 1) and intrapulmonary metastatic lymph nodes (n = 6). Malignant cells were located in intrapulmonary lymph nodes alone (n = 3) or also in mediastinal lymph nodes in three other cases. All these tumors were cytokeratin-positive, demonstrating their epithelial nature. Pulmonary origin was confirmed in two cases (thyroid transcription factor 1-positive and thyroglobulin-negative). No other origin could be demonstrated by immunochemistry. Three patients died within the first year. All other patients are still alive without recurrence (Kaplan-Meier 5-year survival rate, 62.5%).. Frequency of metastatic cancer in thoracic lymph nodes without a primary site is probably underestimated because the cancer is routinely diagnosed by mediastinoscopy and considered as metastatic disease not amenable to operation. The origin of the disease, either pulmonary, endogenous, or from extrathoracic sites, is often difficult to assess. Nevertheless, our data confirm those of the literature and demonstrate that survival can be increased by operation. This implies diagnosis of the entity and consideration that thoracic lymph node involvement can apparently be isolated and therefore resectable.

Topics: Aged; Histocytochemistry; Humans; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Lymphoma, B-Cell, Marginal Zone; Middle Aged; Neoplasms, Unknown Primary; Nuclear Proteins; Pneumonectomy; Thyroglobulin; Thyroid Nuclear Factor 1; Transcription Factors; Tuberculoma

Several immunohistochemical markers, among them calretinin, thrombomodulin (CD141), keratin 5, and mesothelin, have been documented or suggested as useful markers for positive identification of mesothelioma and to differentiate it from pulmonary adenocarcinoma; numerous studies have documented their variable specificity. However, expression of these markers in other types of lung carcinomas has not been systematically explored, although these tumors can enter in the differential diagnosis of mesothelioma. In this study we immunohistochemically evaluated 596 lung carcinomas of different types for the four above-mentioned mesothelioma markers, all of which reacted with a great majority of epithelioid mesotheliomas studied for comparison. Calretinin expression was common in giant cell carcinomas (67%), small cell carcinomas (49%), and large cell carcinomas (38%), whereas it was rare in usual adenocarcinomas but slightly more common in those with neuroendocrine differentiation (11% and 17%, respectively). Thrombomodulin was present in all keratinizing squamous carcinomas and the great majority (87%) of nonkeratinizing tumors in a membrane-staining pattern. It was moderately common in small cell (27%) and large cell carcinomas (25%) but relatively rare in adenocarcinomas (13%). Keratin 5 was expressed in all keratinizing and the great majority (87%) of nonkeratinizing squamous carcinomas, and a majority of large cell carcinomas (56%) and some small cell carcinomas (27%). It was rare in acinar adenocarcinomas (12%) and absent in those with neuroendocrine differentiation. Mesothelin was present in more than half (53%) of adenocarcinomas and a minority (13%) of large cell carcinomas but was absent in small cell carcinomas. In squamous carcinomas it was more often seen in nonkeratinizing versus keratinizing tumors (31% vs 16%). These results show that each of these "mesothelioma" markers reacts with different subsets of pulmonary carcinomas with a variable frequency; this should be considered when using these markers in the differential diagnosis of thoracic tumors.

Topics: Biomarkers, Tumor; Calbindin 2; Carcinoma; Diagnosis, Differential; Humans; Immunohistochemistry; Keratin-5; Keratins; Lung Neoplasms; Mesothelioma; Pleural Neoplasms; S100 Calcium Binding Protein G; Thrombomodulin

Monoclonal antibody 34betaE12 (Ck34betaE12) recognizes a set of cytokeratins (1, 5, 10, 14) expressed in normal stratified squamous epithelium. We have recently reported its expression in squamous cell carcinoma and basaloid carcinoma, in contrast to large cell neuroendocrine carcinoma, an entity with overlapping morphological features with basaloid carcinoma. We have now examined the role of Ck34betaE12 in discriminating between neuroendocrine and non-neuroendocrine proliferations.. We performed an immunohistochemical study of 228 cases, comprising the whole spectrum of lung neuroendocrine proliferations and tumours. All cases of neuroendocrine cell hyperplasia (n = 15), tumorlet (n = 23), typical carcinoid (n = 27) and atypical carcinoid (n = 23) were completely negative for Ck34betaE12. Although the neuroendocrine cells of small cell lung carcinoma and large cell neuroendocrine carcinoma were consistently negative, a strong and diffuse positive staining was found in the non-neuroendocrine components of combined small cell carcinoma (three of eight cases) and combined large cell neuroendocrine carcinoma (11 of 12 cases). In addition, scattered Ck34betaE12+ cells were noted in 11 of 64 (17%) large cell neuroendocrine carcinoma and in seven of 56 (12.5%) small cell carcinoma, which were not obviously histologically combined. This heterogeneity of high-grade neuroendocrine tumours was not observed in carcinoids which lack Ck34betaE12 clusters of reactive cells. There was mutual exclusion between expression of neuroendocrine markers and that of Ck34betaE12.. We conclude that 34betaE12 expression excludes the neuroendocrine nature of tumour cells and uncovers the real frequency of combined forms in high-grade neuroendocrine tumours.

Topics: Biomarkers, Tumor; Carcinoma, Neuroendocrine; Carcinoma, Small Cell; Fluorescent Antibody Technique, Indirect; Humans; Hyperplasia; Immunoenzyme Techniques; Keratins; Lung Neoplasms

This retrospective study aimed at determining the prognostic significance of neuroendocrine markers chromogranin A (CgA), pro-gastrin releasing peptide (ProGRP) and neuron-specific enolase (NSE), together with the cytokeratin 19 marker CYFRA 21-1 in small cell lung cancer (SCLC). A total of 148 histologically proven and previously untreated SCLC patients were included. Among them 118 patients received a cisplatin-etoposide combination or cisplatin-etoposide-cyclophosphamide-4'-epidoxorubicin combination. All tumour markers were tested using immunoradiometric assays except for ProGRP which was tested using an enzyme-linked immunosorbent assay. The thresholds for marker serum titrations were 53 pg/ml, 65, 17, and 3.6 ng/ml for ProGRP, CgA, NSE and CYFRA 21-1 respectively. Univariate analysis showed that patients affected by one of the following characteristics proved to have a significant shorter survival in comparison with the opposite status of each variable: age over 63 years, extensive-stage, serum LDH level higher than 600 U/l, serum NSE level higher than 17 ng/ml, serum CgA level higher than 65 ng/ml and serum CYFRA 21-1 level higher than 3.6 ng/ml. In addition, there was a trend towards a statistical significance for a high serum alkaline phosphatase level and a performance status equal to or worse than two. The following variables were independent determinants of a poor outcome: a poor performance status (hazard ratio [95% confidence interval]: 1.51 [1.02-2.22]), a high CgA level (HR: 1.61 [1.06-2.45]), a high CYFRA 21-1 level (HR: 2.10 [1.40-3.14]) and an age older than 63 years (HR: 1.68 [1.14-2.48]). When the multivariate analysis was restricted to patients receiving a cisplatin-etoposide-based chemotherapy, the same variables were prognostic determinants with nearly similar hazard ratios. In conclusion, aside classical variables such as age and performance status, high serum CYFRA 21-1 and high serum CgA level in SCLC are both prognostic determinants of prognosis, in particular in patients receiving conventional chemotherapy consisting of cisplatin and etoposide-based combinations.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Small Cell; Chromogranin A; Chromogranins; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Peptide Fragments; Peptides; Phosphopyruvate Hydratase; Prognosis; Recombinant Proteins; Retrospective Studies; Survival Rate

We collected 75 primary pulmonary carcinomas with pleomorphic, sarcomatoid, or sarcomatous elements to better define their clinical, histologic, and immunohistochemical profile. The patient's age ranged from 42 to 81 years (mean 65 years), and the male-to-female ratio was 9.7:1. Sixty-nine patients (92%) were smokers. Cough and hemoptysis were the most frequent presenting symptoms. Fifty-nine patients (65%) died of disease: only stage significantly predicts overall survival (p = 0.0273). Microscopically, based on the WHO criteria, 58 cases were classified as pleomorphic carcinoma (51 with an epithelial component, 7 composed exclusively of spindle and giant cells), 10 as spindle cell carcinoma, 3 as giant cell carcinoma, 3 as carcinosarcoma, and 1 as pulmonary blastoma. Immunohistochemically, in the tumors composed exclusively of spindle and/or giant cells, thyroid transcription factor-1 (TTF-1) and cytokeratin 7 were positive in 55% and 70% of the cases, respectively, whereas surfactant protein-A was always negative. In pleomorphic carcinomas with an epithelial component, cytokeratin 7, TTF-1, and surfactant protein-A were positive in the sarcomatoid component in 62.7%, 43.1%, and 5.9% of the cases, respectively, whereas they were always negative in the sarcomatous part of carcinosarcomas and blastoma. In the epithelial component of pleomorphic carcinomas, cytokeratin 7, TTF-1, and surfactant protein-A were positive in 76.4%, 58.8%, and 39.2% of the cases, respectively, whereas the same antibodies did not react with the epithelial component of carcinosarcomas; in the case of blastoma, the epithelial part of the tumor was positive for cytokeratin 7 and TTF-1, whereas it was negative for surfactant protein-A. Cytokeratin 20 was always negative. In our opinion, this study: 1) supports the metaplastic histogenetic theory for this group of tumors; 2) shows that cytokeratin 7 and TTF-1, but not surfactant protein-A, are useful immunohistochemical markers in this setting; 3) confirms that stage is at the moment the only significant prognostic parameter, as in conventional non-small cell lung carcinomas; and 4) shows that this group of tumors has a worse prognosis than conventional non-small cell lung carcinoma at surgically curable stages I, justifying their segregation as an independent histologic type in the WHO classification.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma; Carcinosarcoma; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Prognosis; Pulmonary Surfactant-Associated Protein A; Survival Rate; Thyroid Nuclear Factor 1; Transcription Factors

Rates of eukaryotic protein synthesis and proliferation are dependent upon the availability of eIF4F, the cap-binding translation initiation complex that guides the ribosome onto the mRNA. One possible rate-limiting factor in eIF4F complex formation is the availability of eIF4E, which interacts specifically with the mRNA cap structure. As such, it has a potential role in the selective translation of growth-related mRNAs, with overexpression of eIF4E resulting in aberrant cell growth and transformation. A number of studies suggest that eIF4E may play a role in cellular differentiation as well as proliferation. We have previously reported that post-transcriptional regulation is involved in the induction of keratins in epithelial lung tumor cell lines exposed to the differentiation-modulating agent, bromo-deoxyuridine (BrdU). Here, we demonstrate that these BrdU-treated lung cells express elevated levels of eIF4E protein and enhanced phosphorylation of eIF4E. Overexpression of eIF4E by cDNA transfection in the poorly differentiated, keratin-negative human lung cell line, DLKP, was found to promote a flattened, more epithelial appearance to these cells, coupled with the induction of simple keratins (keratins 8 and 18). In contrast, levels of eIF4E expression were found to decrease during BrdU-induced differentiation of the leukemic cell line, HL-60, suggesting that there are cell-type differences in the response to BrdU and in the requirement for eIF4E during differentiation.

Topics: Bromodeoxyuridine; Cell Differentiation; Epithelium; Eukaryotic Initiation Factor-4E; Humans; Keratin-8; Keratins; Lung Neoplasms

It has been reported previously in cases of adenosquamous carcinoma of the lung in Okinawa, a subtropical island 2000 km south of mainland Japan, that the squamous cell carcinoma components were positive for human papillomavirus (HPV) by non-isotopic in situ hybridisation (NISH). The adenocarcinoma cells adjacent to the squamous cell carcinoma components were enlarged and also positive for HPV. This is thought to indicate that after adenocarcinoma cells are infected with HPV, they undergo morphological changes, and that "squamous metaplasia" follows. In this present study, the effects of HPV transfection into adenocarcinoma cells were examined. The relation between the region expressing the HPV gene and squamous metaplasia was also studied.. Plasmid pBR322 containing HPV type 16 (HPV-16) was transfected into cultured colonic adenocarcinoma (DLD-1) and lung adenocarcinoma (PC-14) cells using the calcium phosphate method. Neomycin was used as a selection marker. The presence of HPV E1, E2, E4, E5, E6, E7, L1, and L2 mRNAs and also transglutaminase 1, involucrin, cyclin dependent kinases (CDKs), cyclins, caspases, apoptosis inducing factor, DNase gamma, Fas, and Fas ligand mRNAs in HPV transfected cells was investigated by means of reverse transcription polymerase chain reaction (RT-PCR). The G0-G1 cell population was analysed by flow cytometry. Morphological examination under light and electron microscopes was also carried out.. The virus transfected cells showed squamous metaplasia when they were injected into severe combined immunodeficient mice, expressing the high molecular weight keratin (Moll's number 1 keratin) and involucrin molecules immunohistochemically, and involucrin and transglutaminase I mRNAs by RT-PCR. The squamous metaplasia was most conspicuous in the HPV transfected DLD-1 cell when compared with HPV transfected PC-14 cells. Squamous metaplasia was most clearly demonstrated in one HPV transfected DLD-1 cell clone, which expressed not only E2 but also E6-E7 fusion gene mRNA. Viral L1 mRNA expression was absent in HPV transfected cell clones, and was not related to squamous metaplasia. The growth rate of HPV transfected cells was reduced. Transfection of the virus into the cultured adenocarcinoma cells increased the G0-G1 cell population greatly, as assessed by flow cytometer analysis. Furthermore, in the virus transfected cells, apoptosis was also observed by means of the terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labelling method.. HPV transfection into adenocarcinoma cells induced clear squamous metaplasia. One of the HPV transfected cell clones that expressed E2 and E6-E7 fusion gene mRNA showed the squamous metaplasia particularly clearly, and apoptosis was also demonstrated.

Topics: Adenocarcinoma; Animals; Apoptosis; Blotting, Western; Carcinoma, Squamous Cell; Cell Cycle; Cell Differentiation; DNA, Viral; Humans; Keratins; Lung Neoplasms; Metaplasia; Mice; Mice, SCID; Neoplasm Proteins; Neoplasm Transplantation; Papillomaviridae; Protein Precursors; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Viral; Transfection; Tumor Cells, Cultured

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Antineoplastic Agents; Biomarkers, Tumor; Fatal Outcome; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Middle Aged; Nuclear Proteins; Pneumonectomy; Thyroid Nuclear Factor 1; Transcription Factors

Carcinomas with micropapillary features have been described in the breast, urinary bladder, lung, and ovary. They are characterized by the presence of micropapillary tufts in clear spaces. Unequivocal vascular invasion is usually present at the periphery of the tumor. Consequently, these tumors have a high propensity for lymph node metastases and high-stage disease. The metastatic carcinoma can consist exclusively of the micropapillary component, which may elicit an erroneous diagnosis if located in the bladder or lung, as in the patient presented herein. We present a case of a 59-year-old woman with a history of bilateral breast carcinoma status post-bilateral mastectomy, chemotherapy, and tamoxifen therapy. She presented with urinary frequency, and a pelvic mass was noted. A biopsy of the endometrium revealed a poorly differentiated carcinoma. Urinary bladder biopsies showed a carcinoma with micropapillary features diagnosed as micropapillary transitional cell carcinoma. She presented to M.D. Anderson Cancer Center (Houston, TX) for further treatment recommendations. The urinary bladder and endometrial biopsies both contained carcinomas with micropapillary features. The mastectomy specimen showed an invasive ductal carcinoma with a significant micropapillary component. The tumor cells from the breast, endometrium, and urinary bladder were positive for cytokeratin (CK) 7 and estrogen receptor and negative for CK20. In view of the morphologic and immunohistochemical profile, the carcinoma in the endometrium and urinary bladder were interpreted as metastatic lesions from the breast primary. Carcinomas with a micropapillary component are morphologically identical in the breast, urinary bladder, and lung. However, micropapillary serous carcinoma has a different appearance more akin to borderline tumors of the ovary. Immunohistochemical stains are useful in distinguishing these lesions in that thyroid transcription factor-1 positivity suggests a lung primary, CK7 and estrogen receptor suggest a breast primary, and both CK7 and CK20 positivity suggest a urinary bladder primary. It is important to exclude metastatic carcinomas with micropapillary features before making a definite diagnosis of a primary tumor. Carcinomas with micropapillary features have a propensity for lymph node metastases and advanced stage disease. This article discusses the differential diagnosis of carcinomas with micropapillary features in different organs.

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Papillary; Carcinoma, Transitional Cell; Diagnosis, Differential; Endometrial Neoplasms; Female; Humans; Immunohistochemistry; Keratin-7; Keratins; Lung Neoplasms; Middle Aged; Receptors, Estrogen; Urinary Bladder Neoplasms

The objective was to elaborate a method of immunohistochemical diagnosis of bone micrometastases and to apply this procedure in operated patients with the non-small cell form of lung cancer. By means of a cocktail of cytokeratin mixtures and individual cytokeratins the immunohistochemical profile of the primary pulmonary focus is assessed. Bone micrometastases are evaluated from the bone marrow of a sampled costal segment. A total of 30 bone marrow samples were examined. For further evaluation 17 patients remained in the group. Isolated segments with a marked positivity of cytokeratins were observed in four instances. The yield of bone marrow from costal segment is very good. The initial results are encouraging for further research.

Topics: Biomarkers, Tumor; Bone Neoplasms; Carcinoma, Non-Small-Cell Lung; Humans; Immunohistochemistry; Keratins; Lung Neoplasms

Our objective was to test the prognostic importance of both the pretreatment level and change in serum CYFRA 21-1 after one cycle of chemotherapy in patients with advanced non-small cell lung cancer (NSCLC) and to compare these two CYFRA variables to routine clinical stage and response as measured by imaging.. Our patients consisted of 58 with advanced NSCLC who were treated with chemotherapy. Fourteen were stage IIIa, 8 stage IIIb, and 36 stage IV, and none had received previous treatment. The choice of chemotherapy was left to the discretion of the treating physicians. We collected two serum samples, one before the first cycle of chemotherapy and the second before the second cycle, and analyzed these for serum CYFRA 21-1 using an electrochemiluminescence immunoassay and the ElecSys 2010 system (Roche Diagnostics Corp., Indianapolis, IN). We expressed changes in CYFRA in terms of the natural ratio logarithm of post-treatment to pretreatment CYFRA, and we used the Cox proportional hazards model to analyze survival time.. Patients experienced an average drop of 27% in serum CYFRA after the first cycle of chemotherapy. Furthermore, the Cox model demonstrated that both the initial natural logarithm of serum CYFRA and presence of >27% drop in CYFRA were significantly related to subsequent survival (model P < 0.0006), but neither clinical stage nor clinical response related to survival (P > 0.1).. In advanced stage NSCLC, the initial level of serum CYFRA appears to provide more prognostic information than clinical stage. Furthermore, a drop of >27% in CYFRA after one cycle of therapy adds prognostic information, so that this threshold appears to be an early measure of response to chemotherapy.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Pilot Projects; Prognosis; Sensitivity and Specificity; Survival Rate

We reported previously a significant increase in survival of nude rats harboring orthotopic A549 human non-small cell lung cancer tumors after treatment with a combination of exisulind (Sulindac Sulfone) and docetaxel (D. C. Chan, Clin. Cancer Res., 8: 904-912, 2002). The purpose of the current study was to determine the biochemical mechanisms responsible for the increased survival by an analysis of the effects of both drugs on A549 orthotopic lung tumors and A549 cells in culture. Orthotopic A549 rat lung tissue sections from drug-treated rats and A549 cell culture responses to exisulind and docetaxel were compared using multiple apoptosis and proliferation analyses [i.e., terminal deoxynucleotidyl transferase-mediated nick end labeling, active caspase 3, the caspase cleavage products cytokeratin 18 and p85 poly(ADP-ribose) polymerase, and Ki-67]. Immunohistochemistry was used to determine cyclic GMP (cGMP) phosphodiesterase (PDE) expression in tumors. The cGMP PDE composition of cultured A549 cells was resolved by DEAE-Trisacryl M chromatography and the pharmacological sensitivity to exisulind, and additional known PDE inhibitors were determined by enzyme activity assays. Exisulind inhibited A549 cell cGMP hydrolysis and induced apoptosis of A549 cells grown in culture. PDE5 and 1 cGMP PDE gene family isoforms identified in cultured cells were highly expressed in orthotopic tumors. The in vivo apoptosis rates within the orthotopic tumors increased 7-8-fold in animals treated with the combination of exisulind and docetaxel. Exisulind increased the in vivo apoptosis rates as a single agent. Docetaxel, but not exisulind, decreased proliferative rates within the tumors. The data indicate that exisulind-induced apoptosis contributed significantly to the increased survival in rats treated with exisulind/docetaxel. The mechanism of exisulind-induced apoptosis involves inhibition of cGMP PDEs, and these results are consistent with a cGMP-regulated apoptosis pathway.

Topics: 3',5'-Cyclic-GMP Phosphodiesterases; Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Carcinoma, Non-Small-Cell Lung; Caspase 3; Caspases; Cell Division; Docetaxel; Female; Humans; In Situ Nick-End Labeling; Keratins; Ki-67 Antigen; Lung Neoplasms; Poly(ADP-ribose) Polymerases; Rats; Rats, Nude; Sulindac; Survival Rate; Taxoids; Tumor Cells, Cultured

Cytokeratin 19 fragment (CYFRA) is a specific tumor marker in patients with lung cancer; however, it has been reported that serum CYFRA levels are elevated in some patients with nonmalignant respiratory diseases such as interstitial pulmonary fibrosis (IPF) and collagen disease-associated pulmonary fibrosis (CDPF). To investigate the serum CYFRA levels in nonmalignant respiratory diseases in detail, we studied 413 patients with respiratory diseases.. Retrospective study.. University hospital.. Four hundred thirteen patients with nonmalignant respiratory diseases and lung cancer.. Serum CYFRA levels were measured with a commercially available enzyme immunoassay kit. Immunohistochemical study was performed using monoclonal antibody Ks 19.1 (Rosch Diagnosica; Bern, Switzerland) on surgically resected or autopsied lung specimens. Gel electrophoresis and immunoblotting was performed with serum samples. In 149 patients with nonmalignant diseases except IPF and CDPF, the ratio of patients with > 3.5 ng/mL of serum CYFRA was 13.4%. In 13 of 30 patients (43.3%) with IPF and CDPF, the serum CYFRA levels were abnormally elevated. The 95th percentile serum CYFRA level of the patients with nonmalignant respiratory diseases was 6.2 ng/mL, and none of them had CYFRA levels > 20.3 ng/mL. Survival in patients with IPF and CDPF with elevated serum CYFRA levels were significantly lower than in those with normal range (p = 0.0335). Western blotting using serum from nonmalignant lung diseases and patients with lung cancer showed no apparent difference between them. An immunohistochemical study indicated CYFRA was selectively expressed in the pulmonary epithelial cells that covered the remodeling alveolar septi in nonmalignant respiratory disease.. Serum CYFRA was elevated in some nonmalignant respiratory diseases, especially in IPF and CDPF. The value of serum CYFRA would reflect the severity of lung injury in nonmalignant respiratory diseases and might be related to the prognosis in patients with IPF and CDPF.

Topics: Adult; Biomarkers, Tumor; Collagen Diseases; Electrophoresis, Polyacrylamide Gel; Epithelial Cells; Female; Humans; Immunoblotting; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Male; Pulmonary Fibrosis; Respiratory Tract Diseases; Retrospective Studies

This study was designed to prospectively substantiate the prognostic value of cytokeratin-positive (CK(+)) cells in the bone marrow (BM) and regional lymph nodes (LNs) in resected nonsmall cell lung cancer (NSCLC) patients from a large population within a multicenter study.. The study population consisted of 351 patients with stages I to IIIA NSCLC from 15 Japanese institutes. BM aspirates were stained immunocytochemically with the anti-cytokeratin antibody, CK2. The hilar and mediastinal LNs of 216 patients with stage I NSCLC were stained immunohistochemically with the anti-CK antibody, AE1/AE3.. CK(+) cells were detected in 112 patients (31.9%) of the 351 BM aspirate patients. The frequency of CK(+) cells showed no differences among pathologic stages. The patients with CK(+) cells in the BM had a tendency to have shorter survival periods than those without CK(+) cells (p = 0.076). Although the presence of CK(+) cells in the BM of patients with stage I did not allow the prediction of overall survival, it reduced the overall survival significantly in patients with stages II to IIIA. CK(+) cells in the LNs were detected in 34 of 216 patients (15.7%) with stage I. The patients with CK(+) cells in the LNs had a poor prognosis by both univariate (p = 0.004) and multivariate analyses (p = 0.018).. The presence of CK(+) cells in the BM was related to a poor prognosis for patients with stages II to IIIA NSCLC; however, it did not predict the prognosis of patients with stage I. For stage I NSCLC, the detection of CK(+) cells in the LNs implied a poor prognosis for the patients.

Topics: Adult; Aged; Biomarkers, Tumor; Biopsy, Needle; Bone Marrow; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratins; Lung Neoplasms; Lymph Nodes; Male; Middle Aged; Neoplasm Staging; Pneumonectomy; Probability; Prognosis; Proportional Hazards Models; Prospective Studies; Sensitivity and Specificity; Survival Analysis; Treatment Outcome

Lung adenocarcinoma presenting as malignant pleural effusion (MPE) is common in Taiwan. Microscopically, the involved pleurae are infiltrated by numerous tumor foci, which suggests that the cancer cells are highly invasive. Overexpression of HER-2/neu has been related to proliferation, antiapoptosis, and the high invasiveness of various cancer cells. We therefore were interested in studying the role of HER-2/neu in MPE-associated adenocarcinoma cell lung cancer (ADCLC).. The expression of HER-2/neu in pleural effusion was measured by ELISA. The HER-2/neu protein expression on tumor cells was evaluated by immunohistochemical (IHC) staining, and gene amplification was assayed by fluorescence in situ hybridization.. The mean value of HER-2/neu in pleural effusions of patients with ADCLC and other nonmalignant lung diseases was 9.9 and 2.7 ng/ml, respectively. The difference is statistically significant (P < 0.001). Compared with cytokeratin 19 fragment CYFRA 21-1, the performance of HER-2/neu as a tumor marker in pleural effusion diagnosis was better. Overexpression of HER-2/neu in tumor tissues was found in 70% (23 of 32) of patients with MPE-associated ADCLC, 30% (13 of 43) with stage I/II non-small cell lung cancer (NSCLC), and 44% (14 of 32) with stage III NSCLC. The incidence of HER-2/neu overexpression in tumor tissues of patients with MPE-associated ADCLC was significantly higher than that of patients with stage I-III NSCLC without MPE. HER-2/neu gene amplification was uncommon (1.9%). The correlation between the IHC H-score in tumor samples and the pleural effusion level of HER-2/neu was significant (P < 0.01). A higher incidence of HER-2/neu expression beyond the cutoff point (5.5 ng/ml) in pleural effusions was also found in patients whose IHC H-scores were >50.. These findings indicate that HER-2/neu is important in the pathogenesis of MPE-associated ADCLC and is a potential tumor marker for a diagnosis of pleural effusion.

Topics: Adenocarcinoma; Antigens, Neoplasm; Apoptosis; Biomarkers, Tumor; Cell Division; Cell Line, Tumor; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratin-19; Keratins; Lung Neoplasms; Neoplasm Metastasis; Pleural Effusion; Receptor, ErbB-2

We have longstanding experience with tissue polypeptide antigen (TPA), a tumor marker of the cytokeratin (CK) family. In the mid-1990s, a new CK marker, CK 19 fragments (CYFRA 21-1), became popular and widely accepted. This is the first study specifically designed to compare the two markers.. Analysis of a single institution database over a 3-year period (ie, 1998 to 2000).. Community-based hospital and second referral level institution for a province of 500,000 people.. The study included 180 new consecutive patients (143 men) with pathologically documented non-small cell lung cancer (NSCLC), who were observed during and after treatment, and eventually were assessed for status.. Anthropometric, clinical, and laboratory data, including TPA and CYFRA 21-1 serum levels, were recorded prospectively. Standard nonparametric tests, Kaplan-Meyer survival analyses, Cox proportional hazards models, receiver-operating characteristic (ROC) curves, and estimates were used for statistical analysis.. A total of 1,299 twin TPA and CYFRA 21-1 serum assays (180 performed at diagnosis and 1,119 performed during or after treatment) were obtained. Intermarker correlation tests revealed incredibly high Spearman rho indexes, ranging from 0.935 at diagnosis to 0.813 to 0.921 at the different follow-up times. The substantial equivalence of the two tests explained all the other results, as follows: their similar profile of correlation with the other variables (objective treatment response: TPA rho, 0.456; CYFRA 21-1 rho, 0.463; follow-up performance status: rho range, 0.424 to 0.435); their superimposable capability to predict important clinical situations (eg, recognizing a metastatic disease at diagnosis with areas under the ROC curve of 0.742 and 0.706, respectively); their nearly identical prognostic significance (the D statistic of the goodness-of-fit of a multivariate survival model: TPA, 851.0; CYFRA 21-1, 851.6).. In most of their traditional clinical applications the two serum tests are equivalent because of their virtual identity. We strongly recommend using a CK test in the evaluation of each NSCLC patient. The choice between TPA and CYFRA 21-1 can be based on nonclinical factors, such as the laboratory experience or preference, and the cost of the two kits.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Prognosis; Prospective Studies; ROC Curve; Survival Analysis; Tissue Polypeptide Antigen

To evaluate the utility of thyroid transcription factor-1 (TTF-1), surfactant protein-B, (SP-B) cytokeratin 7 (CK7) and CK20 immunostaining in the discrimination between primary adenocarcinomas and metastatic adenocarcinomas.. Formalin fixed, paraffin embedded tissue blocks from 42 primary lung adenocarcinomas and 30 metastatic carcinomas resected during operation were immunostained with monoclonal antibodies to TTF-1, SP-B, CK7, and CK20.. Positive immunostaining with TTF-1 and SP-B was noted in 74% and 52% of primary lung tumor, respectively. The positive immunostaining and specificity of such a combination for discriminating between primary and metastatic adenocarcinoma were 79% and 94%, respectively. All primary lung adenocarcinomas were CK7 positive, 24 (57%) were CK7 positive/CK20 negative and 18 were CK7 positive/CK20 positive in immunophenotype. Colon and breast were the most common sites of metastasis. All metastatic colorectal adenocarcinomas were CK20 positive, 11 (92%) were CK7 negative/CK20 positive and 1 was CK7 positive/CK20 positive in immunophenotype. The results of cytokeratin immunostaining in the metastatic breast tubular carcinomas were similar to those in the primary lung adenocarcinomas: 4 were CK7 positive/CK20 negative and 4 were CK7 positive/CK20 positive. CK7 positive, and TTF-1 or SP-B positive immunophenotype was seen in 33 (79%) primary lung tumors. A combination of CK7 negative, CK20 positive, and TTF-1 and SP-B negative was highly significantly associated with metastatic colorectal carcinomas compared with either primary lung adenocarcinomas or metastatic breast carcinomas (both P < 0.001). A combination of CK7 positive, CK20 negative, and TTF-1 and SP-B negative was highly significantly associated with metastatic breast carcinomas compared with either primary lung adenocarcinomas or metastatic colorectal carcinomas (both P < 0.001).. Use of a panel of antibodies including TTF-1, SP-B, CK7 and CK20 is helpful in discriminating between primary and metastatic adenocarcinomas of the lung and suggesting the primary sites of some metastatic adenocarcinomas.

Topics: Adenocarcinoma; Diagnosis, Differential; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Nuclear Proteins; Pulmonary Surfactant-Associated Protein B; Thyroid Nuclear Factor 1; Transcription Factors

The current authors previously identified circulating cells expressing carcinoembryonic antigen (CEA) messenger ribonucleic acid (mRNA) in 80% of lung cancer patients bearing distant metastases. The current study prospectively validated the data on a novel cohort and extended the study to other mRNAs expressed by neoplastic cells. CEA, cytokeratin 19 and 20, aldolase A and epithelial glycoprotein 2 (EPG2) mRNA was analysed by reverse transcriptase-polymerase chain reaction in circulating cells from 19 healthy controls, and in biopsies and blood at diagnosis from 32 lung cancer patients monitored for 24 months. Aldolase A and cytokeratin 19 mRNA occurred in circulating cells of all controls; cytokeratin 20 was not expressed by any lung cancer biopsy. EPG2 mRNA occurred in all biopsies but not in the patients' circulating cells. CEA mRNA occurred in 29/32 (90.6%) biopsies and in 17/32 mRNA samples from circulating cells from lung cancer patients. Of these positive patients 12/17 developed metastases within 9 months of mRNA analysis. Three positive patients died, one was lost to follow-up, and one did not develop metastases within 24 months. Of the negative patients 12/15 did not develop metastases during the 24-month follow-up; one patient was lost to follow-up, one did not express CEA, and another developed metastases. Unlike in other neoplasias, cytokeratin 19 and 20, aldolase A and epithelial glycoprotein 2 messenger ribonucleic acid are not useful for the detection of circulating cancer cells in lung cancer. Carcinoembryonic antigen messenger ribonucleic acid analysis in circulating cells helps to identify lung cancer patients at a greater risk of metastases.

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Case-Control Studies; Cell Adhesion Molecules; Epithelial Cell Adhesion Molecule; Female; Fructose-Bisphosphate Aldolase; Humans; Intermediate Filament Proteins; Keratin-20; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Neoplasm Staging; Prospective Studies; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Cytokeratin 8 (CK8) is one of the cytoskeletal components and shows caspase-mediated degradation when cells undergo apoptosis. We previously reported that CK8 is highly expressed in non-small cell lung cancer (NSCLC) cell lines and increasing values of serum CK8 are significantly associated with tumor progression in patients with NSCLC. In this investigation, reverse transcriptase-polymerase chain reaction (RT-PCR) analysis in lung cancer cell lines, revealed a shorter PCR product, which differed from the wild-type product of CK8. The nucleotide sequence of the shorter PCR products and genomic DNA for CK8 demonstrated that the shorter product was an aberrantly spliced form of CK8 (AS-CK8) which lacked a caspases cleavage site within the linker lesion in exon 5. The putative protein products predicted by the mRNA of AS-CK8 were demonstrated by Western blotting with monoclonal antibodies for CK8. In addition, AS-CK8 mRNA and its protein products were highly expressed in NSCLC cell lines compared with small-cell lung cancer (SCLC) cell lines. Tissue samples obtained from NSCLC patients also expressed mRNA of AS-CK8. In conclusion, we identified aberrantly spliced CK8 (AS-CK8) which lacked a caspases cleavage site in lung cancer cell lines and primary tumors of NSCLC. AS-CK8 was preferentially expressed in NSCLC, rather than SCLC. These findings lead to speculation that cancer cells expressing AS-CK8 may have a resistance to apoptosis and may perturb keratin network formation.

Topics: Apoptosis; Base Sequence; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Humans; Keratins; Lung Neoplasms; Molecular Sequence Data; Reverse Transcriptase Polymerase Chain Reaction; RNA Splice Sites; RNA, Messenger; Tumor Cells, Cultured

Making a preoperative pathologic diagnosis in patients with small lung nodules remains challenging. We have developed a new, noninvasive bronchoscopic microsampling probe to examine biochemical substances in epithelial lining fluid. We used this probe to measure tumor markers in fluid from tissues surrounding lung nodules less than 30 mm in diameter to test its adjunctive diagnostic utility in lung cancer.. In 12 patients, epithelial lining fluid was collected in triplicate or duplicate from tissue within 2 cm of small peripheral lung nodules and from the contralateral lung. The diagnosis of adenocarcinoma was surgically confirmed in all patients. Fifteen patients without lung cancer served as controls. Concentrations of carcinoembryonic antigen, cytokeratin fragment 19, and sialyl SSEA-1 were measured in the fluid.. Carcinoembryonic antigen and cytokeratin fragment 19 concentrations were significantly higher in fluid near the nodules (median, 8.7 and 87.2 ng/mg, respectively) than on the contralateral sides (median, 1.5 and 3.7 ng/mg, respectively) or in fluid collected from the controls (median, 2.0 and 2.8 ng/mg, respectively).. Measurements of carcinoembryonic antigen and cytokeratin fragment 19 collected by our microsampling probe may be a useful diagnostic adjunct in patients with small peripheral lung nodules.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy, Needle; Bronchoscopes; Bronchoscopy; Carcinoembryonic Antigen; Case-Control Studies; Equipment Design; Female; Humans; Immunohistochemistry; Keratins; Lewis X Antigen; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Prognosis; Prospective Studies; Sensitivity and Specificity; Treatment Outcome

The epithelial alteration in interstitial pneumonias is one of the repair processes at the sites of disease activity. Regenerative epithelial cells may participate in remodeling of the lung. To determine the phenotype of regenerative epithelial cells in usual interstitial pneumonia (UIP) and nonspecific interstitial pneumonia (NSIP), the expression of Clara cell 10KD protein (CC10), cytokeratin (CK) 14 and 17, surfactant apoprotein (SP)-A, KL-6/MUC1, transforming growth factor (TGF) beta2 were examined in 25 patients with UIP, 9 patients with NSIP and normal lung tissues from 10 patients with lung cancer. In honeycomb lesions of UIP, non-ciliated columnar cells mainly expressed CC10, cuboidal cells expressed CC10, CK17, CK14 and SP-A in descending order. Fibroblastic foci are covered by CK17, CK14, CC10, and a few SP-A positive flattened or cuboidal cells. Regenerative epithelium in NSIP mainly comprised cuboidal cells expressing SP-A, CC10 and CK17. KL-6 was more remarkably expressed in cuboidal and non-ciliated columnar cells both in UIP and NSIP. Expression of TGFbeta2 was observed in cuboidal and flattened epithelium. In severe fibrotic areas, CC10 expressing cells were more prominent, while SP-A positive cells were more prominent in less fibrotic areas. Regenerative epithelial cells in remodeling area in UIP may be derived from bronchiolar basal cells and Clara cells, while most of those in NSIP may be derived from type II pneumocytes. The different origin of regenerative epithelium may reflect the severity and extent of the injury and the degree of consequent fibrosis in UIP and NSIP.

Topics: Antigens; Antigens, Neoplasm; Apoproteins; Bronchi; Enzyme Inhibitors; Epithelial Cells; Female; Fibroblasts; Fibrosis; Glycoproteins; Humans; Keratins; Lung; Lung Diseases, Interstitial; Lung Neoplasms; Male; Middle Aged; Mucin-1; Mucins; Phenotype; Phospholipases; Proteins; Pulmonary Surfactant-Associated Proteins; Regeneration; Transforming Growth Factor beta; Transforming Growth Factor beta2; Uteroglobin

We describe a case of Aggressive Digital Papillary Adenocarcinoma, a rare skin neoplasm with acral location.. The patient, a 66 year old man, presented with an ulcerated mass of the fifth toe of the left foot, 4.4 cm in size. Histologically the tumour was characterized by a solid-cystic structure, with extensive papillary component, comedo necrosis and focal eccrine differentiation. Immunohistochemistry showed diffuse positivity for cytokeratins AE1/AE3 and 7, and, focal staining for Muscle Specific Actin, Vimentin and EMA. Chest CT scan showed the presence of a single pulmonary node, whose cytological features were consistent with a metastatic disease.. The clinico-pathological features of the present are similar to those previously reported in the literature and confirms the aggressive nature of this neoplasm.

Topics: Actins; Adenocarcinoma, Papillary; Aged; Biomarkers, Tumor; Cell Differentiation; Foot Diseases; Humans; Keratins; Lung Neoplasms; Male; Mucin-1; Neoplasm Proteins; Sweat Gland Neoplasms; Toes; Vimentin

The purpose of this study was to clarify the significance of bivariate cytokeratin and DNA flow cytometry for analysis of the biologic aggressiveness of resectable non-small cell lung cancer.. In 92 patients who underwent curative operations, the DNA ploidy status and S-phase fractions of the cancer cell populations inside the tumors were analyzed by a cytokeratin gating technique with paraffin-embedded specimens and were correlated with the surgical results.. Ninety tumors yielded assessable DNA histograms. DNA diploidy was detected in 25 tumors with a mean S-phase fraction of 14.3% +/- 4.7%, and DNA aneuploidy was detected in 65 tumors with a mean S-phase fraction of 15.1% +/- 7.1%. The 5-year overall and recurrence-free survivals were 73.3% and 70.3%, respectively. Multivariate analysis showed that only TNM staging was a prognostic factor after surgery. There was a negative correlation between the logarithms of S-phase fraction and the disease-free interval for 22 patients with proven recurrence (P =.006). The tumors with high S-phase fractions recurred more rapidly than did those with low S-phase fractions.. In a bivariate analysis of cytokeratin and DNA flow cytometry in resectable non-small cell lung cancer, the S-phase fraction appeared to be correlated with the disease-free interval. However, DNA ploidy and S-phase fraction were not predictive of either recurrence or survival after operation. Thus DNA flow cytometry may be of limited use for the analysis of the biologic aggressiveness of lung cancer.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; Chi-Square Distribution; DNA, Neoplasm; Female; Flow Cytometry; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Ploidies; S Phase; Statistics, Nonparametric

Shedding of neoplastic cells into the circulation is an essential event for the hematogenous metastasis of solid tumors. Recently, several studies reported that a high frequency of cancer cells could be detected in the bloodstream during surgery. The intraoperative detection of hematogenous dissemination of cancer cells was able to identify a subset of patients with malignant diseases at high risk for postoperative metastasis and to predict a poor prognosis. In order to evaluate the association between intraoperative dissemination of cancer cells and postsurgical survival of patients with non-small cell lung cancer (NSCLC), we developed a flow cytometric assay for specific detection of lung cancer cells in the blood. The monocyte-enriched population in the blood was separated by a modified Ficoll-Hypaque density centrifugation and then labeled with a combination of monoclonal antibodies specific for CD45, cytokeratin (CK) and two antigens expressed on lung cancer cells (2F7 and S5A). The assay could detect quantitatively lung cancer cells (defined as CD45(-1) CK(+) 2F7/S5A(+) cells), with the sensitivity limit of one cancer cell in 10(5) normal leukocytes. The specificity for lung cancer was 97%, which was calculated from the results of healthy subjects (20 cases) and patients affected with benign pulmonary diseases (26 cases) or esophageal cancer (14 cases). Blood samples of 31 NSCLC patients were collected from pulmonary vein during open thoracic surgery. Fifteen of them (48.4%) were found to have positive test results. The average cancer cell counts in these cases were 0.306 x 10(6)/l. Patients under 55 years of age had a significantly higher percentage of positive findings than those over 55 years of age (P < 0.05). The positive rate increased over the stages and lymph node status, but the differences were not statistically significant. Moreover, patients with squamous cell carcinoma at later stages (stages III and IV) had an increased frequency of positive test results than those at earlier stages (stages I and II, P < 0.05). In contrast, no such a difference was found in cases with adenocarcinoma. On the basis of 30-months follow-up date, the median survival time and 2-year survival rate for patients with positive and negative findings were 11 vs. 27 months, and 26.7 vs. 62.5%, respectively. There was a statistically significant difference between overall survival curves that favored the patients with negative test results (P = 0.023). Multivariate

Topics: Aged; Antibodies, Monoclonal; Antigens, Neoplasm; Carcinoma, Non-Small-Cell Lung; Female; Flow Cytometry; Follow-Up Studies; Humans; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Neoplastic Cells, Circulating; Postoperative Complications; Prognosis; Survival

It has been reported that cytokeratin 8 (CK8) can be expressed in several cancers and expression of CK8 is correlated with increased invasiveness of the tumor in vitro and in vivo. In the present study, we investigated expressions of CK8 in human lung cancer cell lines. In addition, we also evaluated the clinical significance of CK8 measurements in sera of patients with lung cancer. Expression of mRNA for CK8 was semi-quantitatively evaluated by the competitive reverse transcriptase-polymerase chain reaction (competitive RT-PCR), using human lung cancer cell lines. The level of CK8 protein in culture supernatants of lung cancer cell lines and 70 sera of patients with lung cancer was measured by enzyme-linked immunosorbent assay (ELISA). Levels of serum CK8 according to clinical parameters were also examined. The level of expression of CK8 mRNA in non-small cell lung cancer (NSCLC) cell lines was significantly high compared with that of small cell lung cancer (SCLC) cell lines (P<0.05). The level of CK8 in culture supernatants in NSCLC was significantly high compared with that of SCLC. The level of serum CK8 in patients with NSCLC was significantly high compared with that of normal non-smokers and compared with that of SCLC (P<0.05). Patients with a CK8 value of 50.0 ng/ml, or higher, had a statistically significant diminished survival compared with those patients whose CK8 values were lower. In conclusion, CK8 was preferentially expressed in NSCLC. Increasing values of CK8 were significantly associated with tumor progression and decreased survival in patients with NSCLC. Therefore, CK8 in sera may become a novel tumor marker in patients with lung cancer.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Disease Progression; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression Regulation, Neoplastic; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; Survival; Tumor Cells, Cultured

Mucinous bronchioloalveolar carcinomas (BACs) can closely mimic metastatic adenocarcinoma to the lung both clinically and morphologically. Several studies have demonstrated that the differential expression of cytokeratin 7 (CK7) and cytokeratin 20 (CK20) is a valuable diagnostic tool in differentiating primary pulmonary adenocarcinomas (PPAs) (usually CK7 positive/CK20 negative) from metastatic colonic adenocarcinoma (usually CK7 negative/CK20 positive). The present study is designed to correlate the histologic subtypes of PPA with expression of 7 and 20. A total of 113 cases of bonafide PPA were selected and classified according to the 1999 World Health Organization criteria as adenocarcinoma, NOS (n = 80), nonmucinous BAC (n = 14), and mucinous BAC (n = 19). Representive sections of all the tumors were immunohistochemically analyzed for CK7 and CK20 expression. To evaluate the diagnostic utility of CK7 and CK20 expression, 6 cases of colonic adenocarcinoma metastatic to the lung were tested with the same antibodies and compared with mucinous BAC. Results were expressed in a semiquantitative fashion based on the percentage of positive tumor cells: <10%, focal; 10% to 25%, 1+; 26% to 75%, 2+; > or =76%, 3+. All 113 PPAs exhibited strong, diffuse CK7 expression. With respect to CK20 expression, 17 of the 19 cases (89.4%) of mucinous BAC showed moderate to strong expression of this protein, whereas only 10 cases of conventional adenocarcinomas and 4 cases of nonmucinous BAC exhibited expression. All 6 examples of metastatic colonic adenocarcinomas were negative for CK7 and strongly positive for CK20. In summary, mucinous BAC is distinct from other PPAs by virtue of its CK20 expression. Although the CK7/CK20 immunoprofile is a valuable diagnostic marker for differentiating primary lung adenocarcinoma from metastatic colonic adenocarcinoma, caution should be exercised when dealing with mucinous BAC.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Adenocarcinoma, Mucinous; Biomarkers, Tumor; Colonic Neoplasms; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Retrospective Studies

Carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC-Ag), and CYFRA 21-1 are the most useful markers for non-small cell lung cancer (NSCLC), but neuron-specific enolase (NSE) is a tumor maker of choice for SCLC. The determination of NSE in NSCLC could allow selection of patients with neuroendocrine features. NSCLC patients whose tumors have neuroendocrine properties may be more responsive to chemotherapy; however, these tumors have been reported to be more aggressive. Tumor markers are not suitable for diagnosis; their principal applications are in monitoring of therapy and prognosis.. Tumor markers were measured in 200 untreated patients with squamous cell lung cancer (SQC) and a reference group (n = 220; 124 healthy persons and 96 patients with nonmalignant lung disease). CEA and SCC-Ag were measured by microparticle enzyme immunoassays on Abbott AxSYM and IMx analyzers. CYFRA 21-1 and NSE were measured by electrochemiluminescence immunoassays on the Roche Elecsys 2010.. CEA, SCC-Ag, CYFRA 21-1, and NSE were increased above the cutoffs in 26%, 32%, 67%, and 28% of tested patients, respectively. The area under the ROC curve for CYFRA 21-1 was higher than those for CEA, SCC-Ag, and NSE (SQC vs controls). CYFRA 21-1 and CEA were significantly higher in advanced SQC than in early stages of disease (P <0.0001 and P <0.0004, respectively). In multivariate analysis of survival, CYFRA 21-1 was an independent but nonspecific prognostic factor in the operable group of SQC patients, whereas NSE was an independent prognostic factor in the advanced stages of disease.. CYFRA 21-1 is an independent prognostic factor in earlier stages and NSE in the advanced stages of SQC.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Neoplasm Staging; Phosphopyruvate Hydratase; ROC Curve; Sensitivity and Specificity; Serpins; Survival Rate

Primary squamous cell carcinoma of the endometrium (PSCCE) is an exceedingly rare tumor. Rarely are cytological criteria discussed. We report our experience in the cytological diagnosis of a case. A postmenopausal, 64-yr-old woman suffered from pyometria. An endometrial Pap smear displayed some malignant squamous cells. Curettage of the cervix and the uterine cavity only recovered some fragments of atypical squamous epithelium whose origin could not be precisely identified. A hysterectomy with bilateral adnexectomy was decided upon. Pathological study evidenced a primary squamous cell carcinoma in the uterine cavity while the cervix was tumor-free and the lymph nodes were devoid of metastases (pT1, pN0, pM0). The patient died 46 mo PO with multiple pulmonary and renal metastases. The histological feature of PSCCE is identical to that of any tumor of a similar nature, whatever the site, especially the cervix. Confirmation of the primary endometrial nature is only possible on the hysterectomy specimen.

Topics: Carcinoma, Squamous Cell; Dilatation and Curettage; Endometrial Neoplasms; Fatal Outcome; Female; Humans; Hysterectomy; Immunohistochemistry; Keratins; Kidney Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Middle Aged; Mucin-1; Papanicolaou Test; Vaginal Smears

Cellular keratin, an indicator for cancer cell in circulation gains increasing attention. Although CK19mRNA as a gene marker used to identify micro-metastasis is of relatively high practical value, it lacks specificity and sensitivity in diagnosis of cancer. The aim of this study is to evaluate whether the combined detection of CK19mRNA with other tumor markers, such as carcinoembryanic antigen(CEA), neuron-specific Enolase(NSE), tissue polypeptide antigen(TPA), would improve the diagnosis of lung cancer.. The expression level of CK19mRNA in peripheral blood mononucleated cells (PBMCS) was determined by RT-PCR, The levels of CEA, NSE, and TPA in serum were determined by enzyme-linked immunosorbent assay (ELISA) and time-resolved fluorometry(TRF) in 91 patients with lung cancer, 33 with benign lung diseases (BLD) and 50 healthy controls. The data was analyzed by t test and chi 2 test.. The positive rates of CK19mRNA, CEA, NSE, and TPA in lung cancer group (49.5%, 65.9%, 67.0%, and 83.5%, respectively) were remarkably higher than in BLD group(24.2%, 9.0%, 15.2%, and 9.0%, respectively) and in healthy control group (10.0%, 4.0%, 8.0%, and 6.0%, respectively) (P < 0.01). The positive rate of CK19mRNA was not statistically different in various histological subtypes (P > 0.05). Although the positive rate of CK19mRNA appeared to be associated with the clinical stage (stage I + II: 44.4%, stage III: 48.4%, stag IV: 54.2%), there was no statistical significance among the three groups (P > 0.05). The serum level (microgram/L)/positive rate of CEA(3.5/27.8%, 5.1/35.5%, and 8.5/58.3%, respectively), NSE (12.5/33.3%, 15.3/41.9%, and 24.7/58.3%, respectively), and TPA (1.1/44.4%, 1.8/58.1%, and 3.6/66.7%, respectively) in stage I + II, III, and IV appeared to be associated with the clinical stages of lung cancer, but no statistical significance among the three groups (P > 0.05). Combined detection of three or four tumor markers yielded significant higher effective accuracy in diagnosis of lung cancer than that of any of two tumor markers or single tumor marker (P < 0.05). In addition, the results showed that TPA, CEA, and NSE were relatively specific for squamous cell carcinoma (76.7%), adenocarcinomas (83.3%), and small-cell lung cancer (79.2%), respectively.. The combined measurement of CK19mRNA with CEA, NSE, and TPA can increase diagnosis rate of lung cancer, can also provide potent diagnosis basis for treatment.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoembryonic Antigen; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; RNA, Messenger; Tissue Polypeptide Antigen

The purpose of this study was to evaluate the markers' clinical usefulness for early prediction of recurrence, by serial and simultaneous measurements of serum cytokeratin fragment 19 (CYFRA 21-1), before and after surgery on patients with non-small cell lung cancers (NSCLC). The 48 patients enrolled in this study had adenocarcinoma of the lung (adenoCa) (including 24 patients with recurrence and 24 patients without recurrence 1 year after surgery) and 48 patients with squamous cell carcinoma of the lung (SCC) (including 24 cases with recurrence and 24 without recurrence 1 year after surgery). Serial serum levels of CYFRA 21-1 were measured before the operation and 1 week, 1 month, 3 months, 6 months, 9 months, and 12 months after surgery for the early detection of recurrence. The results revealed that (1) the mean serum values of CYFRA 21-1 were significantly higher beginning at 1 month after surgery in the 24 patients with recurrent adenoCa compared with the 24 patients without recurrent adenoCa, (2) mean serum values of CYFRA 21-1 were significantly higher beginning at 1 month after surgery in 24 patients with recurrent SCC when compared with 24 patients without recurrent SCC, and (3) mean serum values of CYFRA 21-1 were significantly higher beginning at 1 month after operation in the total 48 patients with recurrent NSCLC when compared with 48 patients without recurrent NSCLC. We conclude that CYFRA 21-1 is not a good marker for early prediction of NSCLC recurrence including adenoCa and SCC after surgery.

Topics: Adenocarcinoma; Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Predictive Value of Tests; Retrospective Studies

Urothelial carcinoma with choriocarcinomatous features is a rare malignancy arising in the urinary bladder or renal pelvis. We report the case of a 60-year-old man with a biphasic neoplasm of the right kidney composed of papillary urothelial carcinoma and choriocarcinoma. Widespread hepatic and pulmonary metastases with choriocarcinomatous features were found on autopsy 6 weeks after initial diagnosis. Chromosomal analysis revealed a close genetic relationship between the papillary urothelial and choriocarcinomatous tumor components, documenting for the first time that choriocarcinoma of the renal pelvis results from clonal evolution of urothelial carcinoma with acquisition of trophoblastic differentiation.

Topics: Carcinoma, Transitional Cell; Choriocarcinoma; Chromosome Aberrations; Chromosomes, Human, Pair 12; Chromosomes, Human, Pair 17; Chromosomes, Human, Pair 4; Chromosomes, Human, Pair 9; Gene Amplification; Gene Deletion; Histocytochemistry; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Nucleic Acid Hybridization

In 27 patients with operable non-small cell lung cancer (NSCLC) submitted to radical surgery followed by 3 cycles of chemotherapy (cht) serum concentrations of Cyfra 21.1 and TPA were studied. The measurements were performed before and 14 days after surgery, before each cht and every 60th day after cht was completed, for 2 years. Seven patients died during the follow up. There was no significant correlation between preoperative cyfra 21.1 and TPA serum concentrations and stage of diseases or histologic types of NSCLC. Initial concentrations of the two markers had no prognostic meaning. A significant decrease of 2 markers was observed after surgery in the whole group and in patients with therapy success. While adjuvant cht did not influence significantly serum concentrations of the markers, we showed a significant elevation of 2 markers about 4 months before death. It seems that establishing of values of Cyfra 21.1 and TPA in the patient's follow up may be useful in recognition of tumour relapse.

Topics: Adult; Aged; Antigens, Neoplasm; Antineoplastic Combined Chemotherapy Protocols; Antineoplastic Protocols; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Chemotherapy, Adjuvant; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Quality of Life; Time Factors; Tissue Polypeptide Antigen

To detect occult micrometastatic tumor cells in pN0 lymph nodes of nonsmall cell lung cancer (NSCLC) by a combination of cytokeratin and p53 immunohistochemistry staining, and to evaluate the relation between the micrometastasis in pN0 lymph nodes and the prognosis of patients with completely resected stage 1 NSCLC.. The average 5-year survival rate for patients with completely resected stage 1 NSCLC is only about 70%; thus, about 30% of these patients have recurrent disease. This suggests that occult micrometastasis may exist at the time of surgery; the rate is clearly underestimated by current clinical staging examinations and conventional histopathologic methods.. A total of 474 hilar and mediastinal lymph nodes were removed during surgery from 49 patients with completely resected stage 1 NSCLC. The lymph nodes analyzed for micrometastasis using immunohistochemical staining with the biclonal anticytokeratin antibody, AE1/AE3. Of these 474 lymph nodes from 49 patients, 263 lymph nodes from 25 patients, whose primary tumors were positive for the p53 protein, were subjected to immunohistochemical staining with the monoclonal anti-p53 protein antibody DO-1.. Cells positive for cytokeratin and p53 protein were found in 35 (7.4%) of 474 and 20 (7.6%) of 263 lymph nodes, respectively; 17 (34.7%) of 49 patients had cytokeratin-positive cells and 10 (40.0%) of 25 patients had p53-positive cells in their pN0 lymph nodes. By a combination of cytokeratin and p53 protein immunohistochemical staining, micrometastatic tumor cells were identified in pN0 lymph nodes in 22 (44.9%) of 49 patients. The patients with lymph node micrometastasis identified by a combination of cytokeratin and p53 protein immunohistochemical staining had a poorer prognosis than those without micrometastasis on both univariate and multivariate analyses (overall survival, P =.0003 and 0.013, respectively).. The detection of lymph nodal micrometastasis by cytokeratin and p53 protein immunohistochemical staining will be helpful to predict the recurrence and prognosis of patients with completely resected stage 1 NSCLC.

Topics: Carcinoma, Non-Small-Cell Lung; Chi-Square Distribution; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Recurrence, Local; Prognosis; Proportional Hazards Models; Regression Analysis; Survival Analysis; Time Factors; Tumor Suppressor Protein p53

Systemically disseminated tumor cells have become the subject of intensive research as the presumed seminal precursors of later distant metastasis. We describe here a novel sensitive multimarker nested reverse transcription (RT)-PCR capable of detecting the individual expression of human MAGE-A genes MAGE-1, -2, -3/6, -4, and -12 by rare, disseminated tumor cells in bone marrow and blood of patients with many different types of cancer. We analyzed bone marrow aspirates from 106 patients with breast, lung, colorectal, and prostate cancer and with different sarcomas. Heterogeneous expression of the different MAGE genes was found frequently in all those kinds of malignancies, in sharp contrast to 30 bone marrow and 20 blood samples from healthy donors, which were completely MAGE negative. Expression of at least one MAGE gene in bone marrow was more frequent than cytokeratin-positive tumor cells detected by immunocytochemistry, although the results of both tests overlapped considerably. In 30 patients with clinically localized prostate cancer, analysis by the multimarker MAGE RT-PCR of bilateral bone marrow aspirates from the right and left iliac crest revealed a positivity rate of 60%, which was twice as high as that obtained with either an established prostate-specific antigen RT-PCR or by cytokeratin-based immunocytochemistry. Analysis of primary prostate cancer revealed MAGE expression patterns considerably concordant with those found in the corresponding bone marrow aspirates. Prostate cancer patients carrying an exceptionally high risk of metastatic relapse, as defined by clinical prognostic factors, were significantly more often MAGE positive than patients with a distinctly lower risk (P = 0.02, Fisher's exact test). More frequent MAGE expression in the peripheral blood of patients with metastatic prostate cancer compared with those with clinically localized disease added further evidence for the prognostic impact of the multimarker MAGE RT-PCR. Moreover, MAGE-positive bone marrow samples from a small group of seven sarcoma patients demonstrated the relevance of our multimarker RT-PCR in nonepithelial tumors. Because MAGE antigens can induce autologous cytolytic T lymphocytes in vivo, the determination of individual MAGE expression patterns in cancer patients may furthermore identify candidate vaccine targets for adjuvant immunotherapy.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Breast Neoplasms; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Neoplasm Metastasis; Prostate-Specific Antigen; Prostatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; RNA, Messenger

The presence of disseminated tumor cells in bone marrow is considered to be a premetastatic state, which is called micrometastasis. To evaluate the relationship between micrometastasis and cellular adhesion molecules in the primary lesion, E-cadherin and beta-catenin were immunohistochemically investigated. Methods. Fifty-eight patients with non-small cell lung cancer who underwent a complete resection were entered into this study. Tumor cells in bone marrow aspirates were detected by immunohistochemistry using cytokeratin (CK) 18. Immunohistochemical studies of E-cadherin and beta-catenin were performed in the corresponding primary tumor.. CK-positive cells were detected in the bone marrow aspirates from 27 of 58 patients. A reduced expression of the E-cadherin and beta-catenin was found in 16 (27.6%) and in 22 (37.9%) of 58 patients, respectively. In 26 cases with a reduced expression of E-cadherin and/or beta-catenin, 16 cases had CK-positive cells, whereas 11 of 32 cases with normal expression of both factors had CK-positive cells (P=.0392). The patients with micrometastasis demonstrated an earlier recurrence (P =.0642) and a significantly poorer survival (P =.0437) than those without such cells.. Micrometastasis in the bone marrow might be a significant predictor of poor prognosis, and a reduced expression of E-cadherin and beta-catenin are important determinants for the metastatic capability of individual cancer cells.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; beta Catenin; Bone Marrow Neoplasms; Cadherins; Carcinoma, Squamous Cell; Cytoskeletal Proteins; Disease-Free Survival; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Prognosis; Risk Factors; Survival Rate; Trans-Activators

The role of tumor markers in the diagnosis and prognosis of lung cancer is under investigation.. The aim of this study was to investigate the diagnostic and prognostic significance of pre-therapeutic levels of various serum tumor markers, CYFRA 21-1, neuron-specific enolase (NSE), tissue polypeptide antigen (TPA), carcinoembryonic antigen (CEA), CA 125 and squamous cell carcinoma antigen (SCCAg), in patients with lung cancer.. We studied 102 consecutive patients (mean age 65.2 +/- 11 years) with newly diagnosed lung cancer (96 males, 94%, with a mean age of 66.3 +/-10.5 years). All patients had a 5-year follow-up. Measurements of the serum tumor markers were performed on initial diagnosis.. Eighty-four patients (82%) had non-small-cell lung cancer (NSCLC) and 18 (18%) small-cell lung cancer (SCLC). From the 84 patients with NSCLC, 34 patients (33%) had squamous-cell lung cancer, 23 (22%) adenocarcinoma and 23 (22%) large-cell carcinomas. The overall median survival was 8.5 months. All SCLC patients had extensive disease with a median survival of 10.1 months and NSCLC patients of 8.4 months. Significant differences in the mean values of NSE and CYFRA 21-1 were observed between SCLC and NSCLC. In NSCLC, CYFRA 21-1, TPA, CA 125 and SCCAg serum levels were related to the stage of the disease at diagnosis, and CYFRA 21-1, NSE, TPA and CA-125 were related to a poor outcome. None of the above tumor markers was related to survival in the SCLC group.. CYFRA 21-1 and NSE may help to differentiate cell types in lung cancer patients. Also, CYFRA 21-1 with TPA and CA 125 may provide useful information regarding the staging of the disease at diagnosis and the prognosis of patients with NSCLC.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Analysis of Variance; Antigens, Neoplasm; Biomarkers, Tumor; CA-125 Antigen; Carcinoembryonic Antigen; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Probability; Prognosis; Prospective Studies; Regression Analysis; Sensitivity and Specificity; Survival Analysis

Micropapillary carcinoma or a micropapillary carcinoma component has been reported in the ovary, breast, and urinary bladder and is generally thought to have prognostic significance. However, little has been written on micropapillary differentiation in lung carcinoma. We studied 35 cases of primary lung adenocarcinoma with a micropapillary component seen at the M.D. Anderson Cancer Center. The micropapillary component in these tumors ranged from focal to prominent and was seen at both primary and metastatic sites. This component was not associated with any particular histologic subtype of lung adenocarcinoma. Of the 15 cases with available material, 14 (93%) stained positive for cytokeratin 7, whereas only two of the 15 cases (13%) stained positive for cytokeratin 20. Thyroid transcription factor-1 immunostaining of tumor nuclei was seen in 12 of the 15 cases (80%). Immunostaining was seen in areas both with and without micropapillary differentiation. Thirty-three of 35 patients (94%) developed metastases, which occurred most commonly in the lymph nodes (n = 26), and also in the lung (n = 17), brain (n = 9 cases), bone (n = 9 cases), and other sites. Most metastases had a prominent micropapillary component, irrespective of the extent of the micropapillary carcinoma component in the primary lung tumor. Adequate clinical follow-up information was available for 29 patients. The mean follow-up was 25 months. At their last follow-up, 16 of 29 patients (55%) were still alive with disease, 5 (17%) were dead of disease, and 8 (28%) were alive with no evidence of disease. We believe that a micropapillary component occurring in lung adenocarcinoma should be reported, as this component may be more likely to metastasize. The presence of this component should alert the clinician to search more carefully for metastases and have a closer follow-up on these patients. It is also important to recognize this component in evaluating a metastasis from an unknown primary site, as it should alert the pathologist to a possible primary in the lung in addition to breast, urinary bladder, and ovary.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Bone and Bones; Carcinoma, Papillary; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Male; Middle Aged; Neoplasm Metastasis; Nuclear Proteins; Prognosis; Thyroid Nuclear Factor 1; Transcription Factors

There are many reports of sclerosing hemangioma from the perspective of its histopathologic features, but its cytopathologic characteristics are less well known. In this report we present the case of a patient in which the cytologic features firmly established a definitive diagnosis; surgical intervention was warranted only after the lesion had grown over the course of 7 yr of close observation. The cytologic diagnosis requires the identification of a dual cell population. Both populations of tumor cell nuclei are immunoreactive for thyroid transcription factor-1, but caution is warranted because this marker may be present in other tumors. Recognition of its distinctive cytologic features can lead to proper diagnosis and conservative management.

Topics: Biomarkers, Tumor; Biopsy, Needle; Female; Histiocytoma, Benign Fibrous; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Neoplasm Proteins; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors; Treatment Outcome; Vimentin

Pulmonary metastases of endometrial stromal sarcoma (ESS) are uncommon and can pose diagnostic problems. We reviewed lung specimens from 16 patients with metastatic ESS. Patients were 31-77 years of age at the time of lung biopsy. Uterine ESSs were diagnosed an average of 9.8 years before lung biopsy in 11 patients. Uterine ESSs were originally called smooth muscle tumors in three additional patients. Thirteen patients were evaluated for new pulmonary nodules, seven of whom were asymptomatic. Nodules were multiple in 14 and solitary in four, ranging from 1.0 to 8.0 cm in greatest dimension. One patient died of metastatic disease; 14 were alive and seven of these were without disease (mean follow-up 4.1 years). Diagnostic considerations in 12 consultation cases included ESS, sclerosing hemangioma, carcinoid tumor, lymphangioleiomyomatosis, endometriosis, hemangiopericytoma, and lymphoma. Tumors were well circumscribed and usually solid, composed of plump spindle cells arranged in short fascicles. Two tumors were predominantly cystic. Sex cord-like stromal differentiation was identified in three. Neoplastic cells stained for vimentin (93%), estrogen and progesterone receptor (100%), smooth muscle actin (57%), desmin (50%), and keratin (46%). Metastatic ESS should be included in the differential diagnosis of nonepithelial neoplasms in women.

Topics: Actins; Adult; Aged; Desmin; Endometrial Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Receptors, Estrogen; Receptors, Progesterone; Sarcoma, Endometrial Stromal; Vimentin

Due to more efficient chemotherapy protocols, the number of second and even third primary carcinomas is steadily increasing. To denominate the possible origin of a carcinoma, different markers are available as an aid, e.g. hormones, proteins and lipoproteins, secretion products and cytoskeletal proteins. Cytokeratins (CKs) have gained new popularity; however, they have not been extensively evaluated in lung tumours. In our study we evaluated the staining patterns of CK polypeptides 4-8, 10, 13, 14, and 17-20 and high molecular weight (HMW) CK polypeptides in routinely processed primary lung carcinomas and lung metastases of diverse origin. As expected, immunohistochemical investigation gave no clear-cut results, but, with statistical analysis, lung adenocarcinomas could be separated from metastatic adenocarcinomas using CK 5 and 18 and HMW CK (specificity 92.5%, sensitivity 62.5%). The different origin of the metastases could often be detected using CK 18 and CK 20. Lung clear cell carcinomas and large cell carcinomas with clear cell areas could be distinguished from metastatic renal clear cell carcinomas by the CK 7 staining reaction. Squamous cell carcinomas of the lung and metastatic squamous cell carcinomas of the larynx, pharynx and oesophagus could not reliably be separated in part due to the few number of cases available. CK polypeptide typing is thus an additional aid in the differential diagnosis of lung carcinomas versus carcinomas metastatic to the lung.

Topics: Adenocarcinoma; Carcinoma, Squamous Cell; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms

Adenocarcinoma (AC), squamous cell carcinoma (SCC) and adenosquamous carcinoma (ASC) of the lung are morphologically distinguished in part by cyto-architectural features. However, little is known about the relative expression and distribution of cyto-architectural proteins among AC, SCC and ASC. Initial microarray analysis revealed significant differences in expression of two cyto-architectural genes in AC, SCC and ASC. Desmoplakin (DP) 1 and 2, which link desmosomes to intermediate filaments, was strongly expressed in SCC relative to AC and ASC. Cytokeratin 18 (CK18), an intermediate filament that is commonly linked to desmoplakin, was strongly expressed in AC and ASC relative to SCC. Western blot analysis demonstrated that AC and ASC had abundant CK18 protein, whereas CK18 was weakly detected in SCC. DP 1 and 2 are strongly expressed in SCC and minimally expressed in AC and ASC. However, the ratio of one to the other is the same in SCC and AC, but DP2 is lost in ASC. Microscopic analysis with fluorescence-labeled antibodies for CK18 and DP 1 and 2 revealed abundant membrane localization of DP and minimal perinuclear localization of CK18 in SCC. In contrast, in both AC and ASC, the CK18 protein was diffusely distributed within the cytoplasm, and DP showed both membranous and cytoplasmic localization. In conclusion, the data here shows that AC, SCC and ASC each have specific patterns of DP 1 and 2 and CK18 gene expression, protein content and biodistribution.

Topics: Biomarkers, Tumor; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Cytoskeletal Proteins; Desmoplakins; Gene Expression Profiling; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Microscopy, Confocal; Microscopy, Fluorescence; Oligonucleotide Array Sequence Analysis; Prognosis; Tissue Distribution; Tumor Cells, Cultured

Carcinoembryonic antigen (CEA), cancer antigen 125 (CA 125), NCC-ST-439, carbohydrate antigen 19-9 (CA 19-9), cytokeratin 19 fragment (CYFRA 21-1), sialyl Lewis X-i antigen (SLX), progastrin-releasing peptide (ProGRP), squamous cell carcinoma antigen (SCC) and neuron specific enolase (NSE) were evaluated in the pleural effusion of 39 patients with lung cancer (29 adenocarcinomas, seven small-cell carcinomas, three squamous cell carcinomas) and 43 patients with tuberculous pleurisy. The levels of the tumor markers other than SCC and NSE were significantly higher in lung cancer than in tuberculosis. High levels of CYFRA 21-1 and SCC were observed in squamous cell carcinoma and high levels of ProGRP and NSE were observed in small-cell carcinoma. According to the validity score, sensitivity (%) + specificity (%) - 100, the optimal cut-off levels of pleural effusion were 8.1 ng/ml for CEA, 660 U/ml for CA 125, 2.6 U/ml for NCC-ST-439, 10 U/ml for CA 19-9, 65 ng/ml for CYFRA 21-1, 140 U/ml for SLX, 23.2 pg/ml for ProGRP, 0.6 ng/ml for SCC and 5 ng/ml for NSE. By comparison of validity scores for each optimal cut-off level and of receiver operating characteristic (ROC) curves, we suggest that a CEA assay is the most useful for pleural effusion. The combined assay of CEA + ProGRP and CEA + ProGRP + CYFRA 21-1 were considered to be useful.

Topics: Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; CA-125 Antigen; CA-19-9 Antigen; Carcinoembryonic Antigen; Humans; Keratin-19; Keratins; Lung Neoplasms; Oligosaccharides; Peptide Fragments; Peptides; Phosphopyruvate Hydratase; Pleural Effusion, Malignant; Recombinant Proteins; Serpins; Sialyl Lewis X Antigen; Tuberculosis, Pleural

To study the influence of micrometastasis in lymph node on staging and prognosis of non-small-cell lung cancer (NSCLC).. In 39 NSCLC patients, micrometastasis in pathologically negative lymph nodes were tested through immunohistochemical cytokeratin (CK) analysis and the relationship between CK(+) and staging, survival were analyzed.. In these 39 patients, the survival of CK(+) and CK(-) patients were 32 months and 48 months respectively (P = 0.0178). Multivariate analysis of Cox regression model showed: clinical stage (P = 0.0288) and relapse or metastasis (P = 0.0053) affected the prognosis while micrometastasis in lymphnodes (P = 0.7740) did not.. The detection of micrometastasis in the lymphnodes may serve as a supplement to the present staging system for lung cancer. Even though the prognosis of patients with micrometastasis being poorer than those without, micrometastasis in the lymph nodes should not be regarded as an independent prognostic factor.

Topics: Adult; Aged; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratins; Lung Neoplasms; Lymph Nodes; Male; Middle Aged; Neoplasm Staging; Prognosis

The aim of this prospective study was to assess the prognostic impact of serum tumor markers (Cyfra21-1, carcinoembryonic antigen, neuron-specific enolase, squamous cell carcinoma-antigen and TPAcyk) in patients with non-small cell lung cancer (NSCLC) receiving complete resection.. Sixty-seven patients with histologically proven NSCLC and complete resection of stage I-IIIA disease were included. The serum levels of all markers were measured using commercially available immunoassays.. With a median follow-up of 86 months for surviving patients, those with initial Cyfra21-1 serum levels higher than 3.57 ng/ml had a significantly worse prognosis (P=0.014). The remaining serum tumor markers showed no prognostic impact. In a Cox regression model, Cyfra21-1 proved to be an independent prognostic factor for both overall survival and disease-free interval. In addition, Cyfra21-1 sustained as an independent prognostic factor in completely resected stage I/II disease.. With a cut-off value of 3.57 ng/ml, Cyfra 21-1 was an independent prognostic factor for survival in NSCLC-patients with complete resection. Further evaluation is needed, particularly in stage I/II disease. When the prognostic impact is confirmed with larger patient numbers this may contribute to the identification of stratification variables for future treatment approaches of NSCLC.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Follow-Up Studies; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Phosphopyruvate Hydratase; Prognosis; Proportional Hazards Models; Prospective Studies; Regression Analysis; Serpins; Survival Analysis; Tissue Polypeptide Antigen

To evaluate the effect of vascular endothelia1 growth factor (VEGF) on the hematogenous metastasis of non-small cell lung cancer (NSCLC).. The identification of lung cancer cells in the peripheral blood were carried out by cytological, immunohistocytologica1 and immunofluorecent stains respectively, following isolation of cytokeratin-expressing cells with magnetic activated cell sorting. The quantification of cancer cells in the blood was performed according to the established flow cytometric assay. The plasma VEGF was measured by commercially available ELISA kit.. The lung cancer cells in the blood, showing a remarkable nuclear polymorphism, expressed the epithelial marker cytokeratin and telomerase reverse transcriptase (hTERT). These cells were stained positive by an NSCLC-specific monoclonal antibody S5Al0-2, but negative by antibodies against CD34 and CD45 antigens. Using the flow cytometric assay, 44 cases (28.6%) of l54 NSCLC patients were found to have cancer cells in their blood, with the incidence of positive cases correlated with the stage of disease. The plasma VEGF level was significantly increased in NSCLC patients in comparison with healthy individuals and patients with benign pulmonary diseases. This level was correlated with the stages of disease in patients with adenocarcinoma. In patiens with cancer cells in their blood, a higher level of plasma VEGF was related with an increased number of cancer cells.. The plasma VEGF level is increased in NSCLC patients with approximate1y one fourth to have cancer cells in the peripheral blood. In these patients, increased VEGF level promotes hematogenous tumor metastasis, as indicated by a much higher number of cancer cells in the blood.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; DNA-Binding Proteins; Endothelial Growth Factors; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphokines; Male; Middle Aged; Neoplasm Metastasis; Neoplastic Cells, Circulating; Telomerase; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

In surgically resected specimens of squamous cell carcinoma (SCC) of the lung from 45 patients, we immunohistochemically examined the expression of 13 subtypes of cytokeratin (CK), the intermediate filament in cytoplasm of epithelial cells. To investigate heterogeneity of CK, its expression was compared among tumor cell nests with or without keratinization and stratification. Furthermore, the relationship between CK expression and Ki-67 labeling index or p53 expression was investigated. The tumor cell nests with keratinization showed the expression of CK1 and CK10 as a central pattern and the expression of CK14 as a peripheral pattern. The nests with stratification showed CK14 expression as a peripheral pattern, whereas those without stratification showed the expression as a diffuse pattern. The tumor cell nests showing stromal invasion with fibrosis in the marginal zone were diffusely positive for CK14. Ki-67 antigen labeling index was significantly higher in the nests where CK14 expression was diffuse or peripheral than in the nests where the expression was focal or negative. In lymph node metastases, the tumor cells often showed CK14 expression, like trabecular nests in the primary carcinoma. These results suggest that CK14 is a parameter of proliferative activity and metastatic potential of SCC of the lung.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Division; Female; Humans; Immunoenzyme Techniques; Keratin-14; Keratins; Ki-67 Antigen; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Tumor Suppressor Protein p53

The origin of metastatic adenocarcinoma lesions can sometimes be difficult to diagnose. The objectives of our study were to establish the cytokeratin staining pattern of primary and metastatic lung and colorectal adenocarcinomas, and to determine if this helps to identify the site of origin of metastatic lesions. We reviewed a total of 102 tissue samples from patients in our tumour registry, with either primary or metastatic lung or colorectal adenocarcinoma. Tissue sections were stained for cytokeratin 7 and 20 and read as positive or negative for staining. Clinical and radiologic information was reviewed from computerised charts. The cytokeratin 7+/cytokeratin 20- pattern characterised 96% (29 out of 30) of primary and 95% (21 out of 22) of metastatic lung adenocarcinomas. All the primary (26), and 88% (21 out of 24) of metastatic colorectal adenocarcinomas stained cytokeratin 7-/cytokeratin 20+. Samples from a variety of metastatic sites were evaluated for cytokeratin 7 and 20 staining. Out of the 102 samples, in 95% (97 out of 102) of the cases, the cytokeratin 7 and cytokeratin 20 staining pattern characterised and differentiated between lung and colorectal adenocarcinoma. Primary and metastatic lung adenocarcinomas show a cytokeratin 7+/cytokeratin 20- staining pattern, while colorectal adenocarcinomas stain cytokeratin 7-/cytokeratin 20+. Cytokeratin staining is helpful in the diagnostic differentiation of metastatic lesions from these two common primaries, and assists in determining the site of origin of metastatic lesions.

Topics: Adenocarcinoma; Biopsy; Colorectal Neoplasms; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Prognosis; Registries; Staining and Labeling

Determining the site of origin of metastatic adenocarcinoma lesions has obvious therapeutic and prognostic implications. Immunostaining for cytokeratin subtypes 7 and 20 is increasingly used to evaluate metastatic colorectal cancer lesions. We present the case of a patient with a history of colorectal cancer who subsequently developed an isolated metastatic lesion in the lung. As illustrated by our case, staining for cytokeratin subtypes can be helpful in determining the site of origin of metastatic lesions.

Topics: Adenocarcinoma; Aged; Cecal Neoplasms; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Neoplasms, Unknown Primary

Immunohistochemistry provides an important indicator for differential diagnosis between pleural malignant mesothelioma and lung adenocarcinoma, which have complex therapeutic and medicolegal implications. To pinpoint a reliable, restricted panel of markers, the authors evaluated the efficacy of select commercial antibodies in a series of patients with confirmed clinicopathologic diagnosis of mesothelioma or lung adenocarcinoma with the aid of multiple logistic classification tables. Specimens of 46 mesotheliomas and 20 lung adenocarcinomas were examined with calretinin, thrombomodulin, cytokeratins (CKs) 5/6, and high-molecular weight CKs (indicators of mesothelioma), alongside MOC 31, Ber-EP4, and carcinoembryonic antigen (CEA; indicators of lung adenocarcinoma). Of the mesotheliomas, 40 of 46 (87%) were positive with calretinin, 29 of 46 (63%) with thrombomodulin, 40 of 46 (87%) with CKs 5/6, and 41 of 46 (89%) with high-weight CKs; five of 46 mesotheliomas (11%) were focally reactive with MOC 31, four of 46 (9%) with Ber-EP4, and two of 46 (4%) with CEA. Of the lung adenocarcinomas, 18 of 20 (90%) were positive with MOC 31, 20 of 20 (100%) with Ber-EP4, and 17 of 20 (85%) with CEA; and two of 20 (10%) were focally reactive with calretinin, one of 20 (5%) with thrombomodulin, none of 20 (0%) with CKs 5/6, and five of 20 (25%) with high-weight CKs. Multiple logistic modeling indicated two batteries of three antibodies permitting more than 98% overall accuracy: Ber-EP4 plus CKs 5/6 plus calretinin, and Ber-EP4 plus CKs 5/6 plus CEA.

Topics: Adenocarcinoma; Antibodies; Antigens, Neoplasm; Antigens, Surface; Biomarkers, Tumor; Calbindin 2; Carcinoembryonic Antigen; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Mesothelioma; Regression Analysis; S100 Calcium Binding Protein G; Thrombomodulin

The CYFRA 21-1 assay, which detects cytokeratin 19 (CK19) fragment, is widely used as a tumor marker for lung cancer, particularly non-small cell lung cancer. However, the reason that some lung cancer cell lines release CYFRA 21-1 in culture supernatants and others do not remains unclear. We hypothesized that the release of CYFRA 21-1 might be related to the expression of CK19 and caspase 3. In order to prove this, the quantities of mRNA for CK19 were evaluated by the competitive reverse transcriptase-polymerase chain reaction (RT-PCR). CK19 protein synthesis was also evaluated by Western blotting and immunohistochemistry, and the levels of CYFRA 21-1 in the culture supernatant were measured by an immunoradiometric assay. The expression of mRNA for caspase 3 was evaluated by the RT-PCR, and caspase 3 protein synthesis was also evaluated by immunohistochemistry. In 13 lung cancer cell lines, the amounts of mRNA for CK19 correlated with the levels of CYFRA 21-1 in culture supernatants, results of Western blotting for CK19, and positivities of immunohistochemistry for CK19. In 5 cell lines that produced a significant amount of CYFRA 21-1, the level of CYFRA 21-1 correlated with the positivity of RT-PCR for caspase 3 and immunohistochmistry for caspase 3. This suggests that caspase 3 played a role in the formation of CYFRA 21-1. In addition, the specific inhibitor of caspase 3 significantly inhibited the release of CYFRA 21-1 in culture supernatants. In conclusion, we demonstrate that caspase 3, which cleaves several intermediate filaments and carries out cell apoptosis, played an important role in producing CYFRA 21-1 in human lung cancer cell lines.

Topics: Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Apoptosis; Biomarkers, Tumor; Blotting, Western; Caspase 3; Caspase 6; Caspases; Electrophoresis, Polyacrylamide Gel; Humans; Immunohistochemistry; Keratin-19; Keratins; Lung Neoplasms; Mice; Models, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Time Factors; Tumor Cells, Cultured

Metastatic carcinoma of unknown primary is a common problem, accounting for up to 10-15% of all solid tumours at presentation. Proper identification of the site of origin has prognostic and therapeutic significance. Prior immunohistochemical methods to identify the site of origin have been useful in a limited number of cases. Differential cytokeratin staining may be useful in this setting, particularly in identifying metastases from lung cancer. We have identified 144 cases of metastatic carcinoma of unknown primary to bone, lung or liver at Brigham and Women's Hospital between 1 January 1997 and 1 July 1998. Cytokeratin (CK) 7 and CK20 were used in 75 of these cases to narrow down the possible sites of the primary tumours. All of these cases were ambiguous as to the site of the primary tumour. Forty-five cases were CK7+/CK20-, 15 cases were CK7-/CK20-, 9 cases were CK7-/CK20+ and 6 cases were CK7+/CK20+. Three of the cases were selected for detailed presentation and discussion as well as a discussion of the pertinent literature. Overall, the CK7+/CK20- phenotype favours a lung primary, the CK7+/CK20+ phenotype strongly favours transitional cells (urothelial) carcinoma, the CK7-/CK20+ phenotype favours colorectal carcinoma, while the CK7-/CK20- profile is not helpful.

Topics: Biomarkers; Bone Neoplasms; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Liver Neoplasms; Lung Neoplasms; Neoplasms, Unknown Primary

Numerous epidemiologic studies suggest a relationship between lung cancer and peptic ulcer disease. Furthermore, various lung cancers synthesize and release a number of peptides such as gastrin and gastrin-releasing peptide that could cause acid hypersecretion; however, Zollinger-Ellison syndrome (ZES), because of a lung tumor, has never been described. We report such a patient for the first time. A 60-year-old man with a non-small cell lung carcinoma (large cell type) presented with diarrhea, heartburn, abdominal pain, and duodenal ulcers. Evaluation showed ZES was present (fasting hypergastrinemia, hyperchlorhydria) and control of all symptoms by omeprazole. No abdominal or cardiac tumor, the other known locations of gastrinomas causing ZES, was found on detailed tumor imaging studies. Resection of the lung tumor resulted in a decrease in gastrin levels to normal values. Plasma radioimmunoassays showed elevated gastrin, chromogranin A and normal levels of gastrin-releasing peptide, and 9 other hormones. The tumor showed similar immunocytochemical results. The characteristics of this case are compared with 100 cases of sporadic abdominal gastrinomas, and the evidence reviewed suggests why ZES should be considered in patients with lung cancer with peptic symptoms.

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Chromogranin A; Chromogranins; Gastric Acid; Gastrins; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Synaptophysin; Zollinger-Ellison Syndrome

We assessed the association of vascular endothelial growth factor (VEGF) and nm23 expression with occult micrometastasis in lung cancer. As destination sites for micrometastasis, we scrutinized lymph node (LN) and bone marrow (BM) specimens. For LN, 122 stage I patients who had received curative operations were studied. As regards BM, 203 patients in stage I - IV who underwent operations were registered. Immunohistochemical anti-cytokeratin staining was used to detect microdissemination of cancer cells. The VEGF and the nm23 expression at the primary sites were immunohistochemically studied in 285 cases in total. The percentages of the patients with microdissemination were 28.7% for LN and 42.4% for BM. The outcome for the patients with LN or BM microdissemination was significantly worse than that for patients without it. The increased VEGF and the decreased nm23 expression within primary tumors were significantly associated with LN and BM microdissemination. The results indicate possible value of using these biological markers to predict the risk of systemic micrometastasis in non-small cell lung cancer.

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; Bone Marrow; Carcinoma, Adenosquamous; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Endothelial Growth Factors; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Lymphokines; Male; Middle Aged; Monomeric GTP-Binding Proteins; Neoplasm Metastasis; Neoplasm Staging; NM23 Nucleoside Diphosphate Kinases; Nucleoside-Diphosphate Kinase; Predictive Value of Tests; Sensitivity and Specificity; Transcription Factors; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Lung adenocarcinomas are heterogeneous clinically and histologically. Expression of the mucin genes was analyzed as a molecular marker of glandular cytodifferentiation in primary lung adenocarcinomas. Expression was correlated with histopathologic subtypes of World Health Organization classification with the aim of investigating the histogenesis of primary lung adenocarcinomas. Thirty-four primary lung adenocarcinomas were examined by in situ hybridization for mucin gene expression (MUC1-4, MUC5AC, MUC5B, MUC6-7) and by immunohistochemistry for MUC5AC and MUC5B apomucin expression. Mucinous bronchioloalveolar carcinoma (BAC) had a homogeneous pattern of mucin gene expression different from those of other types of lung adenocarcinoma, involving secreted mucins (MUC5AC, MUC5B, and MUC6) and membrane-bound mucins (MUC1, MUC3, and MUC4). Non-BAC adenocarcinoma and mucinous BAC aberrantly expressed mucin genes MUC3, and MUC3 and MUC6, respectively, which are undetectable in normal fetal and adult lung. Our results show the particular phenotype of mucin gene expression in mucinous type of BACs and the heterogeneous expression of respiratory and nonrespiratory mucins in the other types. This finding supports the theory of a common progenitor cell with the potential of multicellular differentiation. From a practical point of view, the aberrant expression of MUC3 and MUC6 could serve as a diagnostic marker in the management of the mucinous type of bronchioloalveolar carcinomas. HUM PATHOL 32:274-281.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Adult; Aged; Female; Gene Expression; Histocytochemistry; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Lung Neoplasms; Male; Middle Aged; Mucins; RNA, Messenger

The aim of our study was to assess the clinical value of CYFRA 21-1 tumor marker and carcinoembryonic antigen (CEA) as diagnostic tools that are complementary to cytology in the diagnosis of malignant mesotheliomas.. We measured CEA and CYFRA 21-1 in the pleural effusions (PEs) and serum of 106 patients (benign lung disease, 34 patients; bronchogenic and metastatic carcinoma, 40 patients; mesothelioma, 32 patients).. CEA and CYFRA 21--1 levels were determined by means of two commercial enzyme immunoassays.. The cutoff levels of CYFRA 21--1 and CEA in malignant PEs, selected on the basis of the best diagnostic efficacy, were 41.9 ng/mL and 5.0 ng/mL, respectively. In all neoplastic PEs, CYFRA 21--1 and CEA sensitivity was 78% and 30.6%, respectively, with a specificity of 80% and 91%, respectively. The sensitivity of CYFRA 21--1 and CEA in patients with mesothelioma was 87.5% and 3.1%, respectively. The results of the CYFRA 21--1 assay were positive in 17 of 19 cases of mesothelioma (89.5%) with a negative or uncertain cytology. The association of the tumor marker assay and the cytology allowed a correct diagnosis in 30 of 32 cases of mesothelioma (93.7%).. This study suggests that CYFRA 21--1 would provide a useful parameter for the differential diagnosis between benign and malignant PE from mesothelioma when the result of cytology is negative or uncertain and the clinical context does not allow a more aggressive approach. Moreover, the association of CYFRA 21--1 with CEA could provide details for a differential diagnosis between mesotheliomas and carcinomas. In fact, an elevated CYFRA 21--1 level with a low CEA level is highly suggestive of mesothelioma, whereas high levels of CEA alone or high levels of both the markers suggest a diagnosis of malignant PE, excluding mesothelioma.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma; Diagnosis, Differential; Humans; Keratin-19; Keratins; Lung Diseases; Lung Neoplasms; Mesothelioma; Pleural Effusion, Malignant; Pleural Neoplasms; Sensitivity and Specificity

The authors report a case of gangliocytic paraganglioma of the lung, which has not yet been described in a pulmonary neoplasm. A 75-year-old man underwent right middle and lower lobe lobectomy. A slightly yellowish mass was located at the bifurcation between the lower and middle lobe bronchus, protruding into the truncus intermedius. The neoplastic cells were composed of three cellular elements: uniform endocrine cells in a Zellballen arrangement, large ganglion-like cells within the nests of endocrine cells, and spindle-shaped cells arranged in streams to surround the nests. Each component exhibited the characteristic immunohistochemical properties, which were similar to those of the corresponding neuroendocrine neoplasms: Endocrine cells were positive for CAM 5.2, chromogranin A, and synaptophysin, like carcinoid tumor; ganglion-like cells were positive only for neurofilament, like ganglioneuroma; and spindle-shaped cells were positive for neurofilament and S-100 protein, like paraganglioma. These results agreed with those in gangliocytic paraganglioma of the duodenum. Pulmonary gangliocytic paraganglioma is similar to that in the duodenum, and is a hamartomatous proliferation of epithelial endocrine and neuronal cells of the bronchus.

Topics: Aged; Biomarkers; Biomarkers, Tumor; Chromogranin A; Chromogranins; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Neoplasm Proteins; Neurofilament Proteins; Paraganglioma; Radiography, Thoracic; S100 Proteins; Synaptophysin; Tomography, X-Ray Computed

histological features of metastasis are not always specific of the origin. The usefulness of cytokeratin 7 and 20 (CK7 and CK20) in cerebral metastases from an adenocarcinoma was studied in a series of 78 patients.. metastases from lung adenocarcinoma showed a CK7+/CK20- pattern in 79% of cases, CK7+/CK20+ in 19% and CK7-/CK20- in 2%. When observed, positivity for cytokeratin 20 was generally focal or weak. Breast adenocarcinoma metastases were CK7+/CK20- in 71% of cases and CK7-/CK20- in 29%. Less differentiated tumours were usually negative for both cytokeratins. Metastases by colonic adenocarcinoma were CK7-/CK20+ in 100% of cases. Cytokeratins 7 and 20 are useful to distinguish lung from colonic metastatic carcinoma. In case of double positivity, a more intense CK7 expression is rather suggestive of a lung origin.. these results might modify paraclinic investigations in search of the primitive tumor.

Topics: Adenocarcinoma; Brain Neoplasms; Breast Neoplasms; Colonic Neoplasms; Diagnosis, Differential; Humans; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms

The distinction between pleural epithelial mesothelioma and peripheral lung adenocarcinoma involving the pleura is still an important diagnostic problem for surgical pathologists. The aim of our study was to identify the most specific and sensitive markers for the positive identification of mesothelioma to select a limited, appropriate panel of antibodies to differentiate between mesothelioma and adenocarcinoma. Forty-two cases of epithelial mesotheliomas and 23 cases of pulmonary adenocarcinomas were stained with the following antibodies: anticalretinin, antithrombomodulin, anti-CD44H, and monoclonal antibody HBME-1. We also studied the value of other markers in current use: cytokeratins AE1/AE3 and CAM5.2, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), Ber-EP4, B72.3, and CD15. Of the mesotheliomas, 42 stained for calretinin, 39 (92.8%) for thrombomodulin, 42 stained for CD44H, and 41 (97.6%) stained for HBME-1. Among negative markers, 4 (9.5%) mesothelioma cases stained for CEA, 5 (11.9%) stained for Ber-EP4, 6 (14.2%) stained for B72.3, and 2 (4.7%) stained for CD15. Of the lung adenocarcinomas, 2 (8.7%) cases showed reactivity for calretinin, 5 (21.7%) for thrombomodulin, 13 (56.5%) for CD44H, all for HBME-1, 22 (95.6%) for CEA, 22 (95.6%) for Ber-EP4, 8 (34.7%) for B72.3, and all for CD15. In conclusion, calretinin and thrombomodulin were the most specific positive mesothelial markers, whereas CD44H and HBME-1 showed high sensitivity but very low specificity. Among negative markers, we advocate the use of CEA and CD15 which were the most specific in differentiating mesotheliomas from adenocarcinomas.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Biomarkers, Tumor; Calbindin 2; Carcinoembryonic Antigen; Diagnosis, Differential; Humans; Hyaluronan Receptors; Immunohistochemistry; Keratins; Lewis X Antigen; Lung Neoplasms; Mesothelioma; Mucin-1; Pleural Neoplasms; S100 Calcium Binding Protein G; Thrombomodulin

Although numerous studies have shown that nm23-H1 gene product expression is inversely related to metastatic potential in some cancers, the expression in lymph nodes has not been studied in detail. An analysis of nm23-H1 gene product expression in mediastinal lymph nodes from lung cancer patients is reported.. One hundred and thirty-four, randomly selected lymph nodes (63 with positive pathological lymph node status) from 39 surgically treated lung cancer patients were examined. Expression of nm23-H1 gene product was determined using specific monoclonal antibodies. Metastatic cancer cells were highlighted using anti-cytokeratin antibody.. Expression of nm23-H1 gene product in patients with less and more than 50% nodes-positive was 12/23 (52.2%) and 15/16 (93.8%) cases, respectively. Immunohistochemical studies with cytokeratin revealed micrometastasis in 6/39 (15.4%) patients and 9/71 (12.7%) nodes previously reported as cancer negative. Expression of nm23-H1 gene product in micrometastasis and metastasis-positive nodes was 5/9 (55.6%) and 55/63 (87.3%), respectively. We also found nm23-H1 gene product expression in germinal center cells. However, we found no relationship between expression of nm23-H1 gene product in germinal center cells and extent of metastasis.. Our study demonstrates a positive relationship between expression of nm23-H1 gene product and extent of metastasis in mediastinal lymph nodes from lung cancer patients. Our data for normal germinal center cells suggests that nm23-H1 gene product expression does not play a specific biological role in suppressing tumor metastasis in lung cancer.

Topics: Adenocarcinoma; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Mediastinum; Monomeric GTP-Binding Proteins; NM23 Nucleoside Diphosphate Kinases; Nucleoside-Diphosphate Kinase; Transcription Factors

We determined the clinicopathological features of primary lung carcinomas with rhabdoid cells by defining the immunophenotype of rhabdoid cells and analysing survival.. Rhabdoid cells are distinctive in having an eccentric nucleus and a large intracytoplasmic inclusion on routinely stained sections. Based on the number of rhabdoid cells, 45 cases of large cell carcinoma were divided into the following three types: lung tumour with a rhabdoid phenotype (LTRP) (n=4), lung carcinoma with a small number of rhabdoid cells (LCSR) (n=10), large cell carcinoma containing no rhabdoid cells (LCNR) (n=31). LTRP is composed of at least 10% rhabdoid cells. In LCSR the percentage of rhabdoid cells is less than 10%. LTRP and LCSR are associated with locally advanced disease. Immunohistochemical stains were positive for epithelial markers in all LTRP and eight LCSR, for neuroendocrine markers in one LTRP and three LCSR. The outcome is worse for patients with LTRP than LCSR or LCNR. LCSR shows a trend close to LCNR. Stage-matched survival analysis, however, revealed no statistically significant difference among the histological subtypes.. Rhabdoid cells are heterogeneous except for epithelial markers and vimentin positivity. Less than 5% of rhabdoid cells has a negligible effect on prognosis.

Topics: Adult; Aged; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Microscopy, Electron; Middle Aged; Mucin-1; Neoplasm Staging; Phosphopyruvate Hydratase; Rhabdoid Tumor; Survival Analysis; Vimentin

In cytological preparations, reactive mesothelial cells (RMC) in serous effusions are sometimes difficult to distinguish from adenocarcinoma cells (AC). RMC and AC can be distinguished by lectin-binding patterns, but the pattern of binding of lectins to normal mesothelium is not well defined. We investigated the expression of cytoskeletal filaments, cytokeratin (CK) and vimentin (VM), and the cell surface binding pattern of 10 lectins (HPA, SBA, ABA, DSA, PNA, RCA-I, UEA-I, LTA, WGA and ConA) in the serosa of 48 adenocarcinoma specimens. We also investigated the usefulness of six lectins (HPA, SBA, RCA-I, UEA-I, LTA and WGA) in identification of RMC and AC in 16 serous effusions. DSA reactivity was significantly higher (P < 0.05) in static mesothelial cells (SMC) than in RMC. Reactivity for LTA and ConA was significantly lower (P < 0.05) in SMC than in RMC. Anti-CK and anti-VM immunoreactivity was always positive in RMC and almost negative in SMC. In serous effusions, HPA, SBA and UEA-I binding was evident in 100, 88 and 81% of AC, respectively. Little to no binding of HPA, SBA or UEA-I was detected in RMC. Our results suggest that the morphological differences between SMC and RMC are likely to be due to differences in cytoskeletal composition, with accompanying changes in cell-surface lectin-binding patterns. HPA, SBA and UEA-I are likely to be useful markers for identification of RMC and AC in cytology.

Topics: Adenocarcinoma; Ascites; Binding Sites; Biomarkers; Cytoskeleton; Epithelium; Gastrointestinal Neoplasms; Humans; Immunohistochemistry; Keratins; Lectins; Lung Neoplasms; Pleural Effusion; Serous Membrane; Vimentin

To report about a lung tumor that was a combination of typical carcinoid and adenocarcinoma.. The patient, a 71-year-old male, presented with a 2.5-cm pulmonary nodule that, microscopically, was a combination of an adenocarcinoma (tubular with clear cell features and bronchioloalveolar) and a typical carcinoid. Immunohistochemically, both components were positive for cytokeratin, but only the carcinoid component was positive for chromogranin and synaptophysin. In the range of neuroendocrine tumors of the lung, a combination with other histological types of carcinoma (squamous, adeno, large cell and pleomorphic) can be found with both small cell carcinoma and large cell neuroendocrine carcinoma, but is very rare with typical and atypical carcinoids.

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Adenocarcinoma, Clear Cell; Aged; Biomarkers, Tumor; Carcinoid Tumor; Carcinoma, Non-Small-Cell Lung; Chromogranins; Humans; Keratins; Lung Neoplasms; Male; Neoplasm Proteins; Neoplasms, Multiple Primary; Synaptophysin

We have investigated the serum level of granulocyte-colony stimulating factor (G-CSF) and macrophagecolony stimulating factor (M-CSF) in non-small-cell lung cancer (NSCLC), in relation to the control group and commonly accepted tumor markers, such as carcinoembryonic antigen (CEA) and cytokeratin fragment 19 (CYFRA 21-1). Additionally, we have defined the diagnostic sensitivity, specificity, positive predictive value, negative predictive value and receiver-operating characteristics (ROC) curve of G-CSF and M-CSF. Serum levels of cytokines were measured in 61 patients with NSCLC and in 20 healthy subjects. G-CSF and M-CSF were determined using ELISA. CYFRA 21-1 was measured by radioimmunoassay and CEA by microparticle enzyme immunoassay. There were significant increases in the level of circulating G-CSF in the lung cancer patients compared to the control group. Moreover, the diagnostic sensitivity of G-CSF was higher (56%) than the sensitivity of CYFRA 21-1 (51%), but lower than the CEA sensitivity (62%). The diagnostic specificity of G-CSF was higher (70%) than the M-CSF specificity (40%) and the G-CSF predictive values were higher in relation to the predictive values of M-CSF. These results suggest a potential role of G-CSF as a tumor marker for NSCLC.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Enzyme-Linked Immunosorbent Assay; False Negative Reactions; Female; Granulocyte Colony-Stimulating Factor; Humans; Keratin-19; Keratins; Lung Neoplasms; Macrophage Colony-Stimulating Factor; Male; Middle Aged; Predictive Value of Tests; Radioimmunoassay; ROC Curve; Sensitivity and Specificity

Tumor cells generally present various types of nuclear alterations, which can be associated with genetic instability. The origin and mechanism of formation of the nuclear alterations are largely unknown, with the micronucleus being the most well studied alteration. The purpose of this study was to characterize the cytoskeleton filaments and to analyze the possible association between nuclear alterations and the cytoskeleton in the human lung carcinoma cells HK2 and A549. The cytoskeleton analysis was performed by using antibodies against lamin B, vimentin, cytokeratin-8, and alpha-tubulin and the secondary antibody labeled with FITC. The analysis of the actin filament was made with phalloidin-TRITC. The analyses of cytoskeleton were performed from optical sections obtained by confocal laser scanning microscopy. Filaments of the cytoskeleton of tumor cells present some differences in their distribution pattern and their expression when compared with the filaments of normal cells. The HK2 cells presented actin fibers arranged either concentrically or in clusters and tubulin filaments arranged radially, while in the A549 cells the distribution pattern was similar to that of normal cells. The lamin B filaments were the most important to identify nuclear alterations. These alterations in cytoskeleton distribution could not be associated with nuclear alterations.

Topics: Actin Cytoskeleton; Actins; Animals; Cell Nucleus; Cytoskeleton; Humans; Keratins; Lung Neoplasms; Microscopy, Confocal; Rats; Tubulin; Tumor Cells, Cultured; Vimentin

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Clinical Trials as Topic; Humans; Keratin-19; Keratins; Lung Neoplasms; Prognosis; Reproducibility of Results; Treatment Outcome

Spindle cell carcinoma of the breast, a variant of metaplastic carcinoma, includes a wide spectrum of lesions with histomorphologic and nuclear features ranging from overtly malignant to mildly atypical. Spindle cell carcinomas with mildly atypical features may resemble fasciitis, fibromatosis, or myofibroblastic tumors and therefore are often misinterpreted as such. A recent study has suggested that spindle cell carcinomas with a dominant fibromatosis-like phenotype, unlike spindle cell carcinomas in general, have no propensity for distant metastasis and should be termed "tumors" rather than "carcinomas." To investigate the question of fibromatosis-like spindle cell breast carcinoma (FLSpCCs) metastatic potential, we studied cases of FLSpCC seen at the University of Texas M.D. Anderson Cancer Center between 1987 and 2000. Clinical, pathologic, and immunophenotypic features were reviewed, with emphasis on biologic behavior and predictors of clinical outcome. Our series included 24 women who ranged in age from 55 to 85 years (mean 66 years). Tumor size ranged from 1.0 to 5 cm (mean 2.8 cm). Most tumors were grossly well defined but had microscopic infiltrative borders. Tumors showed a dominant fibromatosis-like or myofibroblastic-like growth pattern with prominent collagenization. Inflammatory infiltrate was noted in the majority of tumors. Cytokeratin-positive cells were seen in all cases and usually appeared as cords or sheets of polygonal cells; isolated cytokeratin-positive cells were rare. In most tumors immunoreactivity for smooth muscle actin (SMA) was confined to the cytokeratin-negative cells. In five cases intense co-expression of cytokeratin and SMA was noted. None of the tumors showed immunoreactivity for smooth muscle heavy chain myosin, estrogen receptors, progesterone receptors, or HER-2/neu. Ki-67 expression was noted in fewer than 5% of tumor cells. Treatment consisted of local excision (seven cases) or modified radical mastectomy (13 cases). Treatment was unknown in four cases. In patients who underwent axillary nodal dissection, no lymph node metastases were found. Two of the six patients who underwent local excision developed local recurrence. Two patients who underwent modified radical mastectomy developed lung metastases within 2 years after the initial diagnosis. The metastatic tumors were histologically similar to the primary tumors. Our findings indicate that FLSpCCs have the potential for local recurrence and distant metastasis an

Topics: Actins; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Squamous Cell; Female; Fibroma; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Middle Aged; Neoplasm Recurrence, Local

Our purpose was to determine whether peripheral blood biomarkers MUC1 and CK19 could be used to complement imaging studies in differentiating benign from malignant indeterminate pulmonary nodules or masses detected on computed tomography CT. One hundred and eighteen patients had a thoracic CT and blood drawn for tumor marker reverse transcriptase-polymerase chain reaction analysis. Thirty-five of the 118 patients had an indeterminate pulmonary nodular opacity on CT, and the findings then were correlated with the reverse transcriptase-polymerase chain reaction results. The sensitivity and specificity for the markers in determining malignancy was calculated. Thirteen of the 35 opacities on CT proved to be benign, and 22 proved to be lung cancer. Among the patients with indeterminate pulmonary abnormalities, polymorphic epithelial mucin protein 1 had a sensitivity and specificity for lung cancer of 100% and 46%, respectively. Cytokeratin 19 had a sensitivity and specificity for lung cancer of 95% and 8%, respectively. These preliminary data showed that serum biomarkers polymorphic epithelial mucin protein 1 and cytokeratin 19 were not specific for lung cancer, although patients with an indeterminate pulmonary abnormality and negative markers were unlikely to have lung cancer. Integration of imaging studies with the appropriate biomarkers may prove useful in evaluating indeterminate pulmonary nodules or masses.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Diagnosis, Differential; Female; Humans; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Mucin-1; Peptide Fragments; Pilot Projects; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tomography, X-Ray Computed

We studied 14 mucinous and 26 nonmucinous bronchioloalveolar adenocarcinomas (BACs) with thyroid transcription factor (TTF), cytokeratin (CK) 7, CK20, and villin to characterize their staining patterns with these antibodies and identify staining differences between the neoplasms. We also stained 11 mucinous colon adenocarcinomas with the same antibodies to compare their reaction patterns with mucinous BACs. All pulmonary neoplasms were confirmed pulmonary primary BACs. Three (21%) of 14 mucinous neoplasms had weak TTF reactivity in fewer than 25% of neoplastic cell nuclei, and the other 11 (79%) were nonreactive. In contrast, 24 (92%) of 26 nonmucinonus BACs were strongly TTF reactive. Eleven mucinous BACs (79%) had CK20 reactivity in more than 25% of neoplastic cells, whereas only 1 nonmucinous BAC (4%) had reactivity in fewer than 50% of the cells. One mucinous BAC (7%) had villin reactivity in approximately 10% of the neoplastic cells. All mucinous colon adenocarcinomas were diffusely reactive with CK20 and villin. Mucinous and nonmucinous BACs have disparate staining patterns with TTF and CK20. Mucinous BACs are usually TTF nonreactive and CK20 reactive, but nonreactive with villin, which distinguishes them from mucinous colon adenocarcinomas.

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Adenocarcinoma, Mucinous; Biomarkers, Tumor; Carrier Proteins; Cell Nucleus; Colonic Neoplasms; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Microfilament Proteins; Nuclear Proteins; Staining and Labeling; Thyroid Nuclear Factor 1; Transcription Factors

Basaloid carcinoma (BC) and large-cell neuroendocrine carcinoma (LCNEC) are 2 recently recognized variants of large-cell lung carcinomas that may overlap in their morphology, and are discriminated by expression of neuroendocrine markers in LCNEC. Because thyroid transcription factor 1 (TTF-1) is expressed in lung adenocarcinomas but not in squamous cell carcinomas (SCC), and 34betaE12 recognizes a set of high-molecular-weight cytokeratins characteristic of basal stem cells, we hypothesized that these 2 markers could help in distinguishing BC from LCNEC. Immunostaining for TTF-1 was detected in 40.9% of pure LCNEC but in no BC or basaloid variant of SCC. In contrast, immunoreactivity for 34betaE12 was shown in all BC and basaloid variant of SCC but in only 1 LCNEC. Bouin fixation was less efficient than formalin in the immunodetection of both markers for its well-known deleterious effect on antigen preservation. Specificity of TTF-1 for LCNEC (100%) and that of 34betaE12 for BC (98.3%) exceeded that of NE markers for distinction of these 2 entities. These data show that TTF-1 and 34betaE12, in association with specific neuroendocrine markers, represent a useful panel of antibodies in differentiating carcinomas presenting with a solid pattern, palisading, or pseudorosettes, the expression of TTF-1 excluding the diagnosis of BC, and staining with 34betaE12 excluding pure LCNEC.

Topics: Carcinoma, Large Cell; Carcinoma, Neuroendocrine; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Nuclear Proteins; Retrospective Studies; Sensitivity and Specificity; Thyroid Nuclear Factor 1; Transcription Factors

Sclerosing hemangioma of the lung is well characterized histologically, but the line of differentiation expressed by the tumor cells has been unclear. Despite the implication by its name of a vascular neoplasm, sclerosing hemangioma is considered by most authorities to be an epithelial tumor, possibly related to the pulmonary epithelium.. To determine the line of differentiation of the tumor cells with immunohistochemistry and to review the related literature.. Nine cases of histologically typical pulmonary sclerosing hemangioma were studied with pan-epithelial (epithelial membrane antigen [EMA] and CAM 5.2), endothelial (CD31), neuroendocrine (chromogranin A), and pulmonary epithelial markers (thyroid transcription factor-1 and PE10). Staining intensity was separately evaluated in the pale cells of the solid areas and the cells lining the papillary structures.. Both cell types were positive for thyroid transcription factor-1 and EMA in all cases (100%). Thyroid transcription factor-1 showed diffuse strong staining, and EMA staining varied from focal weak to diffuse strong. The pale cells showed focal staining for keratin (CAM 5.2) in 2 (28%) of 7 cases, and for PE10 in 5 (62%) of 8 cases. The papillary lining cells were at least focally positive with CAM 5.2 and PE10 in all cases (100%). Reactions for chromogranin and CD31 were negative in both cell types in every case. The number of PE10- or CAM 5.2-positive papillary lining cells was less than the number of EMA-positive papillary lining cells.. The uniform positivity for EMA is consistent with the notion that the tumor cells of sclerosing hemangioma are epithelial, and the strong thyroid transcription factor-1 positivity suggests differentiation toward pulmonary epithelium. The papillary lining cells expressing EMA as well as PE10 or CAM 5.2 likely represent entrapped metaplastic alveolar epithelium, whereas the papillary lining cells expressing only EMA more likely constitute true neoplastic cells similar to those in the solid areas.

Topics: Adult; Aged; Biomarkers; Biomarkers, Tumor; Chromogranin A; Chromogranins; Female; Hemangioma; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Mucin-1; Nuclear Proteins; Platelet Endothelial Cell Adhesion Molecule-1; Protein Precursors; Proteolipids; Thyroid Nuclear Factor 1; Transcription Factors

Evaluation of prognosis-associated parameters in patients with small cell lung carcinoma.. Biopsies of 46 patients suffering from a non-treated small cell lung carcinoma were stained with Feulgen and immunohistochemically with Ki-67 antibody. The integrated optical density (IOD) and proliferation rate was measured by syntactic structure analysis and correlated with survival.. About 85% of patients had a smoking history (46 pack years on average). The median survival time was 13.5 months, the proliferation rate (Ki-67 positive tumor cell nuclei) 68.2% and S-phase percentage 9.2%. Ot average, 25 proliferating tumor cell nuclei formed clusters (mean diameter 95 microns). The prognosis was associated with the proliferation rate (p < 0.04), tumor stage (stage I versus lib, p < 0.05), at threshold limits with S-phase rate (p < 0.07) and serum levels of LDH and NSE (p < 0.06 and p < 0.07 respectively).. Immune histochemical determination of the Ki-67 protein is a useful method to estimate the prognosis of patients with small cell lung carcinoma.

Topics: Antigens, Neoplasm; Biopsy; Carcinoembryonic Antigen; Carcinoma, Small Cell; Female; Follow-Up Studies; Histocytochemistry; Humans; Immunohistochemistry; Keratin-19; Keratins; Ki-67 Antigen; L-Lactate Dehydrogenase; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Prognosis; Retrospective Studies; Smoking; Survival Rate; Time Factors

The distinction of a primary lung carcinoma from a metastatic lesion is important, because the treatment and prognosis differ for patients with these malignancies. Such a distinction can be difficult because of overlapping cytologic features. It has been shown that antibodies to thyroid transcription factor 1 (TTF-1) and PE-10 are fairly specific markers for primary lung tumors in histologic specimens. TTF-1 regulates the expression of surfactant protein production, and PE-10 is a monoclonal antibody against components of human surfactant proteins. The combination of cytokeratin 7 (CK7) and cytokeratin 20 (CK20) immunoprofiling has been helpful in the identification of the primary site of origin of lung tumors.. In the current study, the authors evaluated the utility of TTF-1 and PE-10 immunostaining and also compared the staining with expression of CK7 and CK20 in the discrimination between primary lung tumors and metastatic lesions in 55 specimens from fine-needle aspiration (FNA) biopsies of the lung. Formalin fixed, paraffin embedded cell blocks from 35 primary lung tumors (16 adenocarcinomas, 8 squamous cell carcinomas, 6 large cell undifferentiated carcinomas, and 5 small cell carcinomas) and 20 metastatic carcinomas (6 breast lesions, 6 colon lesions, 3 urinary bladder lesions, 2 kidney lesions, 1 biliary tract lesion, 1 endometrial lesion, and 1 thyroid lesion) were immunostained with monoclonal antibodies to TTF-1, PE-10, CK7, and CK 20. Positive immunostaining for CK7, CK20, and PE-10 was based on cytoplasmic staining, whereas TTF-1 positive staining was based on nuclear staining of the neoplastic cells.. Positive immunostaining with TTF-1 and PE-10 was noted in six primary lung tumors (17%). One metastatic lesion (5%) and two metastatic lesions (10%) were positive for TTF-1 and PE-10, respectively. The CK7 positive/CK20 negative immunophenotype was noted in 30 primary lung tumors (86%) and in 11 metastatic lesions (55%). The CK7 negative/CK20 negative immunophenotype was seen in four metastatic lesions and in the remaining five primary lung tumors. The CK7 negative/CK20 positive and CK7 positive/CK20 positive immunophenotypes were seen in two and three metastatic lesions, respectively, but in none of the primary lung tumors. When a CK7 positive/CK20 negative adenocarcinoma also demonstrated either TTF-1 positive or PE-10 positive staining, it was likely that the adenocarcinoma was of pulmonary origin (P < 0.035; Fisher exact test). The specificity of such a combination for discriminating between primary and metastatic adenocarcinomas was 94%.. The results suggest that TTF-1, PE-10, or CK7/CK20 alone did not distinguish reliably between primary pulmonary tumors carcinomas and metastatic neoplasms of the lung in FNA biopsy specimens because of low sensitivity and specificity. The use of a panel of antibodies that includes CK7/CK20, TTF-1, and PE-10 may be helpful in discriminating between primary and metastatic adenocarcinomas of the lung. An adenocarcinoma is likely a primary lung tumor when it is of the CK7 positive/CK20 negative phenotype and demonstrates either TTF-1 positive or PE-10 positive staining.

Topics: Adenocarcinoma; Biomarkers, Tumor; Biopsy, Needle; Carcinoma, Large Cell; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Neoplasm Metastasis; Nuclear Proteins; Predictive Value of Tests; Pulmonary Surfactants; Sensitivity and Specificity; Thyroid Gland; Thyroid Nuclear Factor 1; Transcription Factors

Keratins form the largest subfamily of intermediate filament proteins and show strict lineage- and differentiation-associated expression in epithelial cells. Little is known about the mechanisms that control keratin protein synthesis in these cells. We have examined the effect of the differentiation-modulating agent, 5'-bromo-2'-deoxyuridine (BrdU), on keratin 19 (K19) expression in two human lung carcinoma cell lines, DLKP and A549. Treatment of both cell lines with 10 microM BrdU for 7 d induced the expression of K19 protein in keratin-negative DLKP cells, and significantly increased K19 protein expression in A549 cells. K19 messenger ribonucleic acids (mRNAs) were detected by Northern blot and reverse transcriptase-polymerase chain reaction analyses in both cell lines, but no increase in K19 mRNA levels was detected in either cell line, even with treatment with BrdU for up to 21 d. This suggests that K19 protein synthesis is normally blocked at a posttranscriptional level in DLKP cells, and BrdU can somehow reverse this block, resulting in the induction of K19 protein synthesis. Treatment of HL60, a leukemic cell line, with BrdU, resulted in noninduction of K19 protein synthesis, and no K19 mRNA transcripts were detected before and after BrdU treatment, possibly suggesting that BrdU is acting in an epithelial-specific manner to reverse a block in K19 protein synthesis in DLKP keratin-negative lung cancer cells. Therefore, DLKP (and A549) may be useful cellular models to investigate if this represents a regulatory step in early lung development or a mechanism whereby tumor cells possess the ability to down-regulate the expression of a more-differentiated phenotype.

Topics: Blotting, Northern; Bromodeoxyuridine; Gene Expression; HL-60 Cells; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured

Expression of cytokeratin (CK) 7 and 20 is commonly used to help distinguish adenocarcinomas from different sites. Thyroid transcription factor 1 (TTF-1) is a 38-kd protein, located primarily in the nucleus of type 2 pneumocytes and clara cells. TTF-1 has been shown to be present in a variety of lung and thyroid tumors and in pulmonary small-cell carcinomas. Carcinoid tumors from the lung and the gastrointestinal (GI) tract are histologically similar and thus are difficult to differentiate from each other based on histologic criteria. Pancreatic endocrine tumors (PET) have a similar histologic appearance to these other tumors. The purpose of this study was to determine the efficacy of differentiating these 3 groups of tumors by their expression of CK7, CK20, and TTF-1. Routinely processed paraffin-embedded tissue sections from 62 carcinoid tumors (lung, 16; gastrointestinal [GI] tract, 46) and 12 PETs were immunohistochemically stained for CK7, CK20, and TTF-1. The degree of expression in each tumor was graded as 1+ (1% to 10% of cells positive), 2+ (11% to 25%), 3+ (26% to 50%), and 4+ (>50%). The data were compared between tumor types and between carcinoid tumors from the various locations in the GI tract (stomach, 8; small intestine, 19; large intestine, 17; appendix, 2). CK7 was expressed in 10 (63%) of 16 pulmonary carcinoid tumors and only 5 (11%) of 46 GI carcinoid tumors (P <.001). Pancreatic endocrine tumors showed CK7 positivity in 6 (50%) of 12 cases, which was similar to the findings in lung carcinoids and significantly higher than in GI carcinoids (P <.01). CK20 was expressed in 0 (0%) of 16 pulmonary carcinoid tumors, in contrast to 24% and 33% of GI carcinoid tumors (P <.05) and PETs (P <.05), respectively. TTF-1 expression was highly specific for pulmonary carcinoid tumors. This peptide was present in 11 (69%) of 16 pulmonary carcinoid tumors and in only 1 (2%) of 46 and 0 (0%) of 12 GI carcinoid tumors (P <.001) and PETs (P <.001), respectively. A CK7(+)/CK20(-)/TTF-1(+) immunopanel result was moderately sensitive (sensitivity, 50%), and highly specific (specificity, 100%), for a diagnosis of pulmonary carcinoid tumor. CK7, CK20, and TTF-1 did not differ significantly between carcinoid tumors located in different sites of the GI tract. However, a trend was observed toward a lower prevalence of CK20 positivity in gastric tumors (P =.06) than in GI carcinoid tumors from the small intestine, colon, or appendix. Expression of CK7 and CK20, an

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoid Tumor; Diagnosis, Differential; Female; Gastrointestinal Neoplasms; Humans; Immunoenzyme Techniques; Insulinoma; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

Lung cancer is biologically and clinically classified as non-small-cell lung cancer (NSCLC) or small cell lung cancer (SCLC). NSCLC is accounting for about 80% of lung cancers. Serum tumour markers may be helpful in diagnostic of this cancer and in monitoring of the tumour growth or tumour volume reduction. Recent studies have focused on a new family of markers--hematopoietic cytokines, defined also hematopoietic growth factors (HGFs). It has been shown that the actions of HGFs are not limited to hematopoietic cells but can also affect the proliferation of nonhematopoietic cells. Some clinical investigations have shown cell surface receptors for macrophage--colony stimulating factor (M-CSF) in cancer cells and autologous production of M-CSF in various human cell lines derived from cancer. The purpose of this investigation was to compare serum levels of M-CSF in NSCLC patients to a control group, to assess pre- and post treatment levels of M-CSF in relation to levels of commonly accepted tumour markers such as carcinoembryonic antigen (CEA) and cytokeratin fragment 19 (CYFRA 21-1), and to define the diagnostic sensitivity of G-CSF in NSCLC. In this study, the serum levels of tumour markers were measured in 34 patients with NSCLC and in 20 healthy subjects. Serum samples were drawn before surgery and 10, 30, 90, 180 and 270 days after surgery. M-CSF and CEA were assayed using ELISA system and CYFRA 21-1 was measured by radioimmunoassay (RIA). The serum level of M-CSF was significantly increased in cancer patients relative to the control group on the 10th day after operation. Concentrations of CYFRA 21-1 were decreased on the 10th day, CEA on the 30th day and M-CSF on the 90th day after surgery. The diagnostic sensitivity of M-CSF was 55%, CEA--62% and CYFRA 21-1-51%. The diagnostic sensitivity and the serum level of M-CSF were related to the stage of NSCLC. These results suggest that M-CSF may be useful in diagnostic and monitoring of NSCLC, but it needs further studies.

Topics: Adult; Aged; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Lung Neoplasms; Macrophage Colony-Stimulating Factor; Male; Middle Aged; Neoplasm Staging

We describe the clinicopathological characteristics of 15 cases of primary signet ring cell adenocarcinoma of the lung and highlight the importance of recognizing that not all adenocarcinomas with signet ring cell features represent metastatic adenocarcinomas.. We evaluated the clinicopathological and immunohistochemical features of 15 cases of signet ring cell adenocarcinoma of the lung. The patients were 12 men and three women, age 30-75 years (mean 52.5 years). No evidence of a primary tumour elsewhere could be found on thorough clinical examination. Nine patients underwent resection and the remainder were biopsied. The tumours ranged from 18 to 80 mm in greatest dimension. Microscopically, two distinct patterns of growth were recognized: acinar and diffuse. The tumours were characterized by the presence of >75% signet ring cells. Periodic acid-Schiff and mucicarmine showed strong intracellular positive staining. Immunohistochemical stains for TTF-1 (6/6) and CEA (9/9) showed strong positive reaction in all cases evaluated. Three out of six cases were also positive for cytokeratin 7. All the tumours (6/6) were negative for cytokeratin 20, ER, PR and GCDFP-15. Follow-up information was obtained in 11 patients; six patients died within 1 year and five patients were alive from 3 to 36 months after initial diagnosis.. These cases highlight an unusual histological growth pattern of primary lung adenocarcinoma that may be mistaken for a metastasis from an occult primary. The recognition of this pattern of lung tumours is important for proper treatment.

Topics: Adult; Aged; Carcinoembryonic Antigen; Carcinoma, Signet Ring Cell; Female; Humans; Immunohistochemistry; Keratin-7; Keratins; Lung Neoplasms; Male; Middle Aged; Nuclear Proteins; Thyroid Nuclear Factor 1; Transcription Factors

We sought to determine the critical diameter of a peripheral non-small cell lung cancer tumor less than which no evidence of nodal micrometastasis is present.. Samples of 3081 lymph nodes from 181 patients with stage I peripheral lung cancer (155 with adenocarcinoma and 26 with squamous cell carcinoma) who had undergone complete resection with systematic lymphadenectomy were used in the study. In the samples immunohistochemical staining for cytokeratin was performed. The expression of vascular endothelial growth factor (VEGF) at primary sites was also immunohistochemically assessed.. Nodal micrometastasis was detected in 44 patients. The mean tumor sizes were 2.2 +/- 1.3 cm (range, 1.0-7.0 cm) in nodal micrometastasis-positive adenocarcinoma, 2.1 +/- 0.9 cm (range, 0.5-6.0 cm) in nodal micrometastasis-negative adenocarcinoma, 4.8 +/- 2.3 cm (range, 2.2-10.0 cm) in nodal micrometastasis-positive squamous cell carcinoma, and 3.2 +/- 2.1 cm (range, 0-9.0 cm) in nodal micrometastasis-negative squamous cell carcinoma. The tumor size in the nodal micrometastasis-positive group tended to be greater than that in the nodal micrometastasis-negative group in squamous cell carcinomas, but there was no significant difference in adenocarcinomas. Nodal micrometastasis was not found in patients with squamous cell carcinoma of 2.0 cm or less in diameter. However, nodal micrometastasis was found in 20% (19/95) of the patients with adenocarcinoma of 1.1 to 2.0 cm in diameter and even in 4 of 11 patients with adenocarcinoma of 1.0 cm or less. Among the patients with nodal micrometastasis, survival of patients with vascular endothelial growth factor overexpression was worse than that of patients without it. The survival of patients with nodal micrometastasis without vascular endothelial growth factor overexpression was comparable with that of patients without nodal micrometastasis.. A limited surgical intervention without lymphadenectomy is validated for squamous cell carcinoma of 2.0 cm or less without pleural involvement. In adenocarcinoma the tumor size itself is not a reliable guide for nodal micrometastasis status. In patients with nodal micrometastasis with vascular endothelial growth factor overexpression, the risk of systemic disease should be considered.

Topics: Adenocarcinoma; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Endothelial Growth Factors; Female; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Lymphokines; Male; Middle Aged; Protein Isoforms; Time Factors; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Clear cell (CRCC), papillary (PRCC) and chromophobe (CHRC) renal cell carcinoma (RCC) are the three most frequent subtypes of RCC. The rate and distribution of their metastatic lesions have not been well documented. We compared metastatic RCC according to subtype and primary tumor characteristics to better understand their behavior and to aid in the diagnosis of metastatic RCC. Pathology reports and clinical charts related to 283 CRCC, 48 PRCC and 13 CHRCC, including their respective sarcomatoid variants, were reviewed. A hundred and thirty-seven CRCC, 5 PRCC and 1 CHRCC with metastases were identified. CRCC and non-CRCC (PRCC and CHRCC) had different patterns of metastasis and primary tumor growth. CRCC metastases were predominantly distributed in lungs, bone, brain, lymph nodes, and adrenal glands. The associated primary CRCC measured 1.5 to 15 cm, were of all grades and stages, and were often associated with invasion of small or large veins. Three PRCC had regional lymph node metastases, 1 PRCC had both regional and mediastinal lymph node metastases. Bone metastasis was present in 1 case each of PRCC and CHRCC. One PRCC with metastasis solely to regional nodes measured 4 cm. The other 4 cases of PRCC with regional lymph node and/or distant metastases as well as the CHRCC with distant metastases were greater than 8 cm in diameter. In metastasizing and non-metastasizing non-CRCC, invasion of small veins was rare and invasion of renal veins was not seen. We cannot comment with any certainty on the metastatic behavior of CHRCC. In our experience, PRCC tend to loco-regional invasion with lymph node spread. They have a low potential for vascular invasion and distant metastases that likely occur only at late stages of the disease. CRCC has a propensity for vascular invasion and may be associated with distant metastasis at an early stage. Therefore, metastatic RCC at a distant location are most likely to be of CRCC origin than PRCC origin.

Topics: Adrenal Gland Neoplasms; Aged; Bone Neoplasms; Brain Neoplasms; Carcinoma, Renal Cell; Female; Humans; Immunoenzyme Techniques; Keratins; Kidney Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Survival Rate; Vimentin

The CYFRA 21-1 assay which detects the cytokeratin 19 (CK19) fragment is widely used as a tumor marker for lung cancer. We previously suggested that the failure of PCR amplification of exon 1 is closely related to the inability of the expression of mRNA for CK19, and hypothesized that point mutations might exist within exon 1. In order to prove this, sequence analysis of the promoter region and exon 1 was performed in 14 human lung cancer cell lines. Among the 14 lung cancer cell lines evaluated, point mutations within the promoter region (at -99, G-->C) of the CK19 gene were demonstrated in two cell lines (Lu135 and HI1017). In addition, point mutations within exon 1 (at 90, T-->C, Ala-->Ala and at 179, G-->C, Gly-->Ala) were also demonstrated in three cell lines (LU135, HI1017, and LC2/AD). Point mutations within the promoter region of CK19 (at -99) and within exon 1 (at 179) were confirmed by analysis of digestion by specific restriction enzymes. Since the same point mutation within exon 1 (at 179) was observed in genomes of normal volunteers, this mutation was considered as a single nucleotide polymorphism. In contrast, there were no mutations within the promoter region of exon 1 in genomes of normal volunteers. After a computer search, it was demonstrated that several transcription factors bind to the sense primer sequence which was designed for amplification of exon 1. In addition, after point mutations within the promoter region occurred (at -99), new sequences appeared to which known transcription factors (AP2) bind. In conclusion, analysis of genomic DNA for CK19 suggested that expression of mRNA for CK19 was regulated by several transcription factors which bound to the specific sequence with the promoter region of the CK19 gene. It was also suggested that the mutation in the promoter region of the CK19 gene down-regulated the expression of mRNA for CK19.

Topics: Antigens, Neoplasm; Exons; Humans; Keratin-19; Keratins; Lung Neoplasms; Point Mutation; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Sequence Analysis, DNA; Tumor Cells, Cultured

Signet-ring cell carcinoma (SRCC) of lung is a rare variant of pulmonary adenocarcinoma. In view of this rarity, the question of whether an SRCC is primary pulmonary or metastatic arises frequently because the majority of SRCCs seen in lung are metastatic tumors having arisen in stomach, colon, or breast. On routine histologic examination it is difficult to distinguish between pulmonary SRCC from SRCC metastasizing from other organs. Thyroid transcription factor-1 (TTF-1) is a homeodomain-containing transcription factor that is almost exclusively expressed in thyroid and pulmonary epithelial cells. TTF-1 expression has been demonstrated in various neoplasms of lung; however, the expression of TTF-1 in SRCCs has not been investigated so far. In the present study, using an immunoperoxidase staining procedure on paraffin sections, we investigated the expression of TTF-1, cytokeratin 7, cytokeratin 20, and villin (a specific marker expressed in tumors of the digestive tract, renal proximal tubules, and hepatic bile ducts) in 32 SRCCs from various organs (17 lung, 5 breast, 5 stomach, and 5 colon). Fourteen (82.4%) of 17 pulmonary SRCCs exhibited TTF-1 positivity, whereas none of the SRCCs of other organs were positive for TTF-1. A cytokeratin profile (CK7+/CK20-) was identified in 94.1% of pulmonary SRCC, and although it differed from the profile exhibited in colonic SRCCs (CK7-/CK20+), a similar profile was seen in breast SRCCs and some SRCCs arising in the stomach. Villin was identified in 29.4% of pulmonary SRCCs and 20% (one case) arising in the breast. Although the pattern of villin immunostaining exhibited by nondigestive tract SRCCs (cytoplasmic) differed from those of digestive tract SRCCs (membranous), distinguishing between the two groups based on their pattern of immunostaining alone would be difficult. The results of this study indicate that TTF-1 is expressed in a high percentage of pulmonary SRCCs and is very specific and that TTF-1 would be extremely valuable in distinguishing pulmonary SRCCs from those arising in other organs.

Topics: Biomarkers, Tumor; Carcinoma, Signet Ring Cell; Carrier Proteins; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Microfilament Proteins; Neoplasm Metastasis; Nuclear Proteins; Sensitivity and Specificity; Thyroid Nuclear Factor 1; Transcription Factors

The presence of non-tumor cells inside cancer tissue is one of the causes of errors in cell cycle analysis by DNA flow cytometry. The recent establishment of bivariate cytokeratin and DNA flow cytometry has made feasible the accurate assessment of tumor proliferative activity.. Bivariate flow cytometry and immunohistochemistry examinations of paraffin-embedded specimens were performed in 92 patients with non-small cell lung cancer (NSCLC). Determination of the S-phase fraction by flow cytometry, with cytokeratin gating (CK-gated SPF) and without gating (ungated SPF), and the expression of proliferating cell nuclear antigen by immunohistochemistry (PCNA labeling index), were used to assess cancer cell proliferation.. Two tumors had DNA histograms with a coefficient of variation of more than 8.0% and were excluded from the flow cytometric analysis. In DNA diploid tumors (n = 25), the ungated SPFs (8.7 +/- 3.6%) showed a lower distribution than the CK-gated SPFs (14.3 +/- 4.7%) (P < 0.0001). In DNA aneuploid tumors (n = 65), there was no difference in distribution between the ungated SPFs (15.0 +/- 8.3%) and the CK-gated SPFs (15.1 +/- 7.1%) (P = 0.94). The CK-gated SPF and the PCNA labeling index of an individual tumor had a good correlation (P < 0.0001), and this agreed with the result showing that DNA diploid and aneuploid tumors had equal proliferative activity (P = 0.64 and P = 0.63, respectively).. The technique using CK-gating markedly improved the SPF measurement in DNA diploid tumors. This assessment showed no difference in proliferative activity between DNA diploid and aneuploid tumors in NSCLC. Bivariate cytokeratin and DNA flow cytometry is an accurate and objective method for cancer-specific analysis, and will surely be informative in clinical oncology.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Carcinoma, Non-Small-Cell Lung; Cell Cycle; Cell Division; DNA, Neoplasm; Female; Flow Cytometry; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Paraffin Embedding; Proliferating Cell Nuclear Antigen

Topics: Adenocarcinoma; Aged; Aqueous Humor; Biomarkers, Tumor; Carcinoembryonic Antigen; Cytological Techniques; Eye Neoplasms; Humans; Immunoenzyme Techniques; Immunophenotyping; Keratins; Lung Neoplasms; Male; Syndrome; Tomography, X-Ray Computed; Uveitis; Vitreous Body

In small-cell lung carcinoma (SCLC) tumour cell contamination of leukaphereses is unknown. The present study was performed to define appropriate markers for reverse transcriptase polymerase chain reaction (RT-PCR), then to assess the contamination rate of leukaphereses and corresponding bone marrow samples. Immunocytochemistry (ICC) and RT-PCR methods were also compared. Among the 33 patients included, analyses were performed in 16 who had multiple leukaphereses and 17 who had only bone marrow. Leukapheresis products and bone marrow were analysed by ICC using several specific monoclonal antibodies against neural-cell adhesion molecule (N-CAM), epithelial glycoprotein (EGP-40) and cytokeratins (CK). Samples were also analyzed by RT-PCR for expression for N-CAM, synaptophysin, neuron-specific enolase, chromogranin, cytokeratin-18/-19, CEA, EGP-40, apomucin type 1 (MUC-1) and human endothelial cell-specific molecule (ESM-1). Using ICC staining, contaminating tumour cells were detected in 34% of leukaphereses (27% in patients with limited disease and 43% in those with extensive disease). N-CAM was the most reliable marker for detection of contamination. For RT-PCR, CK-19 and CEA were the only appropriate markers. Positive signal rate in leukaphereses increased to 78% (89% for patients with limited disease and 67% for extensive disease). In bone marrow, both techniques were in agreement whereas in leukaphereses, RT-PCR was better than ICC. A high rate of tumour cell contamination was demonstrated not only in bone marrow but also in leukaphereses from SCLC patients. The most appropriate technique was RT-PCR mainly in patients with limited disease.

Topics: Adult; Aged; Antigens, Neoplasm; Bone Marrow; Carcinoembryonic Antigen; Carcinoma, Small Cell; Cell Adhesion Molecules; Chromogranins; Epithelial Cell Adhesion Molecule; Female; Gastric Mucins; Humans; Immunohistochemistry; Keratins; Leukapheresis; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Neoplastic Cells, Circulating; Neural Cell Adhesion Molecules; Phosphopyruvate Hydratase; Proteins; Proteoglycans; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm; Synaptophysin

Immunohistochemistry is a useful method in the differential diagnosis between pleural mesotheliomas and metastatic adenocarcinomas to the pleura. Cytokeratin (CK) 5-6 is one of the most specific mesothelioma-associated antibodies. Cytokeratin 20 and CK7 have been used successfully in studies determining primary location of adenocarcinomas from metastases. In the current study, the value of these CKs in differential diagnosis of malignant pleural lesions was examined.. Ninety-three autopsy-verified cases (14 mesotheliomas and 79 adenocarcinomas including 42 primary lung tumors and 37 adenocarcinomas metastatic to the pleura) were stained on CK20, CK7, and CK5-6 with commercially available primary antibodies. The staining was conducted in an automated immunohistochemical system. The results were analyzed statistically at different positivity thresholds: 10% and 0%.. None of the mesotheliomas stained positively for CK20 at the 10% positivity level, but 3 cases showed focal positivity in < 10% of the tumor cells. Eighty-six percent (12 of 14) of these tumors were CK7+ and 64% (9 of 14) were CK5-6+. None of the mesotheliomas expressed the CK20+/7- pattern. Lung adenocarcinomas, both primary and metastatic, and breast carcinomas were very similar to mesotheliomas with regard to expression of CK20 and CK7 but differed significantly with regard to expression of CK5-6. Conversely, gastrointestinal adenocarcinomas and pancreaticobiliary tumors expressed CK20 positivity in a high proportion, 86% (13 of 15) and 77% (7 of 9), respectively. The gastrointestinal tumors stained positively for CK7 in only 20% (3 of 15) of cases and differed significantly from the other adenocarcinomas in this aspect. The CK20+/7- pattern was typical for gastrointestinal tumors.. Adding CK20 and CK7 to the panel of antibodies in the differential diagnosis of pleural mesothelioma versus metastatic adenocarcinomas is useful because diffuse CK20 positivity seems to be an indicator of metastasis. Furthermore, CK7 negativity most often is associated with metastases, and the CK20+/7- pattern, typical of colorectal adenocarcinomas, is absent in pleural mesotheliomas.

Topics: Adenocarcinoma; Autopsy; Biomarkers, Tumor; Diagnosis, Differential; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Mesothelioma; Neoplasm Metastasis; Pleural Neoplasms; Sensitivity and Specificity

Human Papillomavirus type 16 (HPV-16) is the cause of both benign lesions and ano-genital cancers. In HPV-associated cancers the transforming properties of the expressed viral E6 and E7 proteins have been revealed by a number of different assays. We have generated transgenic mice expressing HPV-16 E6/E7 genes under the control of the murine keratin 5 gene promoter, which should confer cell-type specific expression in the basal cells of squamous stratified epithelia. Transgenic mice developed thymic hyperplasia and lung neoplasia with 100% frequency, the thymus showing a size increase at 2 months and reaching the maximum dimension at 6 months, when lung carcinomas appeared. After this time the size of hyperplastic thymi decreased, while malignant formations invaded the mediastinal area. Hepatic metastasis could be also observed in some of the animals at the autopsy and death invariably occurred around 10-11 months of age.

Topics: Animals; Carcinoma; Keratin-15; Keratin-5; Keratins; Liver Neoplasms; Lung Neoplasms; Mice; Mice, Transgenic; Oncogene Proteins, Viral; Organ Size; Papillomavirus E7 Proteins; Papillomavirus Infections; Promoter Regions, Genetic; Recombinant Fusion Proteins; Repressor Proteins; Thymus Gland; Thymus Hyperplasia; Tumor Virus Infections

Topics: Biomarkers, Tumor; Female; Humans; Intestinal Perforation; Jejunal Neoplasms; Keratins; Lung Neoplasms; Middle Aged

High-grade invasive ductal carcinomas (IDCs) of the breast with large, central acellular zones on their cut surfaces are usually associated with the myoepithelial immunophenotype of carcinoma cells, which includes the expression of S-100 protein, alpha-smooth muscle actin, and keratin 14. To clarify the clinical significance of these features of IDCs, the authors compared the incidence of the myoepithelial immunophenotype immunohistochemically, patient prognosis, and metastatic sites of the tumor between 20 high-grade IDCs with large, central acellular zones and 40 control high-grade IDCs without these zones. The myoepithelial immunophenotype was detected in 16 IDCs (80%) with large, central acellular zones but in only seven IDCs (18%) without. The risk ratio of metastasis, especially in the brain and lung, and death from cancer were significantly higher (p = 0.0096 and p = 0.030) for the 20 IDCs with large, central acellular zones than for those without by Cox's univariate analysis. Using Cox's multivariate analysis, large, central acellular zones in IDCs were an indicator of high risk of brain and lung metastases and of death by cancer independent of nodal status and tumor size. Examination of large, central acellular zones and myoepithelial immunophenotype in high-grade IDCs appears helpful in predicting patient prognosis and preferential metastatic sites of the tumors.

Topics: Actins; Adult; Aged; Brain Neoplasms; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Differentiation; Female; Follow-Up Studies; Humans; Immunohistochemistry; Immunophenotyping; Keratins; Lung Neoplasms; Middle Aged; Myoepithelioma; Prognosis; S100 Proteins; Survival Rate

The T1, N0, M0 subset of stage I lung adenocarcinoma is a tumor that has a 5-year disease-free survival rate of 66% to 85%. To date, there has not been a rigorous immunohistochemically detected lymph node micrometastasis study composed of patients with identical stage and type of tumors, and in which standard histologic features were incorporated into multivariate analyses. We immunohistochemically examined the peribronchial and mediastinal lymph nodes from 80 consecutively accrued patients with T1, N0, M0 adenocarcinomas and bronchioloalveolar carcinomas unselected for distant metastasis, and an additional 39 patients with similar stage and type neoplasms who were selected for their development of metastases to evaluate the prevalence of micrometastases, their association with distant metastases, and their relationship with other pathologic prognostic features. All slides were stained with keratin AE1/3. Micrometastases were confirmed with Ber-Ep4. Three immunohistochemically detected lymph node micrometastases were identified in three of 80 consecutively accrued patients (4%). These three positive stains constituted 0.5% of the 573 stains required to immunohistochemically screen all of the lymph node blocks from these patients. Among the 39 patients who were selected because they developed distant metastases, three immunohistochemically detected lymph node micrometastases from three patients were identified, which constituted 8% of patients in this group and 1% of the 280 stains required to screen all of these patients' lymph nodes. Small vessel invasion, maximum tumor dimension, and immunohistochemically detected lymph node micrometastases were independently associated with metastases on multivariate analysis. Among patients who developed metastases, there was no significant difference in the disease-free survival rate between those with and those without immunohistochemically detected lymph node micrometastases. Given the low sensitivity in terms of the number of immunohistochemical stains performed, and the prognostic significance of standard histologic features, the use of immunohistochemical screening lymph nodes from all patients with T1, N0, M0 adenocarcinomas is questionable.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Antigens, Surface; Biomarkers, Tumor; Bronchi; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Mediastinum; Neoplasm Staging; Proportional Hazards Models

Anti-CEA, anti-vimentin, CAM5.2, BerEp4, Leu-M1 and anti-EMA were applied to effusions from 36 mesotheliomas, 53 adenocarcinomas and 24 reactive mesothelial proliferations. Stepwise logistic regression analysis selected three criteria of major importance for distinguishing between adenocarcinoma and mesothelioma: BerEp4, CEA and EMA accentuated at the cell membrane (mEMA), these three being of similar diagnostic value. The pattern BerEp4-, CEA- and mEMA+ was fully predictive for mesothelioma (sensitivity 47%), whereas the opposite pattern was fully predictive for adenocarcinoma (sensitivity 80%). Only EMA seemed to distinguish between mesotheliosis and mesothelioma. Comparison of reactivity in cytological and histological material from the same mesotheliomas showed similar staining frequencies for CEA and CAM5.2, with some random variation for Leu-M1 and EMA, whereas vimentin and BerEp4 reactivity was more frequent in cytological specimens.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antibodies, Neoplasm; Antibody Specificity; Antigens, Neoplasm; Antigens, Surface; Biomarkers; Biomarkers, Tumor; Carcinoembryonic Antigen; Diagnosis, Differential; Epithelium; Humans; Hyperplasia; Immunoenzyme Techniques; Keratins; Lewis X Antigen; Logistic Models; Lung Neoplasms; Mesothelioma; Mucin-1; Neoplasm Proteins; Pleural Effusion, Malignant; Sensitivity and Specificity; Vimentin

We examined differences in the 2-year survival rate in bronchogenic squamous cell carcinoma patients with normal serum levels of both cytokeratin 19 fragment (CYFRA 21-1) and squamous cell carcinoma-related antigen (SCC) (NL group, n=15), patients with only increased SCC levels (SCC+ group, n=14), patients with only increased CYFRA 21-1 levels (CYFRA+ group, n=14), and patients with elevated levels of both CYFRA 21-1 and SCC (EL group, n=65). The 2-year survival rates for the CYFRA+ and the EL group were lower than those for the SCC+ group and the NL group (0 and 12.9% vs. 66.7 and 85.6%, log rank: p<0.0001, Wilcoxon: p<0.0001). However, there were no differences between the rate for the SCC+ and the NL group and between that for the CYFRA+ and the EL group. Serum carcinoembryonic antigen (CEA) levels increased in the patients with the elevated CYFRA 21-1 levels. This results suggest that there may be some differences between squamous cell carcinoma patients with increased CYFRA 21-1 levels and those with normal levels and that it is CYFRA 21-1 levels, not SCC levels, that relate to the prognosis.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Carcinoma, Squamous Cell; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Probability; Prognosis; Survival Rate; Time Factors

We examined a microdissemination of cancer cells in lymph nodes and assessed its clinical and biologic characteristics.. Both primary tumors and lymph nodes (2030 nodes) were obtained from 122 patients with primary stage I lung cancer who underwent curative operations with routine systematic nodal dissection of both the hilar and the mediastinal nodes. Immunohistochemical anticytokeratin staining was used to detect nodal microdissemination of cancer cells. Vascular endothelial growth factor, vascular endothelial growth factor type C, and nm23 expression at primary sites were also immunohistochemically studied.. In total, 35 patients (29%) had cytokeratin-positive cells in lymph nodes. Increased expression of vascular endothelial growth factor (P =.0001) and vascular endothelial growth factor type C (P <. 0001) at primary sites were significantly associated with nodal microdissemination, and nm23 was inversely correlated with microdissemination (P =.008). The 3- and 5-year survivals for the patients with nodal microdissemination were 57% and 54%, respectively, which was a significantly worse prognosis as compared with those prognoses (83% and 76%) for the patients without nodal microdissemination (P =.006). The independent prognostic impact of nodal microdissemination was not clear; however, vascular endothelial growth factor retained independent significance.. All of these findings lead us to conclude that the microspread of tumor cells in nodes detected by immunohistochemical anticytokeratin staining is definitely a metastasis with a high risk of systemic disease.

Topics: Aged; Aged, 80 and over; Analysis of Variance; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Endothelial Growth Factors; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Lymphokines; Male; Middle Aged; Monomeric GTP-Binding Proteins; NM23 Nucleoside Diphosphate Kinases; Nucleoside-Diphosphate Kinase; Prognosis; Survival Rate; Transcription Factors; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factors

The epidermal growth factor receptor (EGFR) is overexpressed in 50-70% of human primary breast, lung, and colon carcinomas, whereas it is not usually expressed in hematopoietic cells. We developed a novel reverse transcription-PCR (RT-PCR)-Southern blot assay for the detection of circulating, EGFR mRNA-expressing tumor cells in carcinoma patients. The assay was set up by increasing the amount of cDNA step by step in the PCR reaction. The highest sensitivity and specificity were found when using 800 ng of cDNA in the PCR reaction. Peripheral blood samples from 91 patients with either colon (38), lung (30), or breast (23) carcinomas and from 38 healthy volunteers were analyzed. EGFR transcripts were found in 44 of 75 (59%) patients with metastatic carcinoma and in 4 of 38 (10.5%) healthy donors (P < 0.001; chi2 test). The expression of EGFR, cytokeratin 19, and carcinoembryonic antigen mRNA in blood samples from patients with metastatic colon carcinoma was compared. EGFR, cytokeratin 19, and carcinoembryonic antigen transcripts were found in 8 of 11 (73%), 3 of 11 (27%), and 5 of 11 (45%) patients, respectively. Furthermore, two of seven (29%) Dukes' B and five of nine (55%) Dukes' C colon carcinoma patients were found to express EGFR mRNA in the peripheral blood. All patients that expressed EGFR transcripts in the peripheral blood were found to express the EGFR protein in the corresponding primary carcinoma, as assessed by immunohistochemistry. These data suggest that the EGFR assay that we developed is a highly specific and sensitive technique to detect circulating tumor cells in patients affected by different carcinoma types.

Topics: Breast Neoplasms; Carcinoembryonic Antigen; Colonic Neoplasms; ErbB Receptors; Female; Gene Expression Regulation, Neoplastic; Humans; Keratins; Lung Neoplasms; Neoplasm Staging; Neoplasms; Neoplastic Cells, Circulating; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Transcription, Genetic; Tumor Cells, Cultured

Metastases to the oral mucosa are rare, representing less than 1% of the tumors at this site. Most of these metastatic neoplasms originate in the lungs, kidneys, and liver.. The clinicopathologic features of an occult hepatocellular carcinoma, metastatic to the oral mucosa, are reported. The patient, a 70-year-old male, complained of 3 distinct polypoid, reddish lesions of the antero-inferior alveolar crest and both the right and left postero-superior attached gingiva, without bone involvement. The lesions were excised, with the clinical diagnosis of multiple vascular tumors, formalin-fixed, paraffin-embedded, cut and stained with hematoxylin and eosin. Consecutive sections were immunostained for alpha-1-antichymotrypsin, CEA, cytokeratins, EMA, hepatocyte antigen, PSA, S-100 protein, and thyroglobulin, using the alkaline phosphatase/anti-alkaline phosphatase technique.. The morphologic features of the lesions were consistent with the diagnosis of carcinoma with trabecular and glandular patterns and bile secretion; furthermore, immunohistochemical reactivity for alpha-1-antichymotrypsin, cytokeratins, CEA, EMA, and hepatocyte antigen was demonstrated and the hepatic origin of the tumor was postulated. Ultrasonography demonstrated a liver mass, which was biopsied and treated by chemoembolization. While no further complications occurred in the oral mucosa, the patient died 8 months after the diagnosis for widespread diffusion of the tumor to the lungs and brain.. This case emphasizes the need to include metastatic tumors in the differential diagnosis of atypical neoplasms of the oral mucosa and to evaluate the opportunity of surgical treatment in order to preserve the functions of the mouth, even if the prognosis of the primary tumors remains unfavorable.

Topics: Aged; alpha 1-Antichymotrypsin; Biomarkers, Tumor; Brain Neoplasms; Carcinoembryonic Antigen; Carcinoma, Hepatocellular; Chemoembolization, Therapeutic; Diagnosis, Differential; Fatal Outcome; Gingival Neoplasms; Humans; Keratins; Liver Neoplasms; Lung Neoplasms; Male; Mucin-1; Polyps; Vascular Neoplasms

Pulmonary large-cell neuroendocrine carcinoma (LCNEC) is a newly proposed clinicopathologic entity; a few cases of LCNEC have been reported in other sites, such as the uterine cervix and the thymus. In the salivary glands, LCNEC is extremely rare and is not recognized as a specific entity in the World Health Organization classification. We retrospectively reviewed from our files 1675 cases of surgically resected primary parotid gland tumors and found 2 cases of LCNEC that fulfilled the criteria of pulmonary LCNEC. These cases occurred in 72- and 73-year-old men who had short histories of enlarging parotid gland tumors. The tumors were composed of large cells that exhibited organoid, solid, trabecular, and rosette-like growth patterns with a high mitotic rate and a conspicuous tendency for necrosis. The tumor cells were polygonal and characterized by a moderate nuclear:cytoplasmic ratio, coarse chromatin, and conspicuous nucleoli. Immunohistochemical examination revealed that the tumor cells were positive for six general neuroendocrine markers, cytokeratin, p53, bcl-2, epidermal growth factor receptor, and cyclin D1. Markedly reduced expressions of p21Waf1 and p27Kip1 were also noticed. The Ki-67 labeling index was more than 50% in both cases. One case showed loss of heterozygosity at TP53 accompanied by a p53 gene point mutation. Loss of heterozygosity at chromosome 9p21 was detected in both cases; one was accompanied by a p16 gene silent point mutation. Both patients died of the disease, with recurrence 5 months and 4 years after surgery, respectively. These findings indicate that LCNEC is a rare but distinct salivary gland tumor with highly aggressive biologic behavior. Multiple alterations of cell cycle regulators and tumor suppressor genes may play an important role in presenting the biologic characteristics of this rare parotid gland tumor.

Topics: Aged; Base Sequence; Carcinoma, Large Cell; Carcinoma, Neuroendocrine; Carcinoma, Non-Small-Cell Lung; Cyclin D1; Diagnosis, Differential; DNA Mutational Analysis; DNA, Neoplasm; ErbB Receptors; Humans; Keratins; Ki-67 Antigen; Loss of Heterozygosity; Lung Neoplasms; Male; Microscopy, Electron; Parotid Neoplasms; Point Mutation; Proto-Oncogene Proteins c-bcl-2; Tumor Suppressor Protein p53

Serum pyridinoline cross-linked carboxy-terminal telopeptide of type I collagen (ICTP) is a metabolite of type I collagen comprising 90% or more of organic substances in bone. Its usefulness as a marker of bone metastasis from malignant tumors is expected.. We measured ICTP to evaluate its clinical usefulness for diagnosis of bone metastasis in 140 patients with lung cancer. For comparison, serum carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA 21-1), gastrin-releasing peptide precursor (ProGRP), alkaline phosphatase and calcium were simultaneously measured. ICTP was measured by double-antibody radioimmunoassay.. ICTP was significantly higher in patients with bone metastasis from lung cancer than in the group without bone metastasis, patients with other pulmonary diseases or healthy control subjects and showed excellent sensitivity and specificity, indicating that this marker is highly useful for complementary diagnosis of bone metastasis from lung cancer. Moreover, the survival duration was significantly shorter in the ICTP-positive group than in the ICTP-negative group, suggesting that ICTP can be a prognostic factor in lung cancer.. It is suggested that measurement of ICTP is worthwhile as a serological diagnostic method of bone metastasis from lung cancer. Moreover, since repeated measurements are possible, this measure was considered very helpful in complementary diagnosis of bone metastasis and also as a standard to determine the timing of examinations such as bone scintigraphy.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Alkaline Phosphatase; Antigens, Neoplasm; Biomarkers; Biomarkers, Tumor; Bone Neoplasms; Calcium; Carcinoembryonic Antigen; Carcinoma, Large Cell; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Collagen; Collagen Type I; Female; Gastrointestinal Hormones; Humans; Keratin-19; Keratins; Lung; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Peptides; Prognosis; Recombinant Proteins; Sensitivity and Specificity; Survival Rate

Sixty patients with squamous cell carcinoma (SCC) of the lung, including 25 cases with recurrence and 35 cases without recurrence 1 year after operation, were enrolled in this study. The serial serum levels of cytokeratin fragment 19 (CYFRA 21-1) and SCC antigen were measured before operation and 1 week, 1 month, 3 months, 6 months, 9 months, and 12 months after operation for early detection of recurrence. The results revealed that 1) mean serum values of CYFRA 21-1 were significantly higher at early and any times after operation in 25 patients with recurrent SCC when compared with 35 patients without recurrent SCC; and 2) mean serum values of SCC antigen were significantly higher until 9 and 12 months after operation, in 25 patients with recurrent SCC when compared with 35 patients without recurrent SCC. We conclude that CYFRA 21-1 is a better marker than SCC antigen for early prediction of SCC recurrence in the lung.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Immunoradiometric Assay; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Postoperative Period; Serpins

Small Cell Lung Cancer (SCLC), a clinically aggressive cancer, accounts for approximately 25% of primary lung cancers. We carried out suppression subtractive hybridization (SSH), a PCR-based method for cDNA subtraction, between the human classic, NCI-H69 and variant, more aggressive NCI-N417 SCLC cell lines to isolate and characterize variable expression of genes, which may be responsible for differential degree of tumorigenicity of SCLC. Using NCI-N417 as a tester, we obtained 28 differentially expressed cDNA clones from a total of 60 arbitrarily picked clones. Among the 28 cDNA clones, 4 were unknown genes, 2 were fatty acid binding protein (FABP) with specific identification of mRNA for mammary-derived growth inhibitor (MDGI), 1 was human alpha-enolase, 4 were ribosomal proteins, 2 were structural genes, vimentin and moesin (membrane-organizing extension spike protein), and 9 were homologous with murine leukemia viruses, whereas 2 others had enhanced expression in NCI-H69 and A549 cell lines, and 4 were cell surface proteins and murine type C retrovirus. Expression of FABP/MDGI was significantly high in NCI-H417, which may influence mitosis and cell growth as implicated in other tissues, contrary to the conclusion drawn for the role of MDGI in human breast cancer. Higher expression of ribosomal proteins in NCI-N417 compared to NCI-H69 may have a role in differential tumorigenicity and metastatic ability. Further, we obtained 14 differentially expressed cDNA clones by reversing the tester and driver, using NCI-H69 as a tester. Of these 14 differential cDNAs, 5 were unknown genes, 2 were specific for keratins, others had similarities with protease inhibitor, human BAC clone, Alu RNA binding protein, and tumor expression-enhanced gene. Characterization of these differentially expressed cDNA clones will provide useful information in understanding of the genes responsible for differential tumorigenicity of SCLC.

Topics: Base Sequence; Blotting, Northern; Carcinoma, Small Cell; Carrier Proteins; DNA, Complementary; Fatty Acid Binding Protein 3; Fatty Acid-Binding Protein 7; Fatty Acid-Binding Proteins; Gene Expression; Glutathione Transferase; Humans; Keratins; Lung Neoplasms; Molecular Sequence Data; Myelin P2 Protein; Neoplasm Proteins; Ribosomal Proteins; Tumor Cells, Cultured; Tumor Suppressor Proteins; Vimentin

To undertake a comparative evaluation of three antimesothelial markers (thrombomodulin, cytokeratin 5/6 and calretinin) with broad spectrum cytokeratin (AE1/AE3) in differentiating between sarcomatoid mesothelioma and a spectrum of spindle cell neoplasms.. Thirty-one malignant sarcomatoid mesotheliomas were studied. Calretinin expression was focally identified in 12 (39%) tumours and thrombomodulin and cytokeratin 5/6 immunoreactivity was seen in nine (29%) cases. In comparison there was strong diffuse cytoplasmic reactivity with the broad spectrum cytokeratin (AE1/AE3) in 24 of 31 (77%) tumours. Thirty mixed spindle cells neoplasms were studied. No calretinin expression was identified in any case. Thrombomodulin immunoreactivity was identified in four (16%) cases (two angiosarcomas, two high-grade sarcomas, not otherwise specified). Cytokeratin 5/6 expression was seen in one high-grade pulmonary sarcoma originally termed malignant fibrous histiocytoma. None of the antimesothelial markers was expressed in the four spindle cell carcinomas studied. In contrast, broad spectrum cytokeratin was diffusely expressed in all four spindle cell carcinomas (three pulmonary, one renal), both synovial sarcomas, both malignant mixed Müllerian tumours, one of three pulmonary leiomyosarcomas and two of nine sarcomas, not otherwise specified.. Immunohistochemistry has a more limited role in the diagnosis and distinction of sarcomatoid mesothelioma from other spindle cell neoplasms. The combination of a broad spectrum cytokeratin with calretinin combines both high sensitivity (77% for AE1/AE3) with high specificity (100% for calretinin) for sarcomatoid mesothelioma and can be diagnostically useful. The mesothelial markers, thrombomodulin and cytokeratin 5/6, are not useful alone in the diagnosis of sarcomatoid mesothelioma as each shows insufficient antibody sensitivity, although together they complement calretinin.

Topics: Biomarkers, Tumor; Calbindin 2; Carcinoma; Carcinoma, Renal Cell; Carcinoma, Squamous Cell; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Lung Neoplasms; Mesothelioma; S100 Calcium Binding Protein G; Sarcoma; Thrombomodulin

Transitional cell carcinoma (TCC), a neoplasm of urinary bladder urothelial cells, generally appears in either of two forms, papillary non-invasive or invasive TCC, although intermediate forms can occur. Each has a distinctive morphology and clinical course. Altered expression of the p53 and pRb genes has been associated with the more serious invasive TCC, suggesting that the loss of activity of these tumor suppressor proteins may have a causal role in this disease. To test this hypothesis directly, transgenic mice were developed that expressed the simian virus 40 large T antigen (TAg) in urothelial cells under the control of the cytokeratin 19 gene (CK19) regulatory elements. In one CK19-TAg lineage, all transgenic mice developed highly invasive bladder neoplasms that resembled invasive human bladder TCCs. Stages of disease progression included development of carcinoma in situ, stromal invasion, muscle invasion, rapid growth, and, in 20% of affected mice, intravascular lung metastasis. Papillary lesions never were observed. Western blot analysis indicated that TAg was bound to both p53 and pRb, which has been shown to cause inactivation of these proteins. Our findings support suggestions that (i) inactivation of p53 and/or pRb constitutes a causal step in the etiology of invasive TCC, (ii) papillary and invasive TCC may have different molecular causes, and (iii) carcinoma in situ can represent an early stage in the progression to invasive TCC.

Topics: Alkaline Phosphatase; Animals; Antigens, Polyomavirus Transforming; Blotting, Western; Carcinoma in Situ; Carcinoma, Transitional Cell; Cell Lineage; Disease Models, Animal; Disease Progression; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mice; Mice, Transgenic; Neoplasm Invasiveness; Neoplasm Transplantation; Precancerous Conditions; Retinoblastoma Protein; Transplantation, Heterologous; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

The histologic classification of pulmonary neoplasms can have important implications regarding appropriate management of patients. Although the histologic classification of lung tumors is predominantly based on morphology, ancillary studies such as immunohistochemistry can be used in difficult cases, and the diagnosis of large cell neuroendocrine carcinoma requires confirmation of neuroendocrine differentiation by immunohistochemistry or electron microscopy. We immunostained 142 lung tumors for B72.3, keratin 34betaE12, keratin 7, keratin 14, keratin 17, synaptophysin, and chromogranin to determine the utility of neuroendocrine markers and epithelial markers in the differential diagnosis. Among neuroendocrine carcinomas (small cell carcinoma and large cell neuroendocrine carcinoma), 84% (37 of 44) were chromogranin positive, 64% (21 of 36 small cell, 6 of 6 large cell neuroendocrine) were synaptophysin positive, 5% (2 of 43) were keratin 34betaE12 positive, 9% (4 of 44) were keratin 7 positive, and 5% (2 of 37) of small cell carcinomas and 50% (3 of 6) of large cell neuroendocrine carcinomas were B72.3 positive. Among non-neuroendocrine carcinomas, 5% (5 of 98) were chromogranin positive, 3% (3 of 96) were synaptophysin positive, and 97% (95 of 98) were positive for either keratin 34betaE12 or keratin 7 and 99% (97 of 98) were positive for either keratin 34betaE12, keratin 7 or B72.3. An antibody panel consisting of keratin 7, keratin 34betaE12, chromogranin, and synaptophysin separated 132 of 141 tumors (94%) into distinct groups. We conclude that immunostaining with both neuroendocrine markers and epithelial markers can be useful in the differential diagnosis of lung neoplasms.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Biomarkers, Tumor; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Chromogranins; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Pilot Projects; Synaptophysin

Two cases of bilateral radiation pneumonitis associated with unilateral thoracic irradiation against lung cancer are described. Both patients died of respiratory failure and autopsy was performed. Histologically, bilateral diffuse alveolar damage was demonstrated in both cases, associated with marked organization of hyaline membrane in one case (case 1). In addition, numerous hyperplastic type II pneumocytes which strongly expressed cytokeratins 8, 18 and 19 were observed. In both patients' sera, antibodies against cytokeratin 8, 18 and 19 were demonstrated by a Western immunoblot. The possible association between autoantibodies to cytokeratins and diffuse alveolar damage observed in patients with bilateral radiation pneumonitis are discussed.

Topics: Adenocarcinoma; Aged; Autoantibodies; Blotting, Western; Fatal Outcome; Humans; Keratins; Lung Neoplasms; Male; Pulmonary Alveoli; Radiation Pneumonitis; Sarcoma, Small Cell

The CYFRA 21-1 assay which detects cytokeratin 19 (CK19) fragment is widely used as a tumor marker for lung cancer. However, the reason why some lung cancer cell lines release CK19 fragment in culture supernatants and others do not, remains unclear. It was hypothesized that the release of CK19 fragment may be elucidated by the expression of mRNA for CK19. In order to prove this, the mRNA for CK19 was quantitatively evaluated by the competitive reverse transcriptase-polymerase chain reaction (competitive RT-PCR). The level of CYFRA 21-1 in the culture supernatant was measured by an immunoradiometric assay. CK19 protein synthesis was evaluated by a Western blotting and immunohistochemistry. Fourteen lung cancer cell lines were evaluated, and the amount of mRNA correlated well with the level of CYFRA 21-1 in culture supernatants. Analysis of genomic DNA for CK19 demonstrated that three cell lines which could not produce CYFRA 21-1, conjectured that some abnormalities in exon 1 or the 5'-region upstream from exon 1. In conclusion, it was demonstrated that the release of CK19 fragment was closely related to the expression of mRNA for CK19, and the possibility that genomic change of CK19 DNA down-regulated the expression of mRNA for CK19 was suggested.

Topics: Antigens, Neoplasm; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; DNA, Neoplasm; Down-Regulation; Gene Expression Profiling; Humans; Immunohistochemistry; Keratin-19; Keratins; Lung Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured

The aim of the study was to assess diagnosis value of tumor markers for differential diagnosis between mesothelioma and other pleural tumors.. Prospective study of 85 patients attending our hospital with malignant pleural effusion. The diagnostic approach involved routine pleurocentesis followed by pleural needle. When precise diagnosis was not achieved, thoracoscopy with pleural biopsies was performed. Carcinoembryonic antigen (CEA), hyaluronic acid, tissue polypeptide antigen and cyfra 21 to 1 were measured in serum and pleural fluid.. By using receiver operating characteristics curves and area under curves, the best diagnostic characteristics were obtained with pleural and serum CEA concentrations. The area under the curve was larger for pleural ACE than for serum ACE. The sensitivity and specificity of a pleural CEA level exceeding 3 ng/mL for ruling out the diagnosis of mesothelioma were 100% and 77%, respectively.. A CEA level above 3 ng/mL in pleural fluid eliminated the diagnosis of mesothelioma, whereas the other markers were not sufficiently discriminant. However, despite a negative predictive value of 100% at a cutoff of 3 ng/mL, CEA assay in pleural fluid only avoids a small number of diagnostic thoracoscopies.

Topics: Adenocarcinoma; Aged; Antigens, Neoplasm; Area Under Curve; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Humans; Hyaluronic Acid; Keratin-19; Keratins; Lung Neoplasms; Male; Mesothelioma; Middle Aged; Pleural Effusion; Pleural Neoplasms; Prospective Studies; ROC Curve; Sensitivity and Specificity; Tissue Polypeptide Antigen

A unique case of eccrine porocarcinoma with pulmonary lymphangitis and pericardial involvement is reported. The clinical course was aggressive, leading to the death of the patient a few months after diagnosis. Certain pathologial markers of clinical aggressiveness were retrospectively investigated: p53 and Ki-67 expression were determined by means of immunohistochemistry. Angiogenesis was assessed by determination of intratumor microvessel density at the vascular 'hot spot' with the anti-CD34 monoclonal antibody and quantitative analysis using computerized image analyzer. Both primary tumor and metastatic lymph node presented immunostaining for p53 and Ki-67, with a higher degree of vascularization in the secondary lesions compared to the primary tumor. Our findings suggest a correlation between tumor vascularization and clinicopathological parameters of aggressiveness in malignant eccrine porocarcinoma. Taking into account the disappointing results of current treatments for metastatic eccrine porocarcinoma, the assay of microvessel density may be helpful in selecting the patients of high risk for recurrence or death who may benefit of anti-angiogenic therapies.

Topics: Acrospiroma; Biomarkers, Tumor; Heart Neoplasms; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Lung Neoplasms; Lymphangitis; Lymphatic Metastasis; Male; Middle Aged; Neovascularization, Pathologic; Pericardial Effusion; Sweat Gland Neoplasms; Tumor Suppressor Protein p53

The purpose of the study was to evaluate the performance of dual-head coincidence gamma camera imaging using FDG in association with serum marker assays in identifying lung carcinoma in patients with abnormal findings on chest radiography.. A prospective evaluation of FDG imaging with coincidence detection emission tomography (CDET) using a dual-head gamma camera combined with the assessment of 3 sensitive serum markers of lung cancer (carcinoembryonic antigen, neuron specific enolase, and CYFRA 21-1) was performed on the same day on 58 consecutive patients with known or suspected lung malignancy.. Fifty-three patients were proven to have lung cancer, and 5 patients had benign lung disease. Coincidence imaging showed significantly increased FDG uptake in 49 of 53 patients with proven malignancy (sensitivity, 92.5%) and in 3 patients with benign disease. FDG imaging had negative findings in 4 patients with proven malignancy and 2 patients with benign disease. Serum tumor marker levels were elevated in 42 of 53 cancer patients (sensitivity, 79.2%) and normal in 11 patients with proven malignancy. Nine patients with proven malignancy had positive findings on FDG images and negative marker assays. Two patients with proven malignancy had negative findings on FDG images and positive marker assays. The positive predictive value for lung cancer was 94.2% for FDG alone and 97.6% for FDG in association with serum markers.. In this study, FDG CDET imaging was a powerful tool for evaluating patients with lung lesions suggestive of malignancy. Although the determination of serum marker levels was less accurate than FDG imaging, positive FDG results found in association with positive markers significantly increased the likelihood of lung malignancy.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Female; Fluorodeoxyglucose F18; Gamma Cameras; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Prospective Studies; Radionuclide Imaging; Radiopharmaceuticals; Sensitivity and Specificity

The predictive value of lymph node micrometastasis, detected by immunohistochemical or genetic methods, is well appreciated in terms of prognosis. However, a major problem is high false-positive rates, because most methods focus on cytokeratin, which is a component not only of carcinoma but also normal epithelial and nonepithelial cells. Mutant allele-specific amplification (MASA) can detect DNAs derived from cancer cells itself, reportedly with high sensitivity. It was, therefore, used with nested-PCR using p53 or K-ras mutation for analysis of lymph node micrometastasis in non-small cell lung carcinoma (NSCLC) patients in the present study, in comparison with the immunohistochemical method using an anti-cytokeratin reagent for the same samples. Lymph nodes from 31 NSCLC patients with p53 and K-ras mutated tumors (30 and 1, respectively) staged as pathological (p)-T1-4 N0-1 and M0 were examined. Genetic and immunohistochemical methods demonstrated positive reactions in 34 (15%) and 61 (27%) of 229 lymph nodes, respectively (9 cases, 29%, and 24 cases, 77%). The concordance with the two methods was 77%, but 13 (39%) of 34 genetically positive lymph nodes could not be detected by immunohistochemistry (IHC). Of 22 cases with p-N0 disease, 6 (27%) were genetically positive in hilar and/or mediastinal lymph nodes, and 4 (67%) of them died after cancer relapse. In contrast, none of the patients without micrometastasis died of cancer (P < 0.001, log rank analysis). Of the same p-N0 patients, 17 (77%) were positive by IHC, and 4 (24%) of them died of cancer, whereas 5 negative patients did not suffer cancer relapse. Survival did not significantly differ between cases positive and negative (P = 0.246) by IHC. According to the g-N (N factor restaged by a genetic method), patients with g-N1 and g-N2 disease had a shorter survival than those with g-N0 disease (P = 0.042 and P < 0.001, respectively). However, no significant difference was observed with grading by IHC. Thus, detection of micrometastasis in regional lymph nodes with the MASA method, in other words with a carcinoma-specific marker, is of greater prognostic significance for early stage NSCLC patients than immunohistochemical results. This approach should facilitate selection of patients for whom postoperative adjuvant chemotherapy should be performed.

Topics: Adenocarcinoma; Adult; Aged; Alleles; Carcinoma, Squamous Cell; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Mutation; Nucleic Acid Amplification Techniques; Polymerase Chain Reaction; Prognosis; Survival Analysis

Cell lysis and tumor necrosis release cytokeratin, a tumor marker of lung cancer, into the serum. The serum cytokeratin level can also be elevated in benign cavitary lung diseases. The purpose of this study was to evaluate whether Cyfra 21-1 can differentiate malignant lung diseases from benign diseases with cavitary lesions.. This study is a retrospective review of the case records of patients with lung lesions seen during a 4-year period from January 1993 to May 1996.. Serum Cyfra 21-1 levels were measured in 306 patients with lung cancer (n = 143) or benign lung disease (n = 163). The patients were grouped according to radiologic evidence of cavitary lung lesions. Lung cancer included both non-small cell (n = 123) and small cell (n = 20) lung cancers, and the benign diseases include tuberculosis (n = 87), abscess (n = 26), pneumonia (n = 4), and others (n = 46).. Although Cyfra 21-1 clearly differentiated cavitary lung cancer (15.0, 9.1-29.8 ng/ml, median and interquartile range, n = 39) from benign cavitary disease (P < 0.001), and noncavitary lung cancer from benign noncavitary disease (1.7, 0.9-2.6 ng/ml, n = 108, P < 0.001), it could not differentiate noncavitary lung cancer (5.0, 2.1-12.4 ng/ml, n = 104) from benign cavitary diseases (3.3, 1.4-8.3 ng/ml, n = 55, P = 0.45).. Serum Cyfra 21-1 is a useful tumor marker for differentiating benign from malignant lung diseases. However, different cutoff values are needed, depending on the presence of cavitary lesions. We recommend cutoff values of 30 ng/ml for cavitary lung diseases and 6 ng/ml for noncavitary lung diseases. If there are no radiologic data, a cutoff value of 15 ng/ml is recommended.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Reference Values; Retrospective Studies; Sensitivity and Specificity

Reverse transcriptase polymerase chain reaction (RT-PCR) is often used for sensitive detection of micrometastasis in peripheral blood, lymph nodes and bone marrow. While the utility of this method has been documented, it also has limitations in the detection of micrometastasis. The mRNA of target genes can be detected in healthy donors or in samples used for negative control, therefore the non-quantitativeness of conventional RT-PCR has been called into question. We analyzed the expression level of cytokeratin (CK) 18 mRNA in established esophageal and gastrointestinal carcinoma cell lines and non-epithelial cells, using quantitative RT-PCR, based on real time 'TaqMan TM' technology. CK 18 mRNA is more highly expressed in carcinoma cells than in non-epithelial cells. However, the expression level in non-epithelial cells was easily detected using conventional RT-PCR and agarose gel electrophoresis. In an analysis of CK 18 mRNA expression in peripheral venous blood in 13 healthy volunteers, we found that CK 18 mRNA was much less expressed than in cancer cell lines. However, the expression in all samples was at a level which was also detected using conventional RT-PCR. It would thus seem that not only qualitative, but also quantitative analysis, of the target mRNA is important to detect micrometastasis. Quantitative RT-PCR methods will make comparisons of the possible differences in expression levels of the target gene. For clinical applications, much further study is needed.

Topics: Adult; Colorectal Neoplasms; DNA Primers; Electrophoresis, Agar Gel; Esophageal Neoplasms; Genetic Vectors; Glyceraldehyde-3-Phosphate Dehydrogenases; Humans; Keratins; Lung Neoplasms; Male; Reference Values; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Stomach Neoplasms; Transcription, Genetic; Tumor Cells, Cultured

To detect lymph node metastases by immunohistochemistry, where previously undetected by routine histopathology.. Immunostaining was carried out for high- and low molecular weight cytokeratins, and Ber-EP4 in 19 consecutive lung cancer patients who had undergone systematic mediastinal lymph node dissection.. Eleven (58%) epidermoid carcinomas, 6 (32%) adenocarcinomas, and 2 (10%) bronchiolo-alveolar carcinomas were detected. These included 4 (21%) stage IA carcinomas, 6 (32%) stage IB, 6 (32%) stage IIB, 1 (5%) stage IIIB and 2 (10%) stage IV. Immunostaining did not reveal any undetected metastases. Two patients (squamous cell carcinoma T1N0; adenocarcinoma T1N0) had metastases (skeletal; ipsilateral lung) at time of surgery, and one patient (squamous cell carcinoma T2N0) had a regional and systemic relapse 10 months later. Serial sectioning with immunostaining of the lymph nodes from these three patients was also negative.. We conclude that, even with the use of immunostaining, negative lymph nodes will not assure a good prognosis, and different determinants probably exist for lymphatic and hematogenic metastases in non-small cell lung cancer.

Topics: Aged; Aged, 80 and over; Antigens, Surface; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Node Excision; Lymphatic Metastasis; Male; Middle Aged

The aim of this study is to assess the clinical usefulness of serum assays of carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), and CYFRA 21.1 in the diagnosis of squamous cell lung cancer. Sixty patients with squamous cell, and twenty-four patients with nonsquamous cell histology of nonsmall cell lung cancer were enrolled in this study. Serum CEA, SCC, and CYFRA 21.1 levels were obtained by commercially available kits. Upper cutoff levels were 10 ng/ml, 3.5 ng/ml, and 3.5 ng/ml, respectively. In squamous cell lung cancer, percentages and 95% confidence interval (CI) of the patients with elevated levels were as follows: for CEA 23.3% (13-36), for SCC 20.0% (10-32), and for CYFRA 21.1 85.0% (73-93). The positivity rate of CYFRA 21.1 was more significant than CEA and SCC in both squamous and nonsquamous cell lung cancer. None of the markers were significant in differentiating squamous/nonsquamous histology. Only tumor marker CEA was significantly elevated in metastatic squamous cell lung cancer (p=0.004). A novel tumor marker CYFRA 21.1 can be used as a reliable tumor marker in diagnosing squamous cell lung cancer. In addition, CEA has an important role in determining metastatic disease.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Confidence Intervals; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Reproducibility of Results; Serpins; Smoking

A clinicopathological study of 10 patients with pleomorphic carcinoma of the lung.. Histopathological and immunohistochemical staining for keratin, vimentin, Mac387, desmin, actin and S-100 protein were used for this study.. Pleomorphic carcinoma of the lung was found to often occur in males above 50 years of age and with clinical symptoms including cough, expectoration, haemoptysis and chest pain. The most frequent microscopic diagnosis was squamous cell carcinoma, and adenocarcinoma, accompanied by spindle and giant cells. The epithelial component of pleomorphic carcinoma of the lung displayed positivity for keratin and the spindle cells displayed positivity for vimentin. In some cases the neoplastic epithelial component and spindle cells showed positive expression of both keratin and vimentin.. Pleomorphic carcinoma of the lung may display various histopathological changes making it easy to be misdiagnosed as carcinosarcoma. Understanding its pathogenesis and histopathology is important for the diagnosis and differential diagnosis.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Carcinoma; Carcinoma, Giant Cell; Carcinoma, Large Cell; Carcinoma, Squamous Cell; Carcinosarcoma; Diagnosis, Differential; Female; Follow-Up Studies; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Pneumonectomy; Vimentin

Topics: Adult; Antineoplastic Combined Chemotherapy Protocols; Choriocarcinoma; Chorionic Gonadotropin, beta Subunit, Human; Endodermal Sinus Tumor; Fatal Outcome; Humans; Keratins; Lung Neoplasms; Male; Mediastinal Neoplasms; Radiography, Thoracic; Tomography, X-Ray Computed

Considering the current need to improve cost-effectiveness in cancer patient management, a prospective study was undertaken in order to define the optimal combination of bone scan and tumour marker assays in breast and lung cancer strategies, as has been done in the case of prostate cancer. All patients with breast or lung cancer referred to the Nuclear Medicine Department of the Grenoble Teaching Hospital between December 1995 and April 1997 were included. A blood sample was drawn in each case for marker assay (CA15-3 or CEA and CYFRA 21-1) on the same day as the bone scan. Two hundred and seventy-five patients were included: 118 with lung cancer and 157 with breast cancer. With regard to lung cancer, no information useful for guiding bone scan prescription was obtained through CEA and CYFRA 21-1 assays. For breast cancer, the results suggest that in asymptomatic patients, a CA15-3 level of less than 25 U/ml (upper normal value chosen as the threshold) is strongly predictive of a negative bone scan; by contrast, high tumour marker levels are predictive of neoplastic bone involvement. When a doubtful bone scan is obtained in a patient with breast cancer, a normal marker level makes it highly probable that bone scan abnormalities are not related to malignancy.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Bone and Bones; Bone Neoplasms; Breast Neoplasms; Breast Neoplasms, Male; Carcinoembryonic Antigen; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Mucin-1; Predictive Value of Tests; Prospective Studies; Radionuclide Imaging; Radiopharmaceuticals; Technetium Tc 99m Medronate

High serum NSE and advanced tumour stage are well-known negative prognostic determinants of small cell lung cancer (SCLC) when observed at presentation. However, such variables are reversible disease indicators as they can change during the course of therapy. The relationship between risk of death and marker level and disease state during treatment of SCLC chemotherapy is not known. A total of 52 patients with SCLC were followed during cisplatin-based chemotherapy (the median number of tumour status and marker level assessments was 4). The time-homogeneous Markov model was used in order to analyse separately the prognostic significance of change in the state of the serum marker level (NSE, CYFRA 21-1, TPS) or the change in tumour status. In this model, transition rate intensities were analysed according to three different states: alive with low marker level (state 0), alive with high marker level (state 1) and dead (absorbing state). The model analysing NSE levels showed that the mean time to move out of state 'high marker level' was short (123 days). There was a 44% probability of the opposite reversible state 'low marker level' being reached, which demonstrated the reversible property of the state 'high marker level'. The relative risk of death from this state 'high marker level' was about 2.24 times greater in comparison with that of state 0 'low marker level' (Wald's test; P < 0.01). For patients in state 'high marker level' at time of sampling, the probability of death increased dramatically, a transition explaining the rapid decrease in the probability of remaining stationary at this state. However, a non-nil probability to change from state 1 'high marker level' to the opposite transient level, state 0 'low marker level', was observed suggesting that, however infrequently, patients in state 1 'high marker level' might still return to state 0 'low marker level'. Almost similar conclusions can be drawn regarding the three-state model constructed using the tumour response status. For the two cytokeratin markers, the Markov model suggests the lack of a true reversible property of these variables as there was only a very weak probability of a patient returning to state 'low marker level' once having entered state 'high marker level'. In conclusion, The Markov model suggests that the observation of an increase in serum NSE level or a lack of response of the disease at any time during follow-up (according to the homogeneous assumption) was strongly associ

Topics: Antigens, Neoplasm; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoma, Small Cell; Cisplatin; Cyclophosphamide; Epirubicin; Etoposide; Humans; Keratin-19; Keratins; Lung Neoplasms; Markov Chains; Prognosis; Proportional Hazards Models; Prospective Studies; Tissue Polypeptide Antigen

Keratin intermediate filaments are formed in epithelial cells in a cell- and tissue-specific manner, but much remains unknown regarding the mechanisms which control the synthesis of these proteins. We examined the effect of the differentiation modulation agent, bromodeoxyuridine (BrdU), on two human keratin-negative (by immunocytochemistry) lung cell lines, DLKP and H82, and showed immunohistochemically that treatment with 10 microM BrdU over 7 days induced K8 and K18 protein synthesis in both lines. Immunoprecipitation and Western blot analyses revealed low levels of K8 and K18 proteins in untreated cell homogenates. These levels increased following treatment with BrdU for 7 days. K8 and K18 mRNAs were detected by Northern blot and reverse transcriptase polymerase chain reaction analyses in both lines before BrdU treatment, but no increase in mRNA levels was observed in either cell line over 21 days of treatment. This suggests, firstly, that keratin synthesis is normally blocked at a posttranscriptional level in DLKP and H82 cells, and secondly, that BrdU can reverse this block. A549 is a human lung cell line which contains K8 and K18 proteins. Treatment with BrdU increased K8 and K18 protein levels in these cells. No corresponding increase in K8 mRNA levels occurred, while an apparent increase in K18 mRNA levels was detected. HL-60 is a leukaemic cell-line of haematopoietic rather than epithelial lineage which contains K8 and K18 mRNA transcripts prior to BrdU treatment, but does not contain keratin proteins. Again, K8 and K18 mRNA levels remained unchanged during BrdU treatment. However, neither K8 nor K18 proteins were detected following treatment, although BrdU is known to alter expression of other genes in HL-60 cells. BrdU thus appears to act at a posttranscriptional level and in an epithelial-specific manner to reverse a block in keratin synthesis in keratin-negative lung cancer cells and increase synthesis in keratin-positive lung cancer cells. This may represent a regulatory step in early lung development or a mechanism whereby tumour cells downregulate expression of a differentiated phenotype.

Topics: Antimetabolites, Antineoplastic; Bromodeoxyuridine; Carcinoma, Small Cell; Down-Regulation; Humans; Keratins; Lung Neoplasms; Protein Processing, Post-Translational; RNA, Messenger; Tumor Cells, Cultured

No studies about correlation between post-operative half-life of tumor markers and prognosis in lung cancer exist in literature. The aim of our study was to determine the half-life of CEA, TPA, NSE and CYFRA 21-1 in postoperative period after surgery of bronchogenic carcinoma, and to correlate it with the prognosis and survival of the patients.. From March 1997 to March 1998, 35 patients with bronchogenic carcinoma were studied (29 males and 6 females, mean age 64.9 years, range 51-77 and 61.0 years, range 52-77 respectively). The mean follow-up for males was 125.70 days (from 30 to 198) after surgery and for females 125.79 days (from 30 to 180). CEA and NSE were tested by immunoenzymatic automated method, whereas TPA and CYFRA 21-1 were assayed by immunoradiometric techniques. For each patient both the dismission curve and the half-life of considered markers were calculated during follow-up.. A statistically significant difference was found for preoperative values of TPA (p = 0.027) and CYFRA 21-1 (p = 0.025) between SqCLC and adenocarcinoma. The preoperative levels of markers were higher in patients who would develop a relapse, even if statistical significance was not reached. CEA half-life was of 1.4 days, while in patients with a history of relapse or metastatic spreading was 4.5 days. No differences were revealed concerning CYFRA 21-1 between the two groups.. Seriate determination of some markers (CEA and TPA in particular) during postoperative follow-up after surgery for bronchogenic carcinomas can be a useful prognostic tool. Longer follow-up would provide additional informations in order to determine individual predictive threshold between poor and good prognosis.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Female; Half-Life; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Postoperative Period; Prognosis; Time Factors; Tissue Polypeptide Antigen

The present study was designed to clarify the mechanism by which some lung cancer cell lines can produce cytokeratin 19 (CK19) fragment and others cannot. We hypothesized that some lung cancer cell lines which cannot release CK19 express an incomplete sequence of CK19 mRNA. Expression of mRNA was evaluated by RT-PCR using several primer pairs for CK19. CK19 in the culture supernatant was measured by an immuno-radiometric assay. CK19 protein synthesis was evaluated by Western immunoblot and immunohistochemistry. Among 16 lung cancer cell lines, 7 released significant amounts of CK19 in the supernatant. In some cell lines, expression of CK19 mRNA was observed only in some combinations of primers, suggesting that incomplete mRNA was expressed. 3'-RACE analysis detected amplified products of a shorter size compared with normal amplified products in cell lines which expressed incomplete CK19 mRNA, suggesting that 3'-ends of mRNA for CK19 were deleted. Results of Western immunoblot and immuno-histochemical staining using anti-human CK19 monoclonal antibody completely correlated with the results on CK19 levels in culture supernatants as well as with complete expression of mRNA. We conclude that levels of CK19 closely relate to the expression of complete mRNA for CK19.

Topics: Adenocarcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; DNA Primers; Gene Expression Regulation, Neoplastic; Humans; Keratins; Lung Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription, Genetic; Tumor Cells, Cultured

A Sertoli cell carcinoma of the ovary with lung metastases mimicking neuroendocrine carcinoma is presented. Lung metastases frequently occur. Primary and secondary tumors may exhibit similar growth patterns and differentiating primary from secondary tumors may be troublesome. This process may be more difficult when metastases occur from a tumor in which metastases are uncommon and morphologically resemble only a small portion of the primary tumor. We report the case of a 52-year-old woman who underwent resection of a 4,550-g Sertoli cell tumor of the ovary. Histologically, in addition to the characteristic tubular pattern of growth, 5% of the tumor consisted of poorly differentiated areas with tumor cells in sheets, a high mitotic rate, and areas of necrosis. Eleven months after this surgery she presented at a different institution with multiple pulmonary nodules. Microscopic examination of a subsequently resected lung nodule showed histologic findings similar to those of the poorly differentiated areas of the ovarian tumor and initial immunohistochemical studies showed positive staining for cytokeratin, neuron-specific enolase, and focal positivity for synaptophysin. Without knowledge of the ovarian tumor the lung lesion was interpreted as large-cell neuroendocrine carcinoma. On review of the clinical history and comparison with the previous surgical material, however, both tumors showed similar light microscopy and immunohistochemical reactivity, and a final diagnosis of metastatic Sertoli cell tumor was made. Immunohistochemical staining for inhibin revealed weak positivity in the poorly differentiated areas of the ovarian tumor but not in the lung metastasis. This is one of the rare reports of ovarian Sertoli cell tumor metastasizing to the lungs and it emphasizes the importance of complete clinical histories, ancillary studies, appropriate sampling, and review of archival material in such unusual cases.

Topics: Carcinoma, Neuroendocrine; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Inhibins; Keratins; Lung Neoplasms; Middle Aged; Ovarian Neoplasms; Phosphopyruvate Hydratase; Sertoli Cell Tumor; Synaptophysin

Fifty patients with adenocarcinoma of the lung were enrolled in this study, including 20 patients with recurrence and 30 patients without recurrence 1 year after surgery. Serial serum levels of cytokeratin fragment 19 (CYFRA 21-1) and carcinoembryonic antigen (CEA) were measured before the operation and 1 week, 1 month, 3 months, 6 months, 9 months, and 12 months after surgery for the early detection of recurrence. The results revealed that the mean serum values of either CYFRA 21-1 or CEA were significantly higher until 9 and 12 months after surgery in the 20 patients with recurrent adenocarcinoma compared with the 30 patients without recurrent adenocarcinoma. We conclude that CYFRA 21-1 is not a better marker than CEA for early prediction of adenocarcinoma. We conclude that CYFRA 21-1 is not a better marker than CEA for early prediction of adenocarcinoma recurrence in lung.

Topics: Adenocarcinoma; Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Prognosis; Prospective Studies

The aim of this prospective study was to assess the diagnostic value of the imaging modalities (chest radiography, spiral computed tomography (SCT) and high-resolution computed tomography (HRCT)) and the tumour markers (carcinoembryonic antigen (CEA), cytokeratin marker (CYFRA 21-1) and neuron-specific enolase (NSE)) in the differentiation of malignant (MSPLs) from benign solitary pulmonary lesions (BSPLs).. Solitary pulmonary lesions (SPLs) were examined, evaluated and then completely removed by surgery in 104 consecutive patients (MSPLs n = 81, BSPLs n = 23). Chest radiography was performed with frontal and lateral views, SCT was carried out with a slice thickness of 8 mm and HRCT with a slice thickness of 1 mm and a 12-cm field of view. For the tumour marker analysis, serum concentrations were determined 1-3 days prior to surgery by ELISA for CEA and CYFRA 21-1 and by IRMA for NSE using commercially available assay kits. The cut-off values were set at 3 ng/ml (for non-smokers) and 5 ng/ml (for smokers) for CEA, at 3.3 ng/ml for CYFRA 21-1 and at 12.5 ng/ml for NSE.. Using any one of the characteristics with a significance level of P <0.01, the identification of MSPLs using chest radiography showed a sensitivity of 64.2% and a specificity of 82.6%, using SCT a sensitivity of 88.9% and a specificity of 60.9% and using HRCT a sensitivity of 91.4% and a specificity of 56.5%. For the identification of MSPLs using CEA a sensitivity of 27.2% and a specificity of 87.0% (accuracy of 40.4%) was observed. Using CYFRA 21-1 a sensitivity of 19.8% and a specificity of 100.0% (accuracy of 37.5%) and using NSE a sensitivity of 13.6% and a specificity of 100. 0% (accuracy of 32.7%) was found.. Using chest radiography, SCT and HRCT, a precise morphological assessment of the periphery of the pulmonary lesion and the adjacent visceral pleura is necessary to distinguish MSPLs from BSPLs. Tumour markers used alone or in combination with the imaging methods brought no additional benefits, in terms of sensitivity and accuracy, over the diagnostic imaging methods alone. However, the tumour markers exhibited a far superior specificity (100% for CYFRA 21-1 and NSE) compared with the imaging methods.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Lung; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity; Tomography, X-Ray Computed

The poor prognosis of malignant rhabdoid tumor is emphasized and histopathological criteria for distinction from epithelial sarcoma of the vulva are discussed. Immunohistochemical analyses were performed by using nine different antigens including vimentin, cytokeratin, epithelial membrane antigen, carcinoembryonic antigen, desmin, muscle-specific actin, S-100 protein, AP-15, neuron specific enolase. This is the sixth reported case of a malignant rhabdoid tumor of the vulva. The patient died eight months after the initial diagnosis in spite of a combination of surgery, adjuvant chemotherapy and external radiotherapy.

Topics: Adult; Biomarkers, Tumor; Fatal Outcome; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Neoplasm Recurrence, Local; Rhabdoid Tumor; Skin Neoplasms; Vimentin; Vulvar Neoplasms

The morphology, cell growth, antigenic expression and tumorigenicity of cell subpopulations from the A549 lung adenocarcinoma isolated by Percoll gradient separation have been analysed. Four subpopulations were obtained (subpopulations A, B, C and D). Immunocytochemical analysis of several antigens was performed with monoclonal antibodies (MAbs): MUC1 mucin (C595, HMFG1 and HMFG2), MUC5B (PANH2); gp230 (PANH4); carbohydrate antigens including sialyl Lewis x (KM93), Tn antigen (83D4), Lewis y (C14); 5, 6, 8, 17 and 19 cytokeratins and p53. The cell population D tended to form cell aggregates that piled up on the monolayer similar to overgrowth cultures of the A549 parental cell line, whereas A, B and C cell subpopulations formed well spread monolayers. Both parental A549 and subpopulation D secreted abundant mucus. The topographic distribution and secretion production were correlated with tumorigenic assays since only subpopulation D grew in nude mice exhibiting reduced latency period; these characteristics correlated with the fast growth of the subpopulation D in vitro. Immunocytochemical analysis demonstrated that subpopulation D showed greater expression of MUC1 mucin and carbohydrate antigens such as Tn antigen, sialyl Lewis x and Lewis y and less expression of cytokeratins, p53, MUC5B and gp230; conversely, subpopulations A, B and C showed the opposite antigenic profile. Our results illustrate heterogeneity in the A549 cell line; subpopulations A, B and C retained characteristics of more differentiated adenocarcinoma while subpopulation D displayed features of a less differentiated tumor line.

Topics: Adenocarcinoma; Animals; Antibodies, Monoclonal; Cell Separation; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Transplantation; Oligosaccharides; Phenotype; Sialyl Lewis X Antigen; Time Factors; Tumor Cells, Cultured; Tumor Suppressor Protein p53

We examined changes in cytokeratin 19 fragment (CYFRA 21-1) levels in relation to the T factor in 64 non-small cell N2M0 lung cancer patients. Although a correlation between the levels and T factor was found (rho=0.627, p<0.0001), there was no difference between the levels in T3 and T4. Serum CYFRA 21-1 levels increased in the order of the following groups: the limited tumor group (T1+T2: n=28), the group with tumor extending to the pleura or chest wall (T3: n=13), and the group with tumor invading into the mediastinum (T4: n=12). The level was lower in the group with malignant pleural effusion (T4: n=11) than in the group with tumor invading into the mediastinum (6.7+/-4.7 ng/ml vs. 12.2+/-8.1 ng/ml, p=0.0046). We suspect that the presence of malignant pleural effusion is not directly related to the three-dimensional expansion of the tumor and this is a reason why CYFRA 21-1 levels in T4 are not higher than those in T3.

Topics: Antigens, Neoplasm; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Keratins; Lung Neoplasms; Pleural Effusion; Radiometry

It has been reported that cytokeratin 8 (CK8) is expressed in all non-small-cell lung cancers (NSCLC). We hypothesized that antigenic changes of CK8 may occur in some NSCLC cell lines. To prove this, Western immunoblot analysis using anti-human CK8 monoclonal antibodies as well as immunohistological staining of CK8 were performed in NSCLC cell lines. As a result, CK8 which had a higher molecular weight than recombinant CK8 was demonstrated in two of eight NSCLC cell lines. In addition, this CK8 contained antigenic epitopes of CA19-9. This CK8 with higher molecular weight, may have played a role in the process of invasion or metastasis of NSCLC.

Topics: Adenocarcinoma; Animals; Blotting, Western; CA-19-9 Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Carrier Proteins; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Molecular Weight; Neoplasm Proteins; Rabbits; Tumor Cells, Cultured

The aim was to investigate the diagnostic utility of CYFRA 21-1 (cytokeratin 19 fragment) as a tumor marker in pleural effusion and evaluate the value of combining CYFRA 21-1 and carcinoembryonic antigen (CEA) assays as a diagnostic aid in the malignant pleural effusion.. One hundred and twenty-six patients (72 malignant and 54 benign pleural effusion) were included in this retrospective study. The effusion levels of CYFRA 21-1 and CEA were measured using radioimmunometric assay.. The median values of CYFRA 21-1 in benign and malignant pleural effusion are 15 and 70 ng/ml, respectively. Using a cut-off value of 50 ng/ml, defined at 94% specificity, the diagnostic sensitivity of CYFRA 21-1 for non-small cell lung carcinoma (n = 61), squamous cell carcinoma (n = 21), adenocarcinoma (n = 40) and small cell lung cancer (n = 11) was 64, 71, 60 and 18%, respectively. Regardless of cell types, the diagnostic sensitivity of CYFRA 21-1 and CEA in malignant pleural effusion (n = 72) was 57 and 60%, respectively (cut-off value of 10 ng/ml in CEA assay). Combining CEA with CYFRA 21-1, the diagnostic sensitivity may increase up to 72%, which was defined at 89% specificity.. CYFRA 21-1 assay may be a useful tumor marker for discriminating benign from malignant pleural effusion, especially in those of non-small cell lung cancer. The combined use of CEA and CYFRA 21-1 assay in the malignant effusion may increase the diagnostic yield compared with CEA or CYFRA 21-1 alone.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Humans; Keratin-19; Keratins; Lung Neoplasms; Pleural Effusion, Malignant; Radioimmunoassay; Retrospective Studies; Sensitivity and Specificity

Epithelioid trophoblastic tumor (ETT) is a term proposed for an unusual variant of trophoblastic tumor that is closely related to choriocarcinoma but shows monomorphic growth of highly atypical trophoblastic cells instead of the typical dimorphic pattern of choriocarcinoma. We report here 3 cases of ETT, all of which were lung lesions probably originating from uterine trophoblastic disease. The antecedent pregnancies of the 3 cases were hydatidiform mole, invasive mole, and term pregnancy, respectively. The tumors were composed of highly atypical mononucleate cells, which mainly involved alveolar spaces, forming nests with central eosinophilic necrosis. Multinucleate giant cells were found within the nests, but they were fewer in number than in typical choriocarcinoma. The tumors were not associated with extensive hemorrhage or necrosis, except for 1 case, in which the ETT was combined with typical dimorphic choriocarcinoma. Immunohistochemically, multinucleate giant cells and occasional mononucleate tumor cells showed positivity for human chorionic gonadotropin. Staining for human placental lactogen was positive in rare multinucleate giant cells, and in 1 case, tumor cells showed diffuse positivity for placental alkaline phosphatase. Because ETT has a remarkably epithelioid appearance in cytological and architectural features, differentiation from the epithelial malignancies is problematic. Trophoblastic markers are frequently expressed in nontrophoblastic tumors, and reactivity for those markers alone is not sufficient for exclusion of other tumors. Rather, evidence of ETT comes from a combination of morphological features, immunohistochemical study, and clinical history.

Topics: Adult; Chorionic Gonadotropin; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Microscopy, Electron; Middle Aged; Neoplasms, Glandular and Epithelial; Pregnancy; Trophoblastic Neoplasms

Topics: Aged; Antibodies, Neoplasm; Humans; Keratins; Lung Diseases, Interstitial; Lung Neoplasms; Male

The diagnostic value of tumour markers in pleural effusion is not yet clearly defined. CEA (Carcinoembryonic Antigen), CYFRA 21-1 (Cytokeratin 19-Fragment) and TPA-M, a new monoclonal-based radioimmunoassay for TPA (Tissue Polypeptide Antigen), were measured in pleural fluid and sera of 125 consecutive patients who underwent medical thoracoscopy. The group consisted of 79 patients with malignant and 45 with non-malignant pleural effusion and 1 patient without definitive diagnosis, and hence 124 patients were available for assessing the diagnostic value. In pleural fluid based on a specificity of 90% versus benign diseases the sensitivity for CEA was 52.5%; with the maximum achievable specificity of 80% for CYFRA 21-1 the sensitivity was 68% and for TPA-M with 67% the sensitivity was 67%. Based on the cut-off values for these specificities the combined use of the three tumour markers resulted in a sensitivity of 85.7% but with a lower specificity of 59.1%. There is only a limited value for tumour markers in the diagnosis of pleural effusion.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Pleural Effusion; Pleural Effusion, Malignant; Radioimmunoassay; Sensitivity and Specificity; Thoracoscopy; Tissue Polypeptide Antigen

The aim of this prospective study was to assess the diagnostic value of the tumour markers carcinoembryonic antigen (CEA), cytokeratin 19 fragment marker (CYFRA 21-1) and neuron-specific enolase (NSE) in the differentiation of malignant (MSPLs) from benign solitary pulmonary lesions (BSPLs).. Solitary pulmonary lesions (SPLs) were diagnosed using plain radiography and spiral computed tomography (SCT) and then completely removed by surgery in 104 consecutive patients (MSPLs; n = 81, BSPLs; n = 23). The serum concentrations of the tumour markers were determined 1-3 days prior to surgery by ELISA for CEA and CYFRA 21-1 and by IRMA for NSE using commercially available assay kits. The cut-off values were set at 3 ng/ml (for non-smokers) and 5 ng/ml (for smokers) for CEA, at 3.3 ng/ml for CYFRA 21-1 and at 12.5 ng/ml for NSE.. MSPLs were identified with a sensitivity between 13.6 and 45.7%, a specificity between 87.0 and 100% and an accuracy between 32.7 and 54.8%. Using the tumour markers alone, the highest sensitivity (27.2%) and accuracy (40.4%) was found with CEA, the highest specificity (100%) with CYFRA 21-1 and with NSE. Primary lung cancers (n = 39) were identified with a sensitivity between 17.9 and 61.5%, a specificity between 87.0 and 100% and an accuracy between 48.4 and 71.0%. Using the tumour markers alone, the highest sensitivity (35.9%) and accuracy (59.7%) was found with CYFRA 21-1, the highest specificity (100%) with CYFRA 21-1 and with NSE. The combination of all three tumour markers resulted in a greater sensitivity and greater diagnostic accuracy but a loss in specificity compared with CYFRA 21-1 and NSE.. The use of the tumour markers alone or in combination showed a low sensitivity and low accuracy for the diagnostic differentiation of MSPLs from BSPLs and primary lung cancers from BSPLs. However, both CYFRA 21-1 and NSE exhibited a specificity of 100% and may be useful complements to standard clinical imaging methods.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Diagnosis, Differential; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Prospective Studies; Sensitivity and Specificity; Solitary Pulmonary Nodule

The detection of metastasizing single tumor cells has so far been difficult. Using a monoclonal antibody (MAb A45-B/B3) recognizing human cytokeratin, we identified immunocytochemically single tumor cells and micrometastases in patients (n = 24) with nonsmall cell lung cancer at the time of surgery of the primary tumor. The cytokeratin-positive cells (1-14/5 x 10(5) cells) in the bone marrow samples of 9 (9/24) patients were found. We also found a garland-like cluster, which consists of seven cancer cells and two closely connected tumor cells from one bone marrow sample. These results indicate that this technique can be used as a early diagnostic technique of bone marrow micrometastasis in the patient with the nonsmall cell lung cancer.

Topics: Animals; Antibodies, Monoclonal; Bone Marrow Neoplasms; Carcinoma, Non-Small-Cell Lung; Epitopes; Humans; Hybridomas; Immunohistochemistry; Keratins; Lung Neoplasms; Mice

It recently became evident that isolated tumor cells undetectable by conventional tumor staging are frequently present in bone marrow of patients with apparently localized non-small cell lung cancer (NSCLC). The clinical relevance of this minimal hematogenous tumor cell dissemination is under vigorous debate.. For tumor cell detection in the bone marrow, we used monoclonal antibody CK2 against the epithelial intermediate filament protein cytokeratin 18. The influence of a positive bone marrow finding on clinical outcome was studied in 139 patients with NSCLC postoperatively staged as pT1-4, pN0-2, M0, and R0 after a median follow-up of 66 months (range 48 to 74 months).. Cytokeratin-18-positive cells in bone marrow were demonstrated in 83 (59.7%) patients at the time of primary surgery and in 6 of 12 representative patients analyzed twice 3 to 18 months after surgery. In patients without histopathological lymph node metastases (pN0; n = 66), the occurrence of 2 or more tumor cells in bone marrow at primary surgery was a strong and independent predictor for overall survival (p = 0.007) in univariate analysis. The multivariate analysis showed a 2.8 times increased risk for shorter survival in patients with disseminated tumor cells versus patients without such cells. Four of the 6 patients with a positive cytokeratin status after surgery developed a tumor recurrence 11 to 44 months after the operation, while none of the patients with a negative bone marrow at all time intervals showed a tumor relapse.. Minimal residual bone marrow involvement is an independent prognostic factor for overall survival in patients with node-negative NSCLC, which may help to identify patients in need of an adjuvant systemic therapy. The postoperative persistence or reappearance of tumor cells in bone marrow indicates that these are not only shed cells but rather represent true micrometastasis.

Topics: Bone Marrow; Bone Marrow Neoplasms; Carcinoma, Non-Small-Cell Lung; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Middle Aged; Multivariate Analysis; Risk Factors; Survival Rate

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Keratins; Lung Neoplasms; Practice Guidelines as Topic; Prognosis; Pulmonary Medicine; Societies, Medical

Small cell lung carcinoma (SCLC) is the most aggressive of lung tumors, metastasize widely and are virtually incurable by surgical means. Therefore, the classification of lung cancer into SCLC and non-small cell lung carcinoma is essential for disease prognosis and treatment. For this purpose we have compared the immunohistochemical distribution of different cytoskeletal proteins as tumor markers. Analysis was performed by using of monoclonal antibodies directed against cytokeratins, neurofilaments, betaIII-tubulin, epithelial membrane antigen and neuron-specific enolase. Our results indicate that keratin and epithelial membrane antigen are reliable epithelial markers for SCLC. In addition, the positive staining with monoclonal antibodies TU-20 against betaIII-tubulin and neuron-specific enolase was found in some cases of SCLC. We suggest, that these antibodies could be a useful tool for complex immunohistochemical diagnosis of SCLC.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Small Cell; Cytoskeletal Proteins; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mucin-1; Phosphopyruvate Hydratase; Tubulin

An anticorpal panel formed by Calretinin, Ber-EP4, Keratin and CD 68 has been applied to differentiate mesothelial hyperplasia/mesothelioma from metastastic carcinoma in cavity effusions. The study was performed in 86 cases in which paraffinated cell-blocks were obtained. All cases positive for malignancy had histological confirmation. The series included 2 malignant mesothelioma, 54 reactive mesothelial hyperplasia, and 30 metastatic carcinoma. All cases of reactive mesothelial showed strong cytoplasmatic positivity for Calretinin, and hyperplasia negativity for Ber-EP4. The 2 cases of mesothelioma were positive for Calretinin and negative for Ber-EP4. In contrast, all cases of metastatic carcinoma had membrane or cytoplasmatic positivity for Ber-EP4. The sensitivity (100%) and specificity (100%) of reactivity for Calretinin and Ber-EP4 showed that the immunocytochemical results on paraffin embedded material from cavity effusions are very reliable, so that the tested immunocytochemical panel can be useful in cytological differential diagnosis between reactive mesothelial hyperplasia/mesothelioma and metastatic carcinoma.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Antigens, Surface; Ascites; Biomarkers, Tumor; Calbindin 2; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Macrophages; Nerve Tissue Proteins; Pleural Effusion, Malignant; S100 Calcium Binding Protein G; Sensitivity and Specificity

Recently, cytokeratins (CK) were studied as tumor markers for many carcinomas. In lung cancer they appeared to be useful in distinguishing primary from secondary tumors, in histological typing as well as in evaluating patient's prognosis. However, the results have yet to be conclusive. In this study, expression of CK7, CK10/13, CK18, CK19, CK20 was investigated in a group of 72 surgically resected specimens of lung including 31 adenocarcinomas, 30 squamous cell carcinomas and 11 neuroendocrine carcinomas. Cytokeratin immunophenotypes were analyzed in comparison to histological characteristics of tumors, TNM stages and patients survival.. CK7, CK10/13 and CK18 can be used in distinguishing the lung adenocarcinomas from the lung squamous cell carcinomas: CK7(+), CK10/13(-), CK18(+) for adenocarcinomas; CK7(-), CK10/13(+), CK18(-) for squamous cell carcinomas. Relatively higher CK7 and CK18 immunostaining rates of the squamous cell carcinomas with high keratinization, with high percentage of dead cells and with late stages of disease suggested their prognostic significance but it was not confirmed when comparing different survival groups. Both adenocarcinomas and squamous cell carcinomas were stained strongly with antibody against CK19 (90.3% and 86.7% respectively) but much less with anti-CK20 antibody (9.7% and 3.3% respectively). In general, neuroendocrine tumors of the lung were non-reactive for these cytokeratins except CK18, among them all carcinoid tumors expressed CK18 abundantly.

Topics: Adenocarcinoma; Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Molecular Weight; Neuroendocrine Tumors

To evaluate the clinical usefulness of CYFRA21-1 as a serum tumor marker in diagnosis, evaluation of clinical status and prognosis of patients with non-small-cell lung cancer (NSCLC).. CYFRA21-1 in 126 serum samples of NSCLC patients was measured by radioimmunoassay. Of the 126 samples, 71 were collected before treatment, 29 at two weeks after operation, and 26 after 15-18 months follow-up.. CYFRA21-1 was positive in 49.3% of the NSCLC patients who had not received any treatment. The positive rate was 69.2% for squamous-cell carcinoma, and 25.0% for adenocarcinoma. The serum level of CYFRA21-1 was significantly higher in squamous-cell carcinoma than in adenocarcinoma (P < 0.0001). The serum level of CYFRA21-1 well correlated and increased with TNM staging (P = 0.0001). The level decreased significantly within two weeks after surgical operation in 29 cases (P = 0.0005). On follow up for 15-18 months, no change in CYFRA21-1 level was observed in 16 patients whose disease was stable, while there was significant increase in 10 patients with progressive disease.. CYFRA21-1 is a soluble fragment of cytokeratin 19 in serum of patients with NSCLC. It can be used as a useful tumor marker of NSCLC.

Topics: Adolescent; Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Child; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging

We examined the incidence and association of bronchiolization of the alveoli with non-small cell lung cancer in lung resection specimens from 2 patient groups: those with non-small cell lung cancer and those diagnosed with a variety of non-neoplastic lung conditions. We observed marked variation in bronchiolization of the alveoli morphology ranging from normal to severely atypical and developed a classification scheme based on growth pattern, cell number and cytologic criteria. Patterns of differentiation, proliferation and growth factor receptor and oncogene expression were studied using immuno-histochemical and in situ hybridization techniques. While low-grade (0-I) bronchiolization of the alveoli lesions demonstrated markers similar to normal bronchiolar epithelium, a significant decrease in the Clara cell 10 kDa protein and tubulin and an increase in surfactant protein-A expression were observed in high-grade (II-III) lesions. Focal p53 expression was detected in 2 high-grade lesions, while c-myc mRNA and cJun protein were observed in all grades. No correlation was observed between bronchiolization of the alveoli incidence and histologic tumor type. A comparison of marker expression in lesions and tumors from the same case revealed a negative correlation between cytokeratin-14 and c-erbB-2 immuno-reactivity. Only one bronchialization of the alveoli lesion was found in the non-neoplastic patient group. We conclude that up to 12% of non-small cell lung cancer resection specimens contain bronchiolization of the alveoli lesions which exhibit altered morphology and patterns of differentiation.

Topics: Bronchi; Carcinoma, Non-Small-Cell Lung; Cell Differentiation; Humans; Immunoenzyme Techniques; Keratins; Lung Diseases; Lung Neoplasms; Proliferating Cell Nuclear Antigen; Proteins; Proteolipids; Proto-Oncogene Proteins; Pulmonary Alveoli; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants; Receptors, Growth Factor; Uteroglobin

In this report, the cytological features and differential diagnosis of the metastasis from and subsequent local recurrence of an unusual case of malignant (metastatic) ameloblastoma are described, with histological confirmation. Characteristic cytological findings included fibrovascular central cores surrounded by palisading crowded basaloid or columnar cells or both and rosette-like structures of tumor cells with central fibrillary material. Keratin debris in the background and cystic cavities were prominent components of the metastatic ameloblastoma. The basaloid cells showed scant-to-absent cytoplasm, round-to-oval to tear-shaped nuclei, rare longitudinal nuclear grooves, single or multiple nucleoli, and smooth-to-clefted nuclear contours. No features to predict malignant behavior were identified (abundant mitotic activity, necrosis, nuclear pleomorphism). The cytological features of ameloblastoma appear to be characteristic enough to allow definitive diagnosis. However, since the cytology of this tumor is underreported in the literature, the unwary observer could easily misdiagnose it, especially at metastatic sites.

Topics: Ameloblastoma; Biopsy, Needle; Diagnosis, Differential; Female; Humans; Keratins; Lung Neoplasms; Mandibular Neoplasms; Middle Aged; Neoplasm Recurrence, Local

Disseminated epithelial tumor cells have been detected in the bone marrow and blood of cancer patients by means of immunocytochemical or immunofluorescent staining of cytocentrifuge slides, multiparameter flow cytometry, and reverse transcriptase-polymerase chain reaction. However, it is hardly possible using such methods to detect tumor cells at a frequency below 10(-6). To increase the sensitivity of these detection techniques we have developed a new technology for the enrichment of disseminated epithelial tumor cells from hematopoietic cell samples by high-gradient magnetic cell sorting (MACS). Cells are permeabilized and fixed and carcinoma cells are magnetically labeled specifically with an anti-cytokeratin 8 monoclonal antibody (mAb) directly conjugated to superparamagnetic microbeads. Magnetically labeled cells are enriched on high-gradient magnetic columns. Tumor cells are detected in the enriched cell fraction by flow cytometry, fluorescence microscopy, or immunocytochemisty. In this study we demonstrated the method using a model system in which five to 5,000 cells from a breast cancer cell line were seeded into blood cell samples from a healthy donor containing 1.2 x 10(8) leukocytes. Tumor cells were 10,477+/-4242 (n=25)-fold magnetically enriched, and 57.7%+/-16.9% (n=33) of the initially seeded tumor cells were recovered. Applying the method to 20-40 mL blood samples from patients with advanced carcinomas of the breast, prostate, colon, rectum, or lung, we were able to detect between one and 6.8 x 10(4) cytokeratin-expressing tumor cells in 21 of 34 patients. This corresponds to frequencies of tumor cells between 6.8 x 10(-9) and 1.1 x 10(-3) among nucleated cells in the original sample. Enriched tumor cells were further analyzed for expression of tissue-specific and prognostic markers such as breast mucin glycoproteins, erbB2, and CD44v6 for additional characterization and to confirm their tumor origin. The technique described could become a valuable tool for the quantification and molecular characterization of metastatic carcinoma cells in hematopoietic tissue, and may ultimately prove useful in the diagnosis, prognosis, and monitoring of patients with carcinoma.

Topics: Adult; Aged; Breast Neoplasms; Carcinoma; Cell Separation; Colorectal Neoplasms; Epithelial Cells; Female; Humans; Hyaluronan Receptors; Immunohistochemistry; Immunologic Techniques; Keratins; Lung Neoplasms; Magnetics; Male; Middle Aged; Mucins; Neoplasm Metastasis; Prostatic Neoplasms; Receptor, ErbB-2; Tumor Cells, Cultured

The purpose of this study is to evaluate keratin expression in neuroendocrine cells and carcinoid tumor of the gastrointestinal tract and lung. Neuroendocrine cells in the lung and the gastrointestinal tract reacted with low-molecular keratin strongly, but not with high-molecular keratin. The low-molecular keratin expression of carcinoid tumor in the lung and the gastrointestinal tract, which varied in degree, were identical to that of neuroendocrine cells. Keratin 8 and 18 were found abundantly in neuroendocrine tumor and carcinoid tumor of the gastrointestinal tract, and in a lesser amount in carcinoid tumor of the lung, whereas keratin 19 was found in all cases with variable amounts. Based on these results, the combination of antibodies for several types of keratin is useful for examining keratin expression as a marker of epithelium in carcinoid tumor.

Topics: Adult; Biomarkers, Tumor; Carcinoid Tumor; Female; Gastrointestinal Neoplasms; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged

An autopsied case of an 80-year-old man with spindle cell carcinoma of the gingiva is reported. The tumor was polypoid and mostly composed of a sarcomatous proliferation of spindle cells with a small focus of squamous cell carcinoma at the stalk portion. The carcinoma metastasized to a cervical lymph node, lungs and pleura with extension to the diaphragm. In the metastatic lymph node, the squamous cell component was more prominent than the spindle cell one, while only anaplastic pleomorphic carcinoma cells were found in the lungs. The spindle or anaplastic cells were immunohistochemically positive for vimentin and carcinoembryonic antigen (CEA) but not for other epithelial antigens. We have concluded that the sarcomatoid component arose from the oral squamous cell carcinoma by a metaplastic process. This is the first case report of an oral spindle cell carcinoma examined by autopsy.

Topics: Aged; Aged, 80 and over; Anaplasia; Autopsy; Carcinoembryonic Antigen; Carcinoma; Carcinoma, Squamous Cell; Diaphragm; Gingival Neoplasms; Humans; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Metaplasia; Mucin-1; Neoplasms, Multiple Primary; Pleural Neoplasms; Vimentin

Villin (V) is a glycoprotein of microvilli associated with rootlet formation. Most colonic adenocarcinomas have a V positive (+), cytokeratin (CK) 20 (+), CK7-negative (-) immunophenotype; most lung adenocarcinomas have a CK20(-), CK7(+) immunophenotype. The reports of villin immunoreactivity in lung adenocarcinoma range from 6% to 68% in studies using various fixations and varied anti-villin antibodies. Some lung adenocarcinomas have microvilli with rootlets leading to possible diagnostic confusion with metastatic colonic adenocarcinoma to lung. Nine primary lung adenocarcinomas with rootlets on ultrastructure (including four bronchioloalveolar carcinomas [BAC]), four metastatic lung adenocarcinomas with rootlets, nine metastatic colon adenocarcinomas to lung, and 10 randomly selected lung adenocarcinomas without rootlets (including five BAC), were immunostained with monoclonal antibodies to villin (1D2C3), CK7 (OV-TL12/30), and CK20 (Ks20.8) using a streptavidin peroxidase technique with heat-induced epitope retrieval. All primary lung adenocarcinomas with rootlets were CK7(+) CK20(-), and six of nine (67%) were V(+). Cytoplasmic villin positivity occurred in a diffuse--five of nine (56%), focal--two of nine (22%), or brush border pattern--two of nine (22%). Two of four metastatic lung adenocarcinomas with rootlets were V(+). One metastatic lung adenocarcinoma had a CK7(+), CK20(+), V(-) phenotype. All metastatic colonic adenocarcinomas were V(+), CK20(+), CK7(-), and 1 of 10 (10%) lung adenocarcinomas without rootlets was V(+), and all 10 were CK20(-), and CK7(+). In summary, villin positivity is more common in lung adenocarcinoma with rootlets (67%) than those without rootlets (10%). AU primary lung adenocarcinomas were CK7(+), CK20(-). The combination of villin, CK 7, and CK 20 is helpful in differentiating metastatic colon adenocarcinoma from lung adenocarcinoma with rootlets.

Topics: Adenocarcinoma; Biomarkers, Tumor; Calcium-Binding Proteins; Carrier Proteins; Colonic Neoplasms; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lung Neoplasms; Microfilament Proteins; Microscopy, Electron; Microvilli

Lipocortin 1 (annexin I) is a calcium- and phospholipid-binding annexin protein which can be externalised from cells despite the lack of a signal sequence. To determine its cellular distribution lipocortin 1 in A549 human lung adenocarcinoma cells was localised by light- and electron-microscopic immunocytochemistry and by cell fractionation and western blotting. Lipocortin 1 immunoreactivity is concentrated in prominent patches associated with the plasma membrane. The intensity of these patches varied with the confluence and duration of the culture and was not detectably diminished by an EDTA wash before fixation. Tubulin and cytokeratin 8 were colocalized with lipocortin 1 in the patches. Within the cells lipocortin 1 was distributed throughout the cytoplasm. Electron microscopy revealed prominent immunoreactivity along the plasma membrane with occasional large clusters of gold particles in contact with the membrane surface of the cells; within the cytoplasm the membrane of some vesicle/vacuole structures and some small electron-dense bodies was immunoreactive, but no immunogold particles were associated with the multilamellar bodies. Subcellular fractionation, extraction and western blotting showed that lipocortin 1 in the membrane pellet was present as two distinct fractions; one, intimately associated with the lipid bilayer, which behaved like an integral membrane protein and one loosely attached which behaved like a peripheral membrane protein. The results show that a substantial amounts of lipocortin 1 is concentrated in focal structures associated with and immediately beneath the plasma membrane. These might form part of the mechanism by which lipocortin 1 is released from the cells.

Topics: Adenocarcinoma; Annexin A1; Biological Transport; Blotting, Western; Cell Compartmentation; Cell Membrane; Cytochalasin D; Cytoplasm; Cytoskeleton; Fluorescent Antibody Technique; Humans; Keratins; Lung Neoplasms; Membrane Proteins; Microscopy, Immunoelectron; Microtubules; Nucleic Acid Synthesis Inhibitors; Tubulin; Tumor Cells, Cultured

Serologic identification of the fragment of cytokeratin 19 known as CYFRA 21-1 has been used for early detection of squamous cell carcinoma of the lung. The sensitivity of the CYFRA 21-1 assay in detecting oral cancers is lower than that in detecting lung cancers. To clarify the reason for this, we compared the cytokeratin expression in these cancers, with special reference to cytokeratin 19.. Oral squamous cell carcinomas and lung squamous cell carcinomas were immunostained with cytokeratin 19, cytokeratin 10, and cytokeratin 13 antibodies. Staining intensity was scored on a graduated scale from 0 to 4.. With respect to cytokeratin 19, the stainings of all lung cancers were scored as 4, which indicates a greater expression of cytokeratin 19 than is seen in oral cancers (p < 0.01). With an average cytokeratin 19 staining score of 1.67, oral cancers ranked lowest among the antibodies. Squamous cell carcinomas of the maxillary sinus arising from pseudostratified ciliated epithelium were highly expressive of cytokeratin 19. A marker for keratinizing cells (cytokeratin 10) and a marker for squamous cells (cytokeratin 13) were expressed more frequently and intensely in oral cancers (p < 0.01) than in lung cancers (p = 0.019).. From the viewpoint of immunohistochemistry, cytokeratin 19 was found to be a tumor marker with low specificity and sensitivity in oral cancers. The staining results suggested that poor expression of cytokeratin 19 by oral squamous cell carcinoma may result in a low serum value of CYFRA 21-1 in patients with this condition.

Topics: Antibodies; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Coloring Agents; Epithelial Cells; Epithelium; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratin-19; Keratins; Lung Neoplasms; Maxillary Sinus Neoplasms; Mouth Neoplasms; Sensitivity and Specificity

Peripheral pulmonary adenocarcinomas (PPAs) often demonstrate a bronchioloalveolar component, with or without glandular differentiation. PPAs can be nondescript, mucinous, or show features of Type II pneumocytes. Particularly, mucinous lung carcinomas can resemble gastrointestinal metastases. Previous reports suggested that patterns of keratins 7 (K7) and 20 (K20) differ in pulmonary tumors versus enteric metastases. These studies, however, often failed to specify the precise morphotypes of PPA. Thus, we undertook this evaluation of PPAs with different histologic images. Thirty-nine cases were retrieved from institutional files; all were confirmed as primary tumors by clinicopathologic and radiographic review. Cases were classified as Type I (mucinous) bronchioloalveolar carcinoma (BAC1); Type II (nonmucinous) bronchioloalveolar carcinoma (BAC2); conventional PPA with BAC1-like areas (PPA1); or conventional PPA with BAC2-like foci (PPA2). Immunostains were performed for K7, K20, carcinoembryonic antigen, CA19-9, tumor-associated glycoprotein-72, surfactant apoprotein-A, and the c-erbB-2 peptide. BAC1 and PPA1 failed to express surfactant apoprotein-A, and BAC2 also consistently lacked K20, whereas 28% of PPA2 lesions were labeled for K20. All of the other determinants, however, were seen in variable proportions in each subgroup of PPA. Primary BAC1 and PPA1 resembled enteric adenocarcinomas immunophenotypically; on the other hand, BAC2 demonstrated a pattern of protein expression similar to that of Type II pneumocytes. PPA2s are a diverse group of neoplasms, and a subset of PPA2 does show K20 reactivity, as would be expected in metastatic enteric carcinomas. Thus, immunohistochemical data on PPAs must be interpreted carefully and only in clinicopathologic context. With respect specifically to primary pulmonary mucinous tumors, there still seems to be no uniformly reliably marker that will always allow the exclusion of metastatic enteric tumors.

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Diagnosis, Differential; Glycoproteins; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms

The utility of cytokeratin fragment (Cyfra) 21-1, a new tumor marker, was investigated in 100 patients with lung cancer. Sandwich enzyme immunoassay detected Cyfra 21-1 in the sera of 60% of patients. Sensitivity of this marker was especially high (86.4%) for squamous cell carcinoma, exceeding that of a similar marker, squamous cell carcinoma antigen (SCC, 54.5%). In contrast, sensitivity of Cyfra 21-1 was relatively low for adenocarcinoma (52.6%) and for small cell carcinoma (50%). We conclude that Cyfra 21-1 is of value in diagnosis of lung cancer, particularly squamous cell carcinoma.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Sensitivity and Specificity

Detection and quantitation of circulating cancer cells in peripheral blood may improve cancer staging and monitoring. This study explored the feasibility of using circulating cancer cell detection in peripheral blood for the rapid assessment of chemotherapeutic response. Cytokeratin 19 mRNA was amplified by nested reverse transcriptase-PCR in the peripheral blood of 29 healthy volunteers, 33 pneumonia patients, and 86 lung cancer patients. Circulating cancer cells in the peripheral blood were semiquantitatively determined by taking the ratio of cytokeratin 19 band intensity from the second round of nested PCR to the glyceraldehyde-3-phosphate dehydrogenase band intensity from the first round of PCR amplification. The detection limit of the method was 1 cancer cell in 107 peripheral blood mononuclear cells. The positive detection rate was 40% for lung adenocarcinoma patients of all stages, 41% for squamous carcinoma patients of all stages, and 27% for small cell lung cancer patients. Only one control sample from a pneumonia patient showed a positive result (1.6%). The quantitative method reliably and sensitively estimated cancer cell numbers in the peripheral blood of lung cancer patients. Serial measurement of the relative number of circulating cancer cells correlated with the tumor burden and treatment response of patients. This method may help rapidly assess the efficacy of anticancer treatment, redefine cancer staging, and facilitate the design of better therapeutic strategies for the treatment of cancer patients.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Feasibility Studies; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplastic Cells, Circulating; Polymerase Chain Reaction; Sensitivity and Specificity

The aim of this study was (i) to determine predictive factors of a complete response to chemotherapy in small cell lung cancer (SCLC) and predictive factors of an objective response in non-small cell lung cancer (NSCLC) and (ii) to determine whether prognostic factors are different with regard to treatment response and survival. Ninety-nine patients with SCLC and two hundred and two patients with NSCLC received chemotherapy. The following variables were recorded prior to treatment: tumor, node, metastasis status, performance status, body weight loss, blood leukocyte count, serum sodium, serum albumin, lactate dehydrogenase (LDH), alkaline phosphatase, serum NSE, serum TPS, and serum CYFRA 21-1. Tumor response was analyzed at the 10th week. Analysis of survival were done using the landmark method. Hazard ratios of the significant prognostic variables of survival were calculated using the Cox's model. Odds ratios of the significant predicting factors of response were calculated by stepwise logistic regression. In SCLC, the significant determinants of poor survival were: lack of complete response (HR: 2.04), weight loss (HR: 1.76), high serum LDH level (HR: 1.64), and high serum TPS level (HR: 2.47). A high serum TPS level was the only factor among those studied able to predict lack of achievement of complete response (OR: 0.39). In NSCLC, significant determinants of poor survival were: no objective response (HR: 2.28), poor performance status (HR: 2.52), presence of metastases (HR: 1.51), and high serum CYFRA 21-1 level (HR: 1.84). On the other hand, a high serum TPS level (OR: 0.50), the presence of metastases (OR: 0.45), and a leukocyte blood count over 10,000/microl (OR: 0.43) were independent determinants for a patient not to achieve an objective response. We concluded that the predictive factors of complete response in SCLC remain to be defined. On the other hand, in NSCLC three variables contribute to the prediction of an objective response. Finally, determinants of survival differ from predictive factors of response.

Topics: Analysis of Variance; Antibiotics, Antineoplastic; Antigens, Neoplasm; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Cisplatin; Cyclophosphamide; Etoposide; Humans; Keratin-19; Keratins; L-Lactate Dehydrogenase; Lung Neoplasms; Phosphopyruvate Hydratase; Prognosis; Regression Analysis; Tissue Polypeptide Antigen

An extremely rare case of epithelial-myoepithelial carcinoma (EMC) of a lobar bronchus in a 47-year-old female is reported. Grossly, the tumor formed a polypoid mass obstructing the bronchial lumen. Microscopically, it was composed of two cellular types--epithelial cells with eosinophilic cytoplasm and clear myoepithelial cells. Numerous tubules formed by an inner epithelial and outer myoepithelial layer were found. Focally, the tumor showed solid growth of clear cells. Prominent hyalinization of the stroma was found. The nature of the cells was confirmed by positive expression of cytokeratins and epithelial membrane antigen in epithelial cells and vimentin and smooth muscle actin in myoepithelial cells. Differential diagnosis of EMC includes a broad spectrum of salivary gland-type tumors. Furthermore, metastases of clear cell carcinoma of the kidney or thyroid, clear cell ("sugar") tumor of the lung, glandular form of carcinoid, bronchioalveolar adenocarcinoma with myoepithelial cells and pulmonary adenosquamous carcinoma with amyloid-like stroma must be distinguished from EMC. The tumor has neither recurred nor metastasised, a fact supporting the current opinion, that EMC is a tumor of low grade malignancy.

Topics: Actins; Adenocarcinoma; Bronchial Neoplasms; Carcinoma; Diagnosis, Differential; Female; Humans; Immunoenzyme Techniques; Keratins; Kidney Neoplasms; Lung Neoplasms; Middle Aged; Salivary Gland Neoplasms; Thyroid Neoplasms

Merkel cell carcinoma (MCC) has a small cell variant, indistinguishable in hematoxylin-eosin sections from metastatic small cell carcinoma of the lung (SCCL). To investigate whether intermediate filament expression is helpful in this distinction, 17 MCCs of the small cell type were examined for cytokeratin, as well as neurofilament protein immunostaining, and compared with 59 intermediate-type MCCs and 22 SCCL. With a pan-cytokeratin cocktail (cytokeratin 1-8, 10, 13-16, 19), most (39 of 55) intermediate-type tumors and, more important, 11 of 16 cases of the small cell variant exhibited focal paranuclear staining with dot-like positivity, crescentic positivity, or both. A combined focal (dot-like/crescentic) and diffuse cytoplasmic pan-cytokeratin staining was seen in additional 8 of 55 intermediate and 4 of 16 small cell MCCs. Cytokeratin 20 also evoked focal cytoplasmic staining and occasionally focal and diffuse positivity in the MCCs, irrespective of the subtype. Exclusively diffuse cytokeratin 20 patterns did not occur. Conversely, most SCCL showed a diffuse expression of pancytokeratin, and all cases remained cytokeratin 20 negative. When neurofilament protein was applied, approximately half of the MCCs (25 of 40), including 7 of 11 of the small cell variant, were positive, whereas all SCCL were negative. In conclusion, the cytokeratin and neurofilament protein patterns of small cell MCCs are identical to the pattern of intermediate MCCs but differ from the profile of SCCL, which may help in the differential diagnosis.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Merkel Cell; Carcinoma, Small Cell; Diagnosis, Differential; Female; Genetic Variation; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratins; Lung Neoplasms; Male; Middle Aged; Neurofilament Proteins; Phenotype; Skin; Skin Neoplasms

Simultaneous detection of two antigens by immunostaining usually requires primary antibodies from two different species or a hapten modification of one of the antibodies if they are from the same species. A novel double staining method is described for immunodetection of two independent antigens using two mouse monoclonal antibodies. The principle of the method is the following: The first antigen is detected by a monoclonal antibody that is diluted so extensively that it cannot be recognized with conventional detection systems. A highly sensitive biotin-tyramide amplification system is used to identify this antibody. The second antigen is stained with a monoclonal antibody by dilution and detected by conventional immunostaining. The method was tested for both alkaline-phosphatase staining on paraffin sections and immunofluorescence staining on cultured cells in cytospin preparation. The absence of cross-reaction in the former system was demonstrated by the mutually exclusive detection of T- and B-cells in human lymph nodes or T-cells and carcinoma cells in nasopharyngeal carcinoma biopsies. Similarly, the EBV encoded EBNA2 and ZEBRA proteins showed a mutually exclusive staining by immunofluorescence on B95-8 cells. The method could be used to demonstrate co-expression of two independent antigens in the same cells, such as PCNA and keratin in carcinoma cells in paraffin sections and for EBNA2 and LMP1 EBV proteins in immunofluorescence preparations of B95-8 cells.

Topics: Alkaline Phosphatase; Animals; Antibodies, Monoclonal; Antigens; Antigens, CD; Antigens, Viral; Biotin; Cross Reactions; Fluorescent Antibody Technique; Herpesvirus 4, Human; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Lymphoid Tissue; Mice; Nasopharyngeal Neoplasms; Paraffin Embedding; Proliferating Cell Nuclear Antigen; Tumor Cells, Cultured; Tyramine

The aim of this study was to evaluate the diagnostic value of a new tumour marker, cytokeratin fragment 19 (CYFRA 21-1), in bronchoalveolar lavage fluid (BALF) for the diagnosis of lung cancer. The cross-sectional study included 36 patients with lung cancer, 19 with benign lung diseases and 13 control subjects. In the group with cancer, BAL was performed in the cancer-involved lung and in the opposite lung. Results in BALF were expressed both as absolute concentrations (ng ml-1) and referred to total protein (TP) (ng mg-1 TP), and results in plasma were expressed in ng ml-1. In BALF, there was no significant different between cancer and control groups. Using the 95th percentile of levels obtained in benign lung disease in BALF (specificity 95%) as the cut-off point, the sensitivity of CYFRA 21-1 was 13%. Positive and negative predictive values (PPV and NPV) at different pretest probabilities, and positive and negative gains were obtained applying a Bayesian analysis. Results showed low positive gains for PPV (maximal increase of 22%) and almost none for NPV (negative gains < 5%). In plasma, CYFRA 21-1 provided a sensitivity of 65%. The combination of BALF and plasma tumour marker levels showed a sensitivity of 69%. Therefore, measurement of CYFRA 21-1 in BALF has poor diagnostic value in lung cancer.

Topics: Antigens, Neoplasm; Bayes Theorem; Biomarkers, Tumor; Bronchoalveolar Lavage Fluid; Cross-Sectional Studies; Female; Humans; Keratin-19; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity

Merkel cell carcinomas (MCC) not infrequently involve the periorbital region and the eyelids. Clinically, they are relatively characteristic but often unsuspected. Histologically, MCC are often misdiagnosed as lymphoma, melanoma, or metastatic small cell carcinoma of the lung (SCCL).. We present clinical, histological, and immunohistochemical data on six eyelid cases (all females; age 63-102 years; one with concomitant CLL) from our files of 77 MCC with special respect to differential diagnosis. For comparison, 22 SCCL were analyzed. Immunohistochemistry was done with antibodies against pan-cytokeratin (pan-CK), cytokeratin-20 (CK-20), neurofilament protein (NF), neuron-specific enolase (NSE), chromogranin (CHR), and S100 protein (S100).. Morphologically, five of six MCC were prototypic, one was of the small cell variant. Immunohistochemically, dot-like positivities for pan-CK and CK-20 were seen in all six MCC, and for NF in five tumors. None of the 22 SCCL stained positively for CK-20 or NF but 21/22 cases were positive for pan-CK. Only 1/21 SCCL showed dot-like patterns for pan-CK; 20/21 reacted diffusely. All MCC and 13/22 SCCL displayed CHR-positive cells. All MCC and all SCCL were positive for NSE and negative for S100.. Dot-like positivities for CK-20 or NF are important to prove MCC and to exclude SCCL in clinically and morphologically doubtful cases. Dot-like positivities for pan-CK favor MCC, but do not always exclude SCCL. NSE and CHR are of no value for the differential diagnosis of MCC and SCCL. Melanoma and lymphoma are ruled out by negativity for S100 and pan-CK, respectively.

Topics: Aged; Aged, 80 and over; Carcinoma, Merkel Cell; Carcinoma, Small Cell; Diagnosis, Differential; Eyelid Neoplasms; Female; Humans; Immunoenzyme Techniques; Keratins; Leukemia, Lymphocytic, Chronic, B-Cell; Lung Neoplasms; Middle Aged; Neurofilament Proteins; Phosphopyruvate Hydratase; S100 Proteins

Previously available serum tumor markers had a low clinical value in malignant pleural mesothelioma (MPM). The recently developed tissue polypeptide-specific antigen (TPS) and CYFRA 21-1 assays identify the soluble cytokeratin 18 and 19 fragments, respectively. In MPM these cytokeratins are expressed and may therefore be used as serum tumor markers. In this preliminary study, TPS and CYFRA 21-1 assays were evaluated to determine their potential for management of patients with MPM. Carcinoembryonic antigen (CEA) was evaluated as an additional marker. The study group consisted of 95 patients with benign lung and pleural diseases (BLPD), 14 patients with MPM, 41 patients with adenocarcinoma of lung (AC), and 40 patients with squamous cell carcinoma of lung (SQC). The utilized cutoff points corresponded to a 95% specificity for patients with BLPD. In MPM, TPS showed greater sensitivity (64.3%) than CYFRA 21-1 (50.0%), while CEA showed no sensitivity. In SQC, the marker CYFRA 21-1 had the highest sensitivity (52.5%), whereas in AC the most sensitive marker was CEA (56.1%). Significantly lower levels of CEA were found in MPM compared with BLPD (p < 0.001) or AC and SQC (p < 0.0001). Conversely, TPS levels in MPM were significantly higher than in SQC (p < 0.05). Close correlation of various individual pretreatment marker levels was observed only between TPS and CYFRA 21-1, both in MPM (r = 0.84; p < 0.001) and in non-small cell lung cancer (NSCLC) (r = 0.71; p < 0.001). In serial determinations of the markers during chemotherapy of MPM (n = 10), TPS and CYFRA 21-1 were shown to demonstrate more or less the same pattern of reactivity, although the changes in the TPS levels better reflected the clinical response to therapy. In conclusion, TPS seems to be a more sensitive marker than CYFRA 21-1.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Keratins; Lung Neoplasms; Mesothelioma; Peptides; Pleural Neoplasms

Serum levels of CYFRA 21-1(cytokeratin-19 fragment) and ProGRP (pro-gastrin-releasing peptide), the new prognostic markers of lung cancer, were measured by ELISA (enzyme-linked immunoadsorbent assay) in 27 (for CYFRA 21-1; male 13, female 14; age 54+/-17 years) or 22 (for ProGRP; male 9, female 13; age 59+/-18 years) patients with various serum creatinine levels, 42 haemodialysis (HD) patients (male 24, female 18; age 59+/-14 years) and 30 continuous ambulatory peritoneal dialysis (CAPD) patients (male 18, female 12; age 48+/-9 years). All the patients were without clinical and radiological signs of lung cancer. Positive correlations were found between serum creatinine and serum CYFRA 21-1 and ProGRP levels. Serum levels of CYFRA 21-1 were above the cutoff limit (3.5 ng/mL) in 57% of HD patients (mean 4.07+/-1.56 ng/mL) and in 73% of CAPD patients (mean 4.87+/-1.56 ng/mL). Serum levels of ProGRP were above the cutoff limit (46.0 pg/mL) in 90% of HD patients (mean 107.0+/-59.4 pg/mL) and in 93% of CAPD patients (mean 112.4+/-44.5 pg/mL). Our data indicate that evaluation of renal function is essential when the measurement of these tumor markers is to be applied as one of the diagnostic tools of lung cancer.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Creatinine; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratin-19; Keratins; Kidney Failure, Chronic; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Peptides; Recombinant Proteins

To analyze messenger RNA (mRNA) levels for the alpha, beta, and gamma subunits of the human amiloride-sensitive epithelial Na+ channel (hENaC) in respiratory epithelia, we developed a competitive quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) assay specific for each subunit, using two human respiratory epithelial-cell lines. We next determined the relation between hENaC mRNA levels and the biologic activity of the hENaC in the respiratory epithelium of eight normal men. The electrical potential difference (PD) between the epithelium of the inferior nasal turbinate and the subcutaneous space was measured, using control and amiloride (100 microM) solutions. QRT-PCR measurement of hENaC-subunit mRNAs and epithelial-specific cytokeratin 18 mRNA allowed us to normalize hENaC expression to epithelial-cell RNA. Respective values for alpha, beta, and gamma hENaC mRNA levels in epithelium obtained at the site of maximal PD were 39 +/- 4.0, 7.5 +/- 0.92, and 1.8 +/- 0.25 attomol/fmol cytokeratin mRNA, respectively. Respiratory epithelial PD exhibited a significant negative correlation with gamma hENaC (r2 = 0.72, p < 0.01), tended to increase with increasing alpha hENaC, and was unaffected by beta hENaC mRNA levels. Our results suggest that hENaC activity in vivo is influenced by expression of the gene for gamma hENaC. The assay used in the study provides a useful tool for evaluating Na+-channel expression in clinically relevant patient populations.

Topics: Adenocarcinoma, Papillary; Adult; Amiloride; Carcinoma, Large Cell; Diuretics; Epithelium; Humans; Keratins; Linear Models; Lung Neoplasms; Male; Membrane Potentials; Middle Aged; Nasal Mucosa; Polymerase Chain Reaction; RNA-Directed DNA Polymerase; RNA, Messenger; Skin; Sodium Channels; Tumor Cells, Cultured; Turbinates

The immunohistochemical diagnosis of mesothelioma is commonly made by using a battery of antibodies that reacts with lung adenocarcinomas but not with epithelial mesotheliomas. Only recently have markers that are often expressed in mesotheliomas but not in adenocarcinomas been recognized. Some of these markers, however, require frozen tissue sections, whereas others are not commercially available, or their value remains controversial. In a recent publication, it was suggested that immunostaining for cytokeratin 5/6 could assist in distinguishing epithelial mesothelioma from lung adenocarcinoma. To determine the practical value of cytokeratin 5/6 immunostaining in the diagnosis of mesothelioma, 40 formalin-fixed, paraffin-embedded epithelial pleural mesotheliomas, 30 pulmonary adenocarcinomas, 93 nonpulmonary adenocarcinomas, 15 squamous carcinomas of the lung, 5 large cell undifferentiated carcinomas of the lung, and 12 metastatic transitional cell carcinomas to the lung were stained with the same antibody, which was obtained from a commercial source. Cytokeratin 5/6 reactivity was observed in all 40 mesotheliomas, but there was none in any of the 30 pulmonary adenocarcinomas. Focal or weak reactivity was observed in 14 of 93 nonpulmonary adenocarcinomas (10 of 30 ovarian, 2 of 10 endometrial, 1 of 18 breast, I of 7 thyroid, 0 of 10 kidney, 0 of 10 colonic, and 0 of 8 prostatic). All 15 squamous carcinomas of the lung, 6 of 12 transitional cell carcinomas metastatic to the lung, and 3 of 5 large cell undifferentiated carcinomas of the lung expressed cytokeratin 5/6. It is concluded that cytokeratin 5/6 immunostaining is not only useful in separating epithelial pleural mesotheliomas from pulmonary adenocarcinomas but also can assist in distinguishing epithelial mesotheliomas from nonpulmonary adenocarcinomas metastatic to the pleura.

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Diagnosis, Differential; Epithelial Cells; Evaluation Studies as Topic; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Mesothelioma; Pleural Neoplasms; Retrospective Studies

We describe a 60-year-old woman with leg pain. Although metastatic bone tumor and atypical cells mimicking signet-ring cells in the bone marrow picture were observed, systemic survey revealed no primary lesion. The patient died two months after admission from systemic progress of the disease. Autopsy revealed a small focus of adenocarcinoma within the right upper lobe of the lung and systemic metastases without any particular changes in the gastrointestinal tract. The tumor cells of the lung were diffusely positive for cytokeratin 7, whereas cytokeratin 20 immunoreactivity was weak and focal, and that supported the lung origin of the present tumor. Moreover, the tumor cells in the bone marrow showed a similar pattern in immunoreactivity. These findings suggest that cytokeratin 7 and cytokeratin 20 immunoreactivity is helpful for the premortem diagnosis of the metastatic tumor of unknown origin.

Topics: Biomarkers, Tumor; Bone Marrow; Bone Neoplasms; Carcinoma, Signet Ring Cell; Digestive System; Female; Fibula; Humans; Keratins; Lung; Lung Neoplasms; Middle Aged; Neoplasm Proteins; Neoplasms, Unknown Primary; Organ Specificity; Protein Isoforms; Radionuclide Imaging

Two human cancer cell lines were established from metastatic lesions of an adenocarcinoma (RAL) and a squamous cell (CAEP) carcinoma of the lung. The clinical histories of the patients from whom the cell lines were derived are reported. The lines were maintained in continuous culture with doubling times of 65 (RAL) and 50 (CAEP) hours. The RAL and CAEP cell lines, whose morphology and ultrastructural features are presented, showed extensively rearranged karyotypes with modal number of 85 (RAL) and 98 (CAEP). In particular, chromosome 2 pentasomy and several clonal markers were evident in the RAL cells, whereas a telomeric deletion of chromosome 1, del (1)(q32), was observed in the CAEP cells. The morphologic data were confirmed by high expression of specific antigens for each histotype. A marked positivity of the neuron-specific enolase (NSE) levels was evident by immunoenzymatic assays in the cell lines cytosol with respect to those present in the respective patient's sera. No amplification or rearrangements were evident in the CMYC, LMYC, NMYC, INT-2, ERBB2, HRAS, KRAS, MOS, HST-1 genes by Southern blotting analysis in each cell line. Point mutations in exon 1 of KRAS and in exon 7 of TP53 were evident by polymerase chain reaction (PCR)-DNA sequencing in the RAL cell line, whereas no alterations were present in the HRAS and RB genes. The four genes studied did not show point mutations in the CAEP cell line. The RAL cell line was resistant to all the drugs tested, whereas the CAEP cells were sensitive to vinblastine. These cell lines may represent useful experimental models to investigate lung cancer biology and anticancer drug response.

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Genetic Markers; Humans; Karyotyping; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Tumor Cells, Cultured

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Humans; Keratin-19; Keratins; Lung Neoplasms; Lymphatic Metastasis; Mediastinum

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Mesothelioma; Middle Aged; Neoplastic Cells, Circulating; Pleural Effusion; Pleural Neoplasms

Leukocyte common antigen (CD45/LCA) and keratin expression are generally mutually exclusive in diagnostic surgical pathology. CD45 reactivity is a reliable indicator of the hematolymphoid nature of a tumor, whereas keratin reactivity is typical of epithelial differentiation (carcinomas and some sarcomas). Some lymphomas, however, might lack detectable CD45 expression, whereas occasional ones might express keratins. CD45 immunoreactivity has been considered exquisitely specific for hematopoietic cells. We report three undifferentiated or neuroendocrine carcinomas that showed membrane-associated immunoreactivity for CD45 in addition to showing distinctive keratin cocktail (AE1/AE3) and epithelial membrane antigen reactivity (all cases); also, keratin 7 was demonstrated in one case and keratin 19 in another. Two cases were lymph node metastases of undifferentiated carcinomas, one of them from the lungs and the other of an unknown origin; the former case showed neuroendocrine features. The third case represented a pulmonary large-cell undifferentiated carcinoma. These cases were negative for lineage-specific leukocyte antigens and did not show clonal immunoglobulin heavy-chain gene rearrangements. Electron microscopic studies demonstrated desmosomes and keratin-like tonofilaments in all three cases, thus confirming the epithelial nature of these tumors. The exceptional membrane staining for CD45 seen in these undifferentiated carcinomas might be comparable to experimentally detected incorporation of leukocyte antigens into the cell membranes of nonleukocytic cells in a leukocyte-rich environment. This rare diagnostic pitfall should be considered in the diagnostic surgical pathology of undifferentiated tumors. It is best avoided by employing a panel of leukocyte and epithelial antigens and by use of electron microscopy, if possible.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Neuroendocrine; Desmosomes; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Intermediate Filaments; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Lymphatic Metastasis; Lymphoma; Male; Middle Aged; Neoplasms, Unknown Primary

Cytokeratin 19 is particularly abundant in carcinoma of the lung. The CYFRA 21-1 assay has recently been developed for detection of a cytokeratin 19 fragment in serum. In the current study, the prognostic information provided by the CYFRA 21-1 assay in operable nonsmall cell lung cancer (NSCLC) was analysed. Serum levels of CYFRA 21-1 were measured using an immunoradiometric assay (DiaSorin) in 94 patients with operable NSCLC. Survival and disease-free survival curves related to initial levels of this marker were estimated using the Kaplan-Meier method. Elevated preoperative CYFRA 21-1 levels were identified in 42% of patients with NSCLC. The number of patients with elevated levels of this marker increased with tumour node metastasis (TNM) stage (p=0.02). In univariate analysis elevated levels of CYFRA 21-1 were significantly associated with poor overall survival (p<0.001) and with disease-free survival (p<0.001). The results remained significant when the comparisons were adjusted, using the stratified log-rank test, for patient's TNM stage (p<0.001 for both overall and disease-free survival). Elevated preoperative levels of CYFRA 21-1 decreased the probability of survival or surviving without recurrence 15 months or more after the operation. This was confirmed by the results of the multivariate analysis. In conclusion, CYFRA 21-1 may be an independent prognostic parameter of survival and tumour relapse in nonsmall cell lung cancer and may be useful in identifying resected cancer patients at high risk for treatment failure.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Disease-Free Survival; Female; Humans; Immunoradiometric Assay; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Prognosis; Survival Rate

Cytokeratins are epithelial markers whose expression is not lost during malignant transformation. The level of soluble cytokeratin fragment 19 was measured with an enzyme immunoassay method developed by Boehringer Mannheim (Enzymum-test CYFRA 21-1) in the serum of 200 male and 50 female patients with NSCLC (Non Small Cell Lung Cancer) lung cancer (120 planocellulare and 80 adenocarcinoma in males; 22 planocellulare and 28 adenocarcinoma in females). The comparative group comprised 50 young healthy males and 50 females without any clinical proof for malignancy or any other lung disease. The aim of this investigation was to find out if any possible statistical difference exists in the serum level of CYFRA 21-1 between patients with lung cancer and healthy controls, and also between different types of lung cancers. The mean value of serum CYFRA 21-1 in NSCLC (6.25 ng/ml) was significantly higher than in healthy controls (1.26 ng/ml) (p < 0.001). Sensitivity for CYFRA 21-1 (using 3.3 ng/ml, a cut-off value corresponding to a 98% specificity for healthy controls) in NSCLC was 60.5%. Positive CYFRA 21-1 levels were significantly higher in patient with carcinoma planocellulare (66.2%) than in adenocarcinoma (52.1%). CYFRA 21-1 levels were significantly different between squamous cell carcinoma (6.52 ng/ml) and adenocarcinoma (5.86 ng/ml) (p < 0.05). Our results indicate that CYFRA 21-1 may be a useful tumor marker in NSCLC, especially in carcinoma planocellulare. CYFRA 21-1 may also be useful in identification of the preoperative stages of diseases and the postoperative monitoring of NSCLC.

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Immunoenzyme Techniques; Keratin-19; Keratins; Lung Neoplasms; Male; Sensitivity and Specificity

To investigate the presence of human papillomavirus (HPV) DNA in adenosquamous carcinoma of the lung--which is relatively common in Okinawa but not in mainland Japan--and examine its histological features.. Of 207 cases where primary lung cancers were surgically removed between January 1995 and June 1997 in Okinawa, 23 were adenosquamous carcinoma. HPV was detected by non-isotopic in situ hybridisation (NISH) and polymerase chain reaction (PCR) amplification with primers specific for E6 and E7 regions of the HPV genome. PCR products were analysed by Southern blotting. Immunohistochemical determination of high molecular weight cytokeratin (HMC) and involucrin was also carried out.. 18 cases were positive for HPV DNA by PCR and NISH. HPV types 6, 11, 16, and 18 were found. Seven cases were dual positive for different types of HPV. Using NISH, HPV was also found in the squamous cell components and in neighbouring enlarged adenocarcinoma cells. The HMC and involucrin were demonstrated immunohistochemically in the same areas.. HPV DNA was found in a high proportion (78.3%) of adenosquamous carcinomas in Okinawa, a region where HPV has previously been shown to be prevalent in squamous cell carcinoma of the lung. The adenocarcinoma cells adjacent to the squamous cell carcinoma component were enlarged and positive for HPV, HMC, and involucrin. This is thought to indicate the transition from adenocarcinoma to squamous cell carcinoma.

Topics: Aged; Aged, 80 and over; Base Sequence; Blotting, Southern; Carcinoma, Adenosquamous; DNA, Viral; Humans; In Situ Hybridization; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Papillomaviridae; Polymerase Chain Reaction; Protein Precursors

To set the serum cut-off value of CYFRA21-1 in lung cancer patients in china, and to evaluate the clinical usefulness of CYFRA21-1 as a new tumor marker of lung cancer.. CYFRA21-1 levels were measured with two monoclonal antibodies (Ks19.1 and BM19.21) in the serum of 72 patients with lung cancer, 50 patients with benign lung diseases and 33 post-therapy lung cancer patients. To set the cut-off value, ROC curve was use.. (1) Serum concentrations of CYFRA21-1 in patients with lung cancer were significantly higher than those with benign lung diseases (P < 0.001). CYFRA21-1 levels in patients with squamous cell carcinoma were significantly higher than those in patients with any other subtypes. The more advanced the clinical stages, the higher the levels of CYFRA21-1. (2) If the threshold of CYFRA21-1 was set at 5.5 micrograms/L, its sensitivity and specificity for lung cancer were 63% and 94%, respectively. The sensitivity for squamous cell carcinoma was 78%, which was the highest among all the pathological subtypes. (3) The sensitivity of CYFRA21-1 in non-small cell lung cancer increased with the advancement of clinical stages. (4) CYFRA21-1 could be a good index in monitoring patient's condition and predicting prognosis for lung cancer. (5) When CYFRA21-1 was used as a screening index for lung cancer, the threshold of CYFRA21-1 should be set at the upper left corner of the receiver operating characteristic curve of stage I-II. The value of CYFRA21-1 in early diagnosis for non-small cell lung cancer was found limited.. CYFRA21-1 is a sensitive and specific tumor marker of non-small cell lung cancer, especially of squamous cell carcinoma.

Topics: Adenocarcinoma; Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging

The aim of the present study was to investigate whether trivariate FCM analysis, for the simultaneous detection of two different CK subtypes in combination with DNA content, can be applied to paraffin embedded samples of different types of non-small cell lung cancer in order to evaluate the cell cycle of individual sublines. Single cell suspensions were prepared from 50 microm thick paraffin sections of 22 lung carcinomas by pepsin digestion and immunostained with CK-antibodies which were chosen to distinguish glandular differentiation (adenocarcinomas) and squamous differentiation. There was a good correlation between the immunocytochemical results of the different CK antibodies in tissue sections and in the corresponding single cell suspensions. Gating for CK-positivity revealed a higher S-phase fraction as compared to the ungated cell population. The tumor cells in adenocarcinoma cases were specifically recognized by CK7 antibodies, while well-differentiated squamous cell carcinomas were specifically stained for CK14 and/or CK17. In poorly differentiated squamous cell carcinomas simultaneous expression of CK7 and CK17 was detected in a subpopulation of the tumor cells, next to cells positive for CK7 or CK17 alone. The trivariate FCM analysis allowed the separate estimation of ploidy status and cell cycle parameters in the three different cell populations of these, apparently (phenotypically) heterogeneous, malignancies.

Topics: Adenocarcinoma; Antibodies; Carcinoma, Squamous Cell; Cell Cycle; DNA, Neoplasm; Flow Cytometry; Frozen Sections; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Paraffin Embedding

By means of a mathematical score previously generated by discriminant analysis on 90 lung cancer patients, a new and larger group of 261 subjects [209 with non-small-cell lung cancer (NSCLC) and 52 with small-cell lung cancer (SCLC)] was analysed to confirm the ability of the method to distinguish between these two types of cancers. The score, which included the serum neuron-specific enolase (NSE) and CYFRA-21.1 levels, permitted correct classification of 93% of the patients. When the misclassifications were analysed in detail, the most frequent errors were associated with limited disease SCLC with low NSE levels and with advanced NSCLC with high NSE levels. This demonstrates the importance of the marker in correctly categorizing patients.

Topics: Adult; Aged; Aged, 80 and over; Algorithms; Analysis of Variance; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Diagnosis, Differential; Discriminant Analysis; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Phosphopyruvate Hydratase; Reproducibility of Results

In a previous study, the authors used a variety of anticytokeratin monoclonal antibodies to show that a large proportion of lung tumors cytologically diagnosed as squamous cell carcinoma contain cells expressing simple epithelial cytokeratins, suggesting that these tumors have their origin in adenocarcinoma. These findings raised the possibility that cytokeratin (CK) typing might have a diagnostic capacity not attainable by standard histopathology. The aim of the current study was to assess the value of CK typing for this purpose by determining the correlation between the diagnosis of lung tumors based on CK typing and the survival rate of the patients.. Paraffin embedded tissue sections of 66 nonsmall cell lung carcinoma (NSCLC) specimens were examined. These included 18 adenocarcinomas, 32 squamous cell carcinomas, and 16 undifferentiated carcinomas, all diagnosed surgically and histopathologically, and further classified as either Stage I or II. CK typing was performed using the streptavidin-biotin-peroxidase method, employing the following anti-CK monoclonal antibodies: Ks.B.17 (which reacts with CK 18), A3-3 (which reacts with CK 13), and E5-9 (which reacts with CK 10).. Comparison between the 5-year survival rates (5 ysr) of patients with different NSCLC indicated that all types of Stage II tumors had a much poorer prognosis than Stage I tumors. Differences found in the 5 ysr among patients with different types of Stage I tumors were not statistically significant (adenocarcinomas, 33% 5 ysr; squamous cell carcinomas, 59% 5 ysr; undifferentiated carcinomas, 36% 5 ysr; all diagnosed by conventional histopathology). Similarly, no significant differences were noted in 5 ysr between patients with tumors stained positively or negatively with monoclonal antibodies A3-3 or E5-9 (anti-CK 13 and anti-CK 10, respectively). In contrast, highly significant differences (P = 0.002) were found in the 5 ysr between patients with Stage I tumors positively or negatively stained with monoclonal antibody Ks.B.17 (23% vs. 75% 5 ysr, respectively) regardless of the histologic types of tumors. Especially informative was a combination of immunohistochemical and histologic diagnoses, with best survival rates (87% 5 ysr) in Ks.B.17 negative tumors histologically diagnosed as Stage I squamous cell carcinomas and worst survival rates (14% 5 ysr) in Ks.B.17 positive tumors diagnosed as adenocarcinomas.. The current study showed that CK 18 typing of lung tumors can provide a more accurate diagnosis and therefore facilitate the planning of more suitable therapeutic approaches.

Topics: Adenocarcinoma; Aged; Carcinoma; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Gene Expression Regulation, Neoplastic; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Prognosis; Retrospective Studies; Survival Analysis

The aim of this study was to evaluate the diagnostic value of three tumour markers, squamous cell carcinoma (SCC) antigen, carcinoembryonic antigen (CEA) and CYFRA 21.1, in lung cancer using a Bayesian analysis to obtain the predictive values for different pretest probabilities or prevalences. A cross-sectional study included 94 patients with lung cancer, 40 with benign lung disease, and 40 healthy controls. SCC antigen and CEA were measured in blood samples by microparticle enzyme immunoassay (MEIA), and CYFRA by enzyme-linked immunosorbent assay (ELISA). The results of tumour marker determinations were expressed as percentiles, and showed significantly higher levels in the cancer group than in the two control groups. Taking the 95th percentile of benign lung diseases as the cut-off point (specificity 95%), the following sensitivities were found: SCC 41%, CEA 31% and CYFRA 79%. After a Bayesian analysis, the best results for the three tumour markers were found in prevalences of 30-40%. The highest incremental gain was obtained by CYFRA (at prevalence of 36%, positive and negative predictive value approximately 90%). The three tumour markers were included in a stepwise regression analysis to predict lung cancer, and CYFRA was the only selected variable. We conclude that CYFRA 21.1 may be a useful marker in lung cancer when there is an intermediate pretest probability of disease.

Topics: Adenocarcinoma; Aged; Antigens, Neoplasm; Bayes Theorem; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cross-Sectional Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoenzyme Techniques; Keratin-19; Keratins; Logistic Models; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; ROC Curve; Sensitivity and Specificity; Serpins

A 33-yr-old African-American male with known human immunodeficiency virus (HIV) positivity underwent CT guided fine-needle aspiration biopsy of an anterior mediastinal mass. The aspirate was composed of a dimorphic population of cells that included small mature lymphoid cells and scattered cohesive groups of large epithelial cells in equal numbers. The neoplasm stained strongly for low weight molecular cytokeratin, epithelial membrane antigen (EMA), leukocyte common antigen (LCA), and Leu-7 which was consistent with a diagnosis of thymoma. Subsequent biopsies determined the neoplasm to be a malignant (invasive) thymoma. This case emphasizes the efficacy of FNA biopsy for the evaluation of anterior mediastinal masses in HIV infected individuals. Additionally, the differential cytologic diagnoses for HIV infected individuals for this anatomic site are discussed.

Topics: Adult; CD57 Antigens; Diagnosis, Differential; HIV Seropositivity; Humans; Immunoenzyme Techniques; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Male; Radiography, Thoracic; Thymoma

To elevate the diagnostic value of the serum cytokeratin 19 fragment (CYFRA 21-1) and compare it with carcinoembryonic antigen (CEA) and tissue polypeptide antigen (TPA) in bronchogenic carcinoma, the sera of 161 patients (58 with benign pulmonary disease and 103 with bronchogenic carcinoma) was investigated using immunoradiometric assay. Sensitivities for CYFRA 21-1, CEA and TPA (using 3.5 ng ml-1, 5.0 ng ml-1, 110 U l-1, respectively, cut-off values corresponding to a 95% specificity for benign pulmonary disease) in bronchogenic carcinoma were 64, 47 and 61%, respectively. Positive CYFRA 21-1 levels were identified in 75% of patients with squamous cell carcinoma (n = 36), in 67% with adenocarcinoma (n = 45), in 17% with large cell carcinoma (n = 6), and in 50% with small cell lung cancer (SCLC) (n = 16). However, CYFRA 21-1 levels were not significantly different between squamous cell carcinoma and the other histological types. The sensitivity of the combined measurement of CYFRA 21-1 with any other tumour marker was significantly higher than that of CYFRA 21-1 measurement alone. Elevated CYFRA 21-1 levels were observed in 44% of Stages I and II (n = 18) and 72% of Stage III and IV (n = 69) patients with non-small cell lung cancer (P < 0.05). A significant inter-marker correlation was observed between CYFRA 21-1 and TPA (n = 103, r = 0.448, P < 0.0001). Twenty-one patients were monitored by CYFRA 21-1, and significantly different changes in progressive patients (P = 0.0058) and regressive patients (P = 0.016) were obtained. These results indicate that CYFRA 21-1 may be not only a sensitive tumour marker in the diagnosis of bronchogenic carcinoma, but also a useful marker for the monitoring of bronchogenic carcinoma.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Bronchogenic; Carcinoma, Large Cell; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Immunoradiometric Assay; Keratin-19; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Sensitivity and Specificity; Tissue Polypeptide Antigen

With the exception of two cases, keratin is not expressed in cultured human melanoma cells. Using 2D-PAGE, immunological and electron microscopic analyses, we found keratin subunits in five established cultured cell lines derived from primary, recurrent and metastasized melanomas. The keratin subunits were composed of K1, K5, K10, K14, K15 and K18 in all cell lines examined, together with vimentin. In addition, K8, K16 and K18 expression were demonstrated in recurrent and metastasized cell lines. The results of the present and our previous study [Katagata Y, et al. J Dermatol Sci 1996;13:219-227] indicate that expression of keratin in melanoma cells may be a universal phenomenon. A specific increase in the proportion of K5 among the keratin subunits was suggestive of the nature of melanoma cells. Moreover, we detected two polypeptides that migrated on 2D-PAGE at positions which did not correspond to those of any keratin subunit. The amino acid sequences of these two polypeptides were determined; one was the human ATP synthase alpha-chain but the other did not match any known polypeptide in our homology search.

Topics: Amino Acid Sequence; Antigens, CD; Humans; Keratins; Lung Neoplasms; Lymphatic Metastasis; Melanoma; Membrane Glycoproteins; Molecular Sequence Data; Proton-Translocating ATPases; Recurrence; Sequence Analysis; Tetraspanin 29; Tumor Cells, Cultured

To investigate the usefulness of Cyfra 21-1 as an indicator of therapy effectiveness and prognosis in advanced primary lung cancer, sixty-three patients were selected on the basis of a high Cyfra 21-1 serum level (> 3.3 ng/ml) at the time of diagnosis. Serial assays of Cyfra 21-1 were performed during the first three courses of chemotherapy among 63 patients. The serial-values were analysed according to response to treatment and overall survival. After three courses of chemotherapy, a 70% reduction under the initial marker's value or a return to normal was observed for 36 patients. Twenty-two (61%) of these patients presented an objective response to therapy, making Cyfra 21-1 a moderate indicator in terms of positive predictive value (PPV). However, a significant decrease of Cyfra 21-1 was observed in 88% (sensitivity) of the 25 objective responders. Cyfra 21-1 changes after one course of chemotherapy (61 patients) were not sufficient to predict the future response after three courses (sensitivity 52%, specificity 56%, PPV 45%). Among 30 clinical or radiological relapses, a 10% increase or a return upper reference limit in Cyfra 21-1 level was observed in 18 cases (sensitivity 60%, specificity 100%, PPV 100%). Survival data were available for 61 patients. No significant statistical difference (P > 0.05) was found between survival curves depending on a significant decrease of Cyfra 21-1 after the first course of chemotherapy. We can conclude that the only interest of serial Cyfra 21-1 assays may be the detection of relapse, where one observes a significant decrease of the marker correlated with an objective response to first treatment.

Topics: Adult; Aged; Analysis of Variance; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Disease Progression; Environmental Monitoring; Female; Humans; Keratin-19; Keratins; Longitudinal Studies; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Recurrence; Reference Values; Sensitivity and Specificity; Survival Rate

The diagnostic value of Cyfra 21-1 in non-small lung cancer (NSCLC) has been established, but few studies have focused on its prognostic value. The aim of this study was to compare that of carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 19-9, CA 125, neuron-specific enolase and squamous cell carcinoma antigen. 116 patients with unresectable (n = 88) or resectable (n = 28) NSCLC were prospectively monitored from diagnosis, for a median of 14.4 months. All patients underwent tumour-marker determinations before treatment, then every 3 months. Their diagnostic value was studied using ROC (receiver operating characteristic) curves, based on control measure in 23 patients with benign lung diseases. The prognostic analysis was based on overall survival as the main endpoint. The diagnostic value of Cyfra 21-1 was confirmed, with a sensitivity of 54% and a specificity of 96% at a cut-off value of 3.3 ng/ml. At diagnosis, in the 88 non-surgical NSCLC, besides the presence of metastases (P = 0.017), Cyfra 21-1 (P = 0.017) and CA 125 (P = 0.03) were related to outcome. Elevated levels of Cyfra 21-1 at any time during the disease course was selected by multivariate analysis as additional predictors of poor survival.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Disease-Free Survival; Female; Follow-Up Studies; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Prognosis; Prospective Studies; Sensitivity and Specificity; Survival Rate

Cytokeratins form part of the cytoskeleton of both normal and malignant epithelium. A novel tumor marker measuring cytokeratin 19 (CK-19) fragment has been introduced and proven to be suitable for monitoring therapy and following cases of non-small cell lung cancer, squamous cell lung cancer in particular. However, whether the serum level of CK-19 fragment reflects the number of proliferating tumor mass remains unknown. We studied the CK-19 fragment produced by two human squamous cell lung cancer cell lines. In Western blotting analysis, culture supernatants of both cell lines displayed bands of 37 and 40 kDa, which represented the CK-19 fragment and the intact CK-19, respectively. Gel filtration demonstrated that a part of soluble CK-19 was released as a large complex form in culture supernatants. The level of CK-19 fragment in culture supernatants increased during the exponential growth phase. CK-19 level decreased by an addition of a cytotoxic agent to non-significant level though the transient release of CK-19 fragment occurred during the first 2 days. After all, soluble CK-19 fragments were detected in culture supernatants of human lung cancer cell lines and its level reflected proliferating cancer cells though it was not determined whether CK-19 fragments were released directly from live cells.

Topics: Carcinoma, Squamous Cell; Cell Division; Cell Survival; Humans; Keratins; Lung Neoplasms; Molecular Weight; Peptide Fragments; Tumor Cells, Cultured

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Humans; Keratin-19; Keratins; Lung Neoplasms; Neoplasm Staging; Phosphopyruvate Hydratase

This study was designed to evaluate the incidence and clinical implications of detection of micrometastatic cancer cells in bone marrow aspirates of patients with operable non-small cell lung cancer. The relationship between micrometastatic cells and p53 overexpression in the primary tumor was also assessed.. Thirty-nine patients with stages I through III non-small cell lung cancer who underwent curative resection were entered into this study. Immunohistochemistry with monoclonal antibody to cytokeratin 18 was used to detect tumor cells in bone marrow. Immunostaining of p53 protein in the corresponding primary tumors was also done.. Cytokeratin 18-positive cells were detected in 15 (39%) of the 39 patients. Overexpression of p53 was associated with positivity of the tumor cells in the bone marrow at borderline significance (14/29 versus 1/10; p = 0.0574). The patients with cytokeratin 18-positive cells in bone marrow demonstrated a significantly earlier recurrence than those without such cells (p = 0.0083, log-rank test).. Micrometastatic cancer cells were frequently present in bone marrow of patients with operable non-small cell lung cancer and may be a significant predictor of early recurrence. Further evaluation of this method may be useful in identifying patients with non-small cell lung cancer who are most likely to benefit from adjuvant chemotherapy.

Topics: Aged; Biomarkers, Tumor; Bone Marrow Neoplasms; Carcinoma, Non-Small-Cell Lung; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Prognosis; Tumor Suppressor Protein p53

We report on a rare tumor of the lung characterized by its morphologic hepatoid features and alpha-fetoprotein production. This unusual neoplasm arose in the left lung of a 36-year-old man in whom clinical and radiologic examinations did not reveal any other tumor. The serum level of alpha-fetoprotein was measured at 6,090 ng/mL and was parallel to the evolution of the tumor. Despite treatment, the patient died 7 months after the diagnosis. The microscopic appearance of the tumor was the same as observed in hepatocarcinoma and hepatoid adenocarcinoma of the ovary or the stomach, with a tubular, papillary, or trabecular pattern. Periodic acid-Schiff-positive hyaline globules were numerous, and tumor cells showed immunohistologic positivity for alpha-fetoprotein and carcinoembryonic antigen. This lung adenocarcinoma was first described by Ishikura et al. in 1990 and was named hepatoid lung adenocarcinoma. Like the rare hepatoid carcinoma of the gallbladder, the pancreas, the ampulla of Vater, the renal pelvis, and the bladder, the exact histogenesis and the prognosis of this type of lung tumor are not yet known.

Topics: Adenocarcinoma; Adult; alpha-Fetoproteins; Carcinoma, Hepatocellular; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Magnetic Resonance Imaging; Male; Mucin-1; Prognosis; Time Factors

During apoptosis, one of the first membrane changes that can be detected is exposure of phosphatidylserine residues at the outer plasma membrane leaflet, while early apoptosis is also accompanied by changes in the cytoskeletal organization. In this study we investigated the relationship between these two phenomena during olomoucine- and roscovitin-induced apoptosis in human lung cancer and neuroblastoma cell lines. Loss of membrane asymmetry was detected by biotin-labeled or FITC-labeled annexin V binding to negatively charged phosphatidylserine, while cytoskeletal components were visualized by immunocytochemistry. The apoptotic, annexin V-positive, cells were analyzed by flow cytometry, confocal scanning laser microscopy, and Western blotting. We report that cytokeratin and vimentin aggregation in early apoptosis occurs simultaneously with phosphatidylserine exposure and chromatin condensation. In contrast to these intermediate filament proteins, which were disassembled and proteolytically cleaved in early apoptosis, microfilaments and microtubuli were not proteolytically degraded but were found to be present as aggregated filaments in the apoptotic bodies. We also show that loss of membrane asymmetry and cytokeratin aggregation are independent processes, since N-ethylmaleimide-induced phosphatidylserine exposure does not cause cytokeratin disassembly. Vice versa, phorbol 12-myristate 13-acetate-induced cytokeratin filament aggregation does not result in phosphatidylserine exposure.

Topics: Actins; Annexin A5; Apoptosis; Carcinoma, Non-Small-Cell Lung; Cell Membrane; Chromatin; Cytoskeleton; Ethylmaleimide; Humans; Keratins; Kinetin; Lung Neoplasms; Neuroblastoma; Phosphatidylserines; Purines; Roscovitine; Tetradecanoylphorbol Acetate; Tubulin; Tumor Cells, Cultured; Vimentin

To better understand the mechanism(s) underlying lung cancer invasion and metastasis, a Transwell invasion chamber was used to select progressively more invasive cancer cell populations from a clonal cell line of human lung adenocarcinoma, CL1. Five sublines with progressive invasiveness, designated CL1-1, CL1-2, CL1-3, CL1-4, and CL1-5, were obtained through this in vitro selection process. Their invasive abilities through basement membrane matrix showed a 4- to 6-fold increase over that of the parental cells. Moreover, the sublines manifested an increase in their colony-forming ability on soft agar, tumorigenicity, and metastatic potency in severe combined immunodeficiency (SCID) mice. Examining the phenotypes of the cell lines revealed increased expression of 92 kD gelatinase and an increase in the cell population stained with anti-keratin-8 and -18 antibodies. Clonal isolation of anti-keratin-18-antibody-positive and -negative cell populations demonstrated a correlated enhancement of the invasiveness of these cells and their expression of keratin-18. These results support the notion that the metastatic behavior of lung cancer cells can be characterized with this in vitro system, and that the properties of these progressively invasive cancer cells can be clonally studied.

Topics: Adenocarcinoma; Carcinogenicity Tests; Cell Culture Techniques; Gelatinases; Humans; Keratins; Lung Neoplasms; Male; Microscopy, Electron; Middle Aged; Neoplasm Invasiveness; Tumor Cells, Cultured

Topics: Blood Cells; Humans; Keratins; Lung Neoplasms; Medical Laboratory Science; Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Transcription, Genetic

Conflicting results have been reported on the use of cytokeratin-19 (CK-19) in the detection of tumor cells in the peripheral blood of patients with solid tumors. We investigated the expression of CK-19 in lung cancer cell lines and in human lung tumor samples using a nested reverse transcriptase (RT)-PCR to determine the sensitivity and specificity of this method. In addition, blood samples of lung cancer patients and healthy controls were analyzed for the presence of CK-19 transcripts. Amplification products were visualized by ethidium bromide staining and radioactive hybridization with a CK-19-specific probe. Application of a previously described nested RT-PCR for the detection of CK-19 resulted in amplification of the processed pseudogene. Therefore, a more stringent RT-PCR was developed by increasing the annealing temperature. RT-PCR amplification products for CK-19 were detected in 38 of 41 lung cancer cell lines. The three negative cell lines were all variant small-cell lung cancer cell lines. Concordant results were observed between CK-19 detection by immunohistochemistry and by RT-PCR. In serial RNA dilution experiments, CK-19 transcripts could be detected in 18 to 80 pg of total cellular RNA in three cell lines and in 60 ng total RNA in one cell line. The nested RT-PCR had the sensitivity of detecting 50 tumor cells in 10(6) peripheral blood mononuclear cells (PBMNC), and CK-19 transcripts were randomly detected in normal PBMNC. This study shows the necessity in processing parallel samples without reverse transcriptase enzyme to avoid amplification of pseudogenes. A serious problem in the detection of tissue-specific transcripts in PBMNC is the detection of illegitimate transcription levels. In conclusion, although CK-19 may be a useful marker for the detection of lung cancer cells, its application for the detection of circulating tumor cells is not recommended.

Topics: Blood Cells; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Polymerase Chain Reaction; Reference Values; RNA, Messenger; Sensitivity and Specificity; Transcription, Genetic; Tumor Cells, Cultured

Although vascular invasion is common in many malignant tumors, disseminated intravascular anaplastic neoplasms with occult primary tumor are rare occurrences. Intravascular malignant lymphoma, also called angiotropic lymphoma, is a rare variant of large cell lymphoma predominantly involving vessels in multiple organs, and usually without significant nodal involvement. Although initially misinterpreted as an endothelial neoplasm-angioendotheliomatosis-immunohistochemical studies subsequently proved it to represent a peculiar form of malignant lymphoma. In this report, we describe two patients with extensive intravascular dissemination of angiosarcoma initially without clinically obvious primary tumor. These may be interpreted as examples of true angioendotheliomatosis. In each case the immunohistochemical studies ruled out the most common intravascular malignant neoplasms. The diagnosis of intravascular angiosarcoma was confirmed by the immunoreactivity of the tumor cells to several markers of endothelial lineage in both cases. Thus, angiosarcoma may present with intravascular dissemination and occult primary tumor and closely resemble metastatic carcinoma, melanoma, or angiotropic lymphoma. Immunohistochemical studies are crucial in ruling out these possibilities and in confirming the endothelial origin of the neoplastic cells.

Topics: Adult; Aged; Biomarkers; Biopsy; Diagnosis, Differential; Fatal Outcome; Female; Gallbladder Neoplasms; Hemangiosarcoma; Humans; Immunohistochemistry; Keratins; Leukocyte Common Antigens; Liver Neoplasms; Lung Neoplasms; Platelet Endothelial Cell Adhesion Molecule-1; Vascular Neoplasms; von Willebrand Factor

Cytokeratins (CK) are one of the main families of intermediate filaments which make up the cytoskeleton. CK19 is strongly expressed by normal simple bronchial and respiratory epithelium as well as by their malignant counterpart. Although CK19 is a part of the cytoskeleton, a soluble fragment of this polypeptide can be released and assayed in the blood as CYFRA 21-1, new sensitive and valuable marker of non small cell lung cancer. In some cases, however, discrepancies between the serum level of CYFRA 21-1 and presence of tumor and its histological type have been observed. We studied immunohistochemically tissue localization of CK19 in tumors and non invaded lung parenchyma in a series of 34 patients surgically treated due to lung cancer. CK 19 was detected in cancer cells as well as in non neoplastic epithelium covering bronchial tree and alveolar surfaces. We found a different expression of CK19 in different histological type of tumors. The most intensive expression revealed squamous cell carcinomas and adenocarcinomas. Small cell cancer revealed poor expression of CK19. In non invaded parts of the resected lungs we found the strong expression of CK19 in the cytoplasm of regenerative II type pneumocytes occurring in large quantity in the cases of interstitial lung fibrosis concomitant with some tumors. We suggest it may be a cause of unexpectedly elevated serum levels of CYFRA 19-21 in some not oncological patients or patients with small cell lung cancer.

Topics: Adenocarcinoma; Adult; Aged; Antibodies, Monoclonal; Biomarkers, Tumor; Bronchi; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cytoplasm; Epithelium; Female; Humans; Infant; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Pulmonary Alveoli; Pulmonary Fibrosis

This retrospective study was designed to detect occult micrometastases in the lymph nodes with the use of monoclonal anti-cytokeratin reagent, which is specific for epithelial cells but not for lymphocytes or plasmacytes, as well as to assess the relationship between the presence of occult micrometastases in the lymph nodes and an early relapse in patients with stage I non-small-cell lung cancer.. The paraffin-embedded sections of 973 regional lymph nodes from 44 patients with stage I non-small-cell lung cancer were studied. We used CAM-5.2 as the primary monoclonal anti-cytokeratin reagent and an indirect staining technique with the streptavidin-biotin-peroxidase complex method.. We identified cytokeratin-positive cells in 31 (70.5%) of 44 patients and in 91 (9.4%) of the 973 lymph nodes. Of these 31 patients with cytokeratin-positive cells, 19 and 12 were restaged as having N1 and N2 disease, respectively. Thirteen patients had recurrent disease at 17 sites during the follow-up. Two of these recurrences were in the mediastinal nodes and the other 15 occurred at distant organs. Twelve of the 13 patients had micrometastatic disease in the regional lymph nodes. Disease-free survival duration was significantly shorter in the patients with micrometastases in the mediastinal lymph nodes than in patients with node-negative disease (p = 0.004). The independence of this prognostic significance was demonstrated by a multivariate analysis.. These findings indicate that the detection of occult micrometastases in the mediastinal lymph nodes with monoclonal antibodies to cytokeratin can thus be used to predict an early relapse in patients with stage I non-small-cell lung cancer.

Topics: Aged; Antibodies, Monoclonal; Carcinoma, Non-Small-Cell Lung; Case-Control Studies; Disease-Free Survival; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Mediastinum; Middle Aged; Neoplasm Staging; Retrospective Studies; Time Factors

We describe an unusual case of a basaloid squamous-cell carcinoma (BSCC) of the tonsil in a 56-yr-old man that metastasized to a primary renal-cell carcinoma (RCC) and the lung. The diagnosis of the second primary, the RCC, was based on fine-needle aspiration (FNA) cytology. A subsequent nephrectomy specimen revealed BSCC metastatic to RCC, clear-cell type. Retrospective analysis of the FNA of the renal mass revealed classic RCC morphology and, in addition, another cytologically distinctive pattern characterized by occasional sheets of cohesive neoplastic cells with hyperchromatic nuclei and nuclear molding representative of BSCC. The cytologic features of a subsequent FNA of the lung were characteristic of metastatic BSCC. Our retrospective analysis of cytologic material from the renal mass underscores the importance of raising the possibility of tumor-to-tumor metastasis when distinctly different morphologic features are seen in an otherwise typical cytology of a neoplasm in patients with an already known or suspected second primary. To our knowledge, this case report is the first one documenting metastasis of BSCC to RCC.

Topics: Biopsy, Needle; Carcinoma, Basosquamous; Carcinoma, Renal Cell; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasms, Second Primary; Tonsillar Neoplasms

The immunohistochemical diagnosis between epithelial mesothelioma and adenocarcinoma is currently based on the use of a panel of antibodies to adenocarcinoma-associated antigens and a few antibodies to mesothelial-associated antigens. Since the introduction of epitope retrieval methods, the sensitivity of many antibodies has been enhanced. Thus, a reevaluation of the mesothelioma/adenocarcinoma diagnostic panel becomes necessary. We studied 268 paraffin-embedded formalin-fixed tumor samples that included 57 epithelial mesotheliomas and 211 adenocarcinomas of various origins, comparing an extensive antibody panel with and without heat-induced epitope retrieval (HIER). Marked increase in the sensitivity of several antibodies, with no loss of specificity, was found when HIER was used. After statistical analysis, the antibodies to the epithelial glycoproteins carcinoembryonic antigen, BerEp4, and Bg8 emerged as the best discriminators between adenocarcinoma and epithelial mesothelioma within the entire panel. The mesothelium-associated antibodies, HBME-1, calretinin, and thrombomodulin were less sensitive and less specific than the former, although they were found to be useful on certain cases. Antibodies to cytokeratins and vimentin, although of minor diagnostic value in this context, may be helpful to evaluate the quality of antigen preservation. This study confirms the value of immunohistochemistry to accurately distinguish mesothelioma from adenocarcinoma when an antibody panel approach is used. The addition of heat-induced epitope retrieval methods increases the effectiveness of the procedure and is recommended for most of the antibody panel members.

Topics: Adenocarcinoma; Biomarkers, Tumor; Breast Neoplasms; Calbindin 2; Carcinoembryonic Antigen; Colorectal Neoplasms; Decision Trees; Diagnosis, Differential; Epitopes; Female; Hot Temperature; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mesothelioma; Ovarian Neoplasms; Peritoneal Neoplasms; Pleural Neoplasms; Retrospective Studies; S100 Calcium Binding Protein G; Sensitivity and Specificity; Thrombomodulin; Vimentin

Keratin 8 and 18 are commonly used as tumorigenic markers for various types of carcinomas. They are known to be involved in cell migration, cell invasiveness, plasminogen activity and drug and radiation resistance. To ascertain a potential function for simple epithelium keratins in mammary adenocarcinoma in vivo, keratin-8-deficient mice (mK8) were mated with transgenic mice carrying the middle T oncogene driven by the MMTV promoter. The resulting mK8 knockout and control progeny carrying the middle T transgene developed mammary gland tumours with the same incidence. However, the onset of palpable mammary gland tumours occurred earlier in mK8 mutant than in control mice. This effect was prominent in males where the onset in control animals is delayed overall, because of the lower hormonal inducibility of the MMTV promoter. Metastatic foci were observed in the lungs of all females and of a few males, independently of the genotype. Histological analysis revealed no morphological differences of the tumorigenic cells in primary tumours nor in metastatic foci. As expected, keratin 8 was absent in the mK8 tumours. Keratin 7 (mK7), keratin 18 (mK18) and keratin 19 (mK19) protein were observed in both primary and metastatic foci. These results constitute the first in vivo analysis of the role of simple epithelium keratins in mammary carcinogenesis. It demonstrates that the latency, but not the incidence nor the morphological features, of PyV middle T-induced mammary gland tumours is affected by keratin 8 deficiency.

Topics: Age Factors; Animals; Antigens, Polyomavirus Transforming; Biomarkers, Tumor; Female; Heterozygote; Keratins; Lung; Lung Neoplasms; Male; Mammary Glands, Animal; Mammary Neoplasms, Animal; Mammary Tumor Virus, Mouse; Mice; Mice, Knockout; Mice, Transgenic; Mitotic Index; Sex Characteristics

In this study we looked at what useful information cytokeratin fragment detected by antibodies BM 19-21 and KS 19-1 (CYFRA 21.1), carcinoembryonic antigen (CEA), neurone-specific enolase (NSE), tissue polypeptide specific antigen (TPS), and tissue polypeptide antigen (TPA) gave when measured prospectively. All patients who were suspected of having lung cancer and who underwent diagnostic bronchoscopy in this hospital between July 1994 and May 1995 were included in the study. Of 184 patients, 87 were subsequently found to have intrathoracic malignancy, 93 were found to have benign lung disease and four were lost to follow-up. CYFRA 21.1 was the most efficient marker in differentiating benign from malignant disease, with a sensitivity of 54% and a positive predictive value of 96%. Thirty seven patients who had a negative bronchoscopy subsequently turned out to have malignant disease. Either CYFRA 21.1 or CEA was elevated in 26 (70%) of such patients. Multivariate analysis showed that only CYFRA 21.1 and CEA contributed significantly to the discriminatory power of the data. We conclude that measurement of cytokeratin fragment detected by antibodies BM 19-21 and KS 19-1 and carcinoembryonic antigen at the time of bronchoscopy significantly increased the diagnostic yield in this population and was especially useful in those patients in whom tumour biopsy was not possible at bronchoscopy.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Diagnosis, Differential; Humans; Keratin-19; Keratins; Lung Diseases; Lung Neoplasms; Peptides; Phosphopyruvate Hydratase; Predictive Value of Tests; Sensitivity and Specificity; Tissue Polypeptide Antigen

There is controversy about the prognostic significance of occult lymph node metastases detected by immunohistochemistry with the anti-cytokeratin antibody CAM 5.2. The aim of this study was to characterize occult lymph node metastases in colorectal carcinomas that might be associated with a higher risk of recurrence. Three hundred fifty-eight lymph nodes from 10 recurrent and 9 nonrecurrent cases of colorectal carcinoma were examined. All these patients had been reported originally as having no lymph node metastases by routine hematoxylin and eosin staining. Three 10-micron sections or ten 3-micron sections (30-micron total thickness) from each lymph node were stained with CAM 5.2 and examined for the presence of occult lymph node metastases. Occult metastases were detected in 67 of 175 lymph nodes from the recurrent cases, and in 23 of 183 lymph nodes from the nonrecurrent cases. The frequency of positive nodes was significantly higher in the recurrent cases. The recurrent cases had metastases in nodes more distant from the main tumor than did the nonrecurrent cases. Detection of occult lymph node metastases with cytokeratin immunohistochemistry may make it possible to identify patients with a higher risk of recurrence after the removal of a primary colorectal tumor.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Keratins; Liver Neoplasms; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Recurrence, Local; Platelet Endothelial Cell Adhesion Molecule-1; Prognosis

We evaluated the correlation between serum cytokeratin 19 fragment (CYFRA 21-1) and tissue polypeptide antigen (TPA) levels in 57 non-small cell lung cancer patients. There was a significant correlation between serum CYFRA 21-1 and TPA levels for each clinical stage and TNM (T, primary tumor; N, regional lymph node involvement; M, occurrence of distant metastasis) subcategory (range of r-value = 0.809-0.998, P < 0.01). High correlations between serum CYFRA 21-1 and TPA levels were found in eight patients both before and after the surgery, in 22 patients before and after chemotherapy and in another 27 patients who could not complete the scheduled chemotherapy (range of r-value = 0.856-0.998, P < 0.0001). However the positive rate of CYFRA 21-1 was higher than that of TPA (61% vs. 53%, P < 0.05). CYFRA 21-1 would yield better diagnostic results for non-small cell lung cancers than TPA, though these tumor markers are both cytokeratin-associated tumor markers.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Neoplasm Staging; Tissue Polypeptide Antigen

Processed pseudogenes of residual contaminating genomic DNA interfere with a sensitive detection of cytokeratin 18 (CK18) mRNA by reverse transcription and polymerase chain reaction (RT-PCR). This may cause false-positive results when CK18 mRNA is used as a marker for ectopic tumor cells in specimens from cancer patients. To establish a sensitive CK18 RT-PCR by excluding the amplification of processed pseudogenes, the following strategy was chosen: (a) CK18 pseudogene sequences were cloned from genomic DNA by PCR; (b) cDNA-specific primers were designed on the basis of mismatches between pseudogenes and cDNA; (c) PCR conditions were adjusted to reach maximum sensitivity and specificity. Epithelial cells (1-10) could be detected in 1 mL of blood. Among the numerous CK18 genes homologous to the transcribed gene, at least two different processed pseudogenes exist that are highly homologous to each other and to the exons of the transcribed CK18 gene.

Topics: Biomarkers, Tumor; Bone Marrow; Cardia; DNA; DNA Primers; DNA, Complementary; Esophageal Neoplasms; Humans; Keratins; Lung Neoplasms; Molecular Sequence Data; Polymerase Chain Reaction; Pseudogenes; RNA, Messenger; RNA, Neoplasm; Sensitivity and Specificity; Sequence Homology, Nucleic Acid; Stomach Neoplasms

Immunohistochemical studies using epithelial markers have recently been published which identified micrometastases in lymph nodes that had not been found on routine pathological assessment, therefore increasing the accuracy of staging of non-small cell lung cancers. The presence of these micrometastases was associated with reduced survival. We have therefore performed a retrospective immunohistochemical study on all the lymphoid tissue from five lymph node stations (2 hilar, 3 mediastinal) from 49 patients with T1-2, N0 disease. Before immunohistochemistry was undertaken, all slides were reviewed, with the lymph nodes confirmed as negative. In total, 1447 lymph node slices (average 30 per case, 5.9 per lymph node station) were examined, these figures reflecting sectioning of lymph nodes at approximately 3 mm intervals before processing. MNF116, a broad spectrum anti-keratin antibody was then used to look for occult metastases, with adjacent serial sections being examined to ensure that any positively staining cells were detected solely by immunohistochemistry and not through deeper sectioning. In five cases, lymph nodes contained positively staining cells. Two cases proved to be false positives, further immunohistochemistry identifying the cells as benign mesothelial inclusions. In the remaining three cases, positive staining correlated with tumour cells in the adjacent serial sections. Follow-up on 46 of 49 patients revealed recurrence in 27% (actual survival 68%); however all three cases containing tumour cells on immunohistochemistry were free from recurrence. These results suggest that the use of immunohistochemistry adds little useful information above that of thorough routine examination of lymph nodes. They also document that benign mesothelial inclusions within lymph nodes are more frequent than previously reported.

Topics: Antibodies; Carcinoma, Non-Small-Cell Lung; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Neoplasm Staging; Retrospective Studies; Staining and Labeling

A case of epithelioid sarcoma in the tongue is reported. The patient, a 35 year old woman, presented with a non-ulcerated painful lesion of the tongue. Microscopically, the tumour was characterised by multiple coalescent nodules with central geographic necrosis infiltrating the lingual muscle. The tumour cells were epithelioid with abundant eosinophilic cytoplasm and atypical nuclei. Immunohistochemically, the tumour cells stained for vimentin, keratin, and epithelial membrane antigen. These morphological and immunohistochemical appearances led to the diagnosis of epithelioid sarcoma of the tongue. Seven years later, the patient died with metastatic dissemination to the scalp, lungs, and brain. No case of epithelioid sarcoma arising in the tongue has been described previously.

Topics: Adult; Brain Neoplasms; Fatal Outcome; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Sarcoma; Scalp; Skin Neoplasms; Tongue Neoplasms

Histologic, immunohistochemical and ultrastructural features of a case of carcinosarcoma of the lung are described. The biphasic pattern of this neoplasm is characterized by the presence of a carcinomatous component that corresponds to an adenosquamous carcinoma, and a sarcomatous component with rhabdomyoblastic differentiation. Since the biphasic sarcomatoid carcinoma has an aggressive clinical behaviour, immunohistochemical expression of prognostic markers, such as Ki-67 and p53 is evaluated to individuate differences between the carcinomatous and the sarcomatous components of the tumor. The higher p53 expression and Ki-67 positivity in the former, suggests that the carcinomatous component probably represents the more aggressive portion of the tumor. Moreover, P53 expression is nuclear in both carcinomatous and sarcomatous areas, thus it is likely that the biphasic sarcomatoid carcinoma of the lung is monoclonal in origin.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinosarcoma; Cell Differentiation; Cell Nucleus; Desmosomes; Humans; Immunoenzyme Techniques; Keratins; Ki-67 Antigen; Lung Neoplasms; Male; Neoplasm Invasiveness; Neoplasm Proteins; Neoplasms, Muscle Tissue; Tumor Suppressor Protein p53

Cloned v-raf, v-raf/v-myc, and spontaneously transformed rat liver epithelial (RLE) cell lines were examined for meastatic capability in nude mice, using the LacZ gene as a marker for quantitation of micrometastases. Six cloned lines (R3611-T lines) derived from nude mouse xenografts of the v-raf transformed R3611-3 cells displayed variable metastatic capabilities. Three of six subcutaneously inoculated R3611-TlacZ lines produced spontaneous lung metastasis in nude mice. One of the lines, R3611-T2lacZ was highly efficient at metastatic conversion and produced more lung colonies than a faster growing v-raf/v-myc-transformed RJ2-14lacZ line. The spontaneously transformed RLElacZ line (C4T) was nonmetastatic, although it produced larger subcutaneous tumors than the metastatic R3611-T2lacZ line. Metastatic conversion correlated with upregulation of urokinase-type plasminogen activator receptor RNA expression and downregulation of plasminogen activator inhibitor-1, collagen alpha1 (I), and cytokeratin 14 (K14) RNA expression. These findings indicate that proteolytic activities associated with plasminogen activation play a role in the metastatic development in this model. Decreased production of extracellular proteins and cytoskeletal changes associated with lack of K14 expression are also likely to have contributed to the metastatic conversion of the RLE transformants.

Topics: Animals; Cell Transformation, Neoplastic; Clone Cells; Collagen; Down-Regulation; Epithelial Cells; Gene Expression Regulation, Neoplastic; Genes, myc; Keratin-14; Keratins; Liver; Liver Neoplasms, Experimental; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Oncogene Proteins v-raf; Phenotype; Plasminogen Activator Inhibitor 1; Rats; Rats, Inbred F344; Receptors, Cell Surface; Receptors, Urokinase Plasminogen Activator; Retroviridae Proteins, Oncogenic; RNA, Messenger

The monitoring of serum concentrations of Cyfra 21-1, tumor polypeptide antigen (TPA), and tissue polypeptide specific antigen (TPS) has been demonstrated to be useful in the clinical treatment of patients with lung cancer. This study was planned to evaluate the clinical usefulness of the assay of these tumor markers on bronchial washing (BW) fluid and to compare it with serum assay in patients with neoplastic and nonneoplastic disease.. Serum and BW fluid levels of Cyfra 21-1, TPA, and TPS were measured in 40 subjects (10 control subjects, 11 with chronic bronchitis, 10 with squamous cell lung cancer, and 9 with nonsquamous cell lung cancer) undergoing diagnostic bronchoscopy. BW was performed using 25 mL of pyrogen-free saline solution instilled through the working channel of the bronchoscope, and successively aspirated. The quantity of the fluid recovered was measured and used for the assay of albumin, Cyfra 21-1, TPA, and TPS.. Mean BW concentrations of Cyfra 21-1, TPA, and TPS concentrations were significantly higher than serum concentrations (p < 0.01). Serum Cyfra 21-1, TPA, and TPS concentrations were significantly lower in controls and in those with chronic bronchitis than in patients with epidermoid and nonepidermoid carcinoma (p < 0.01). No difference in serum concentrations of the three markers was observed between controls and patients with chronic bronchitis. On the contrary, BW Cyfra 21-1 and TPA concentrations were significantly higher in those with chronic bronchitis and in cancer patients than in controls (p < 0.01), whereas they did not differ between patients with chronic bronchitis and cancer patients. No significant difference in BW TPS concentration was observed among the four groups. Sensitivity and specificity of the BW markers in diagnosing lung cancer were as follows: 68.4% and 61.9% for Cyfra 21-1; 68.4% and 66.6% for TPA; and 57.9% and 66.6% for TPS.. BW fluid concentrations of Cyfra 21-1 and TPA are increased in patients with chronic bronchitis and in patients with lung cancer. Being unable to distinguish malignant from nonmalignant inflammatory conditions, the measurement of airway concentrations of such markers has a too-low specificity to be considered useful in diagnosing malignant abnormalities of the lung.

Topics: Adult; Aged; Albumins; Antigens; Antigens, Neoplasm; Biomarkers, Tumor; Bronchitis; Bronchoalveolar Lavage Fluid; Bronchoscopy; Carcinoma; Carcinoma, Squamous Cell; Chronic Disease; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Peptides; Sensitivity and Specificity; Tissue Polypeptide Antigen

Cytokeratins are the intermediate filaments of the cytoskeletal protein located in normal epithelia, tumor, and cultured cells. Recently, a fragment of cytokeratin subunit 19, referred to as CYFRA 21-1, detected in the serum of patients with nonsmall cell lung cancer, has been reported as a new tumor marker. This article reports the results of a study of serum fragment CYFRA 21-1, measured by immunoradiometric assay, as a marker of lung cancer.. One hundred fourteen patients with primary lung cancer, 6 patients with malignant solid tumor, 116 patients with a variety of benign diseases, and 29 normal individuals were entered into the study. Serum CYFRA 21-1 levels were obtained by means of immunoradiometric assay using the CYFRA 21-1 EIA (enzyme immunoassay) kit. In addition, we studied other tumor markers, including carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), and neuron specific enolase (NSE), as a means of lung cancer diagnosis.. The diagnostic accuracy and sensitivity of serum CYFRA 21-1 for the detection of lung cancer were highest among the four markers. The serum CYFRA 21-1 levels were most highly elevated in lung carcinoma patients (in particular UICC Stage IV patients) across different histologic types and attained 85.1% sensitivity when using a threshold of 3.5 ng/mL. The diagnostic sensitivity for detecting lung carcinoma was substantially enhanced by means of combined assays of CYFRA 21-1 with CEA overall for lung cancer, with SCC for squamous cell carcinoma, and with CEA for adenocarcinoma.. These findings suggest that serum assays of CYFRA 21-1 are clinically useful for the diagnosis of lung carcinoma.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Female; Humans; Immunoradiometric Assay; Keratins; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Phosphopyruvate Hydratase; Sensitivity and Specificity; Serpins

Topics: Biomarkers; Carcinoma, Non-Small-Cell Lung; Humans; Keratins; Lung Diseases; Lung Neoplasms

The CYFRA 21-1, a newly developed sandwich enzyme-linked immunosorbent assay (ELISA), was used to measure soluble cytokeratin 19 fragment in serum that is expressed in simple epithelium and its malignant counterpart. The present study was designed to investigate whether CYFRA 21-1 is a sensitive and specific tumor marker for non-small cell lung cancer.. CYFRA 21-1 assay, using two specific monoclonal antibodies (KS 19.1 and BM 19.21) for cytokeratin 19, was measured in 312 serum samples, including 164 lung cancer, 118 benign pulmonary disease, and 30 healthy individuals. The sensitivity of CYFRA 21-1 was also compared with two other markers, carcinoembryonic antigen (CEA) and squamous cell carcinoma antigen (SCC), in 164 patients with lung cancer.. The median value of healthy individuals was 1.3 ng/mL (95th percentile 1.8). In patients with benign pulmonary diseases, the median was 1.5 ng/mL (95th percentile 2.9). There is no significant difference between sexes, smoking habit, and the subgroups of benign pulmonary disease, such as tuberculosis, pneumonia, or COPD. Using the cutoff value of 3.3 ng/mL, defined at 95% specificity for benign lung disease, the sensitivities of CYFRA 21-1 for squamous cell carcinoma (n=74), adenocarcinoma (n=54), undifferentiated large cell carcinoma (n=11), and small cell lung cancer (n=25) were 62%, 39%, 36%, and 20%, respectively. Despite the cell types, the sensitivities of CYFRA 21-1 in non-small cell lung cancer (NSCLC, n=169) were 51% (CEA 42%, SCC 20%). The sensitivity of CEA was significantly higher in patients with adenocarcinoma (58%) than other markers; while in patients with squamous cell carcinoma, CYFRA 21-1 assay has the highest sensitivity. The median level of CYFRA 21-1 in squamous cell carcinoma is significantly higher than that of other cell types (Mann-Whitney test, p<0.001). The serum level and sensitivity of CYFRA 21-1 were well correlated with staging and tumor size in squamous cell carcinoma. The CYFRA 21-1 values were measured for monitoring progression of disease in 20 patients with squamous cell carcinoma. There is significant difference in paired observation of CYFRA 21-1 level in patients with progressive disease (Wilcoxon signed-rank test, p<0.05), but no difference was observed in patients with stabilized disease (p>0.1).. For patients with NSCLC, especially in squamous cell carcinoma, CYFRA 21-1 is not only a sensitive and specific tumor marker, but also may be a useful adjunctive marker for disease monitoring.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Disease Progression; Enzyme-Linked Immunosorbent Assay; Epithelium; Female; Humans; Keratins; Lung Diseases; Lung Diseases, Obstructive; Lung Neoplasms; Male; Neoplasm Staging; Pneumonia; Sensitivity and Specificity; Serine Proteinase Inhibitors; Serpins; Sex Factors; Smoking; Tuberculosis, Pulmonary

Bronchial epithelial dysplasia is a non-invasive cellular change often associated with physical or chemical injury and considered a pre-neoplastic lesion in the formation of lung cancer. A series of 39 bronchial dysplasias associated with both neoplastic and non-neoplastic lesions were assessed for expression of markers of differentiation by immunocytochemistry and compared with samples of normal bronchial epithelium. The normal bronchial epithelium studied expressed cytokeratins (CKs) 4, 6, 7, 8, 18, and 19 in all cases; CK 13 in 13 cases; and peanut agglutinin (PNA) in seven cases. Involucrin, CK 10, and CK 14 were not observed in the normal bronchial samples. In the dysplastic bronchial biopsies, epithelial staining was observed with epithelial CKs 7, 8, 18, and 19 in all cases; CK 13 was seen in 26 cases; CK 14 in 13 cases; CK 6 in 11 cases; and CK 10 in five cases. In 13 cases of dysplasia, only simple epithelial antigens were identified. Involucrin expression was observed in 17 dysplastic biopsies and PNA in 12. By Fisher's exact test, a significant association between non-severe histological grade of dysplasia and CK 6 expression (P = 0.018) was found. Comparison of the results using the same analysis showed significant correlations between the loss of CK 6 expression (P < 0.001) and the expression of CK 14 (P = 0.008) and involucrin (P = 0.0018) with bronchial dysplasia. These data show that the pattern of differentiation antigen expression in bronchial dysplasia is significantly different from that of the normal bronchial epithelium, but the phenotypic heterogeneity of these lesions is similar to that of bronchial carcinomas.

Topics: Aged; Aged, 80 and over; Antigens, Differentiation; Biomarkers; Bronchi; Cell Differentiation; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Lectins; Lung Neoplasms; Middle Aged; Peanut Agglutinin; Precancerous Conditions; Protein Precursors

To evaluate the diagnostic value of the serum cytokeratin 19 fragment (CYFRA 21-1) in bronchogenic carcinoma, we investigated the sera of 138 patients (58 with benign pulmonary disease and 80 with non-small cell lung cancer (NSCLC)) using immunoradiometric assay. The mean (SD) value of serum CYFRA 21-1 in NSCLC (13.26 (16.54)) was significantly higher than in benign lung diseases (1.74 (1.55)) (p < 0.0001). Sensitivity for CYFRA 21-1 (using 3.5 ng/ml, a cut-off value corresponding to a 95% specificity for benign pulmonary disease) in NSCL was 62%. Positive CYFRA 21-1 levels were significantly higher in 75% of patients with squamous cell carcinoma (n = 36) than in 53% with other NSCLC (n = 44) (p < 0.05). CYFRA 21-1 levels were significantly different between squamous cell carcinoma (17.28 (19.94)) and the other NSCLC (9.96 (12.44)) (P < 0.05). Elevated CYFRA 21-1 levels in patients with stage III and IV disease (n = 64, 18.19 (26.51)) were significantly higher than in stage I and II (n = 16, 4.41 (5.76)) (p < 0.02). The positive rate of CYFRA 21-1 in tumor stage I and II was only 37%. Our results indicate that CYFRA 21-1 may be a useful tumor marker in NSCLC, especially in squamous cell carcinoma. However, CYFRA 21-1 cannot be used for the diagnosis of early stage disease of NSCLC. CYFRA 21-1 may also contribute to the monitoring of NSCLC.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Peptide Fragments

The sensitivity and specificity of Cyfra 21-1 as marker for lung cancer was evaluated in comparison with carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC) and neuron-specific enolase (NSE). Patients with histologically verified lung cancer and different groups without lung cancer were investigated. Sensitivity of Cyfra 21-1 (cut-off level 2.9 micrograms/l) was 40% for non-small cell lung cancer (NSCLC), 60% for rare histological types and 21% for small cell lung cancer (SCLC). In NSCLC sensitivity of Cyfra 21-1 was 35% for squamous cell carcinoma and 41% for adenomous carcinoma. The highest sensitivity for CEA was 45% in NSCLC, with 57% in the subtype of adenomous cell carcinoma; for SCC 30% was achieved in squamous cell carcinoma and for NSE 66% sensitivity was reached in SCLC. In our patients Cyfra 21-1 and CEA appeared equally useful for evaluating patients with NSCLC.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Diagnosis, Differential; Humans; Immunoradiometric Assay; Keratins; Lung Diseases; Lung Neoplasms; Middle Aged; Neoplasm Staging; Phosphopyruvate Hydratase; Sensitivity and Specificity; Serpins

We examined two recently described cytokeratin markers, CYFRA 21-1 (cytokeratin fragment recognized by KS 19-1 and BM 19-21 antibodies) and TPS (specific M3 epitope of the tissue polypeptide antigen), in 405 lung cancer patients (91 small-cell and 314 non-small-cell lung cancers) and 59 patients presenting with nonmalignant pulmonary disease. Sensitivity-specificity relationship, as analyzed by receiver operating characteristic curves, demonstrated a higher accuracy of CYFRA 21-1 in comparison with TPS in both small-cell and non-small-cell lung cancers. Thresholds of 3.6 ng/ml and 140 U/L for CYFRA 21-1 and TPS respectively gave a 90% to 95% specificity. Sensitivity of CYFRA 21-1 was the highest in squamous-cell carcinomas (0.61) and the lowest in small-cell lung cancers (0.36), whereas sensitivity of TPS did not vary significantly according to histology (overall sensitivity, 0.40). In non-small-cell lung cancers, both serum CYFRA 21-1 and serum TPS distributions varied significantly according to Mountain's stage of the disease, nodal status, tumor status, and performance status, inasmuch as the worse each above-mentioned variable became, the higher the median and interquartile serum marker level was. Neither CYFRA 21-1 nor TPS was able to accurately discriminate between stage IIIa (marginally resectable) and stage IIIb (unresectable) non-small-cell lung cancers, however. In both small-cell and non-small-cell lung cancers, univariate survival analyses demonstrated that either a CYFRA 21-1 level over 3.6 ng/ml or a TPS level over 140 U/L significantly indicated a poor survival rate. In the whole population, taking into account other significant variables, Cox's model analysis demonstrated that a poor performance index, an advanced stage of the disease, the presence of metastases, elevated serum lactate dehydrogenase, and high serum CYFRA 21-1 (odds ratio, 1.74; 95% confidence interval, [1.33-2.27] were independent prognostic variables. We concluded that CYFRA 21-1 is a significant determinant of survival. Other applications of cytokeratin markers in lung cancer are still limited.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Female; Humans; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Peptides; Prognosis; Prospective Studies; Sensitivity and Specificity; Tissue Polypeptide Antigen

The CYFRA 21-1 assay is a test that has been developed recently for detection of a cytokeratin 19 fragment in serum. A diagnostic role for CYFRA 21-1 has already been proposed. The question of whether this marker is prognostically significant is important in clarifying the role of CYFRA 21-1 in clinical practice. The aim of this study was to evaluate the prognostic significance of elevated preoperative CYFRA 21-1 levels in patients with resected primary squamous-cell lung cancer (SqCC). Serum levels of CYFRA 21-1 were measured using an immunoradiometric assay (CIS bio) in 91 patients with operable SqCC. Survival and disease-free survival curves related to initial levels of this marker were estimated using the Kaplan-Meier method. In the univariate analysis the log-rank test and the log-rank test for trend were used. In the multivariate analysis the stratified log-rank test and the proportional hazard model were used. Elevated preoperative CYFRA 21-1 levels were identified in 55% of patients with SqCC. The number of patients with elevated levels of this marker increased with TNM stage (P = 0.0001). In univariate analysis elevated levels of CYFRA 21-1 were significantly associated with poor overall survival (P < 0.00005) and with disease-free survival (P < 0.00005). In multivariate analysis elevated levels of this marker were also found to be associated with poor overall and disease-free survival (P = 0.01 and P = 0.003 respectively). In conclusion, CYFRA 21-1 may be an independent prognostic parameter of survival and tumour relapse in SqCC and may be useful in identifying resected SqCC patients at high risk of treatment failure.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease-Free Survival; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged

Topics: Biopsy, Needle; Carcinoma; Fatal Outcome; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Middle Aged; Thyroglobulin; Thyroid Neoplasms

The aim of the study was to test the diagnostic value of serum tumor marker CYFRA 21-1 for squamous cell lung cancer (SQCLC) in comparison with carcinoembryonic antigen (CEA). Ninety-one patients were included in this study: 56 with SQCLC-Group I, 25 with other types of lung cancer-Group II, 10 with benign respiratory tract diseases-Group III. Median CYFRA 21-1 serum concentration (ng/ml) was: in Group I: 4.52 (0.94 - > 16), in Group II: 3.58 (1.72 - > 16), in Group III: 2.05 (0.99-3.41). Median CEA serum concentration (ng/ml) was: in Group I: 4.49 (0.76 - > 20), in Group II: 3.32 (1.17 - > 20), in Group III: 3.09 (1.84-6.37). There was a highly significant difference between the levels of CYFRA 21-1 in Group I and III (p < 0.001), but there was no statistically significant difference between the levels of CEA in Group I and III. Sensitivity of CYFRA 21-1 by the cut-off 3.33 ng/ml in the diagnostics of SQCLC was 0.68, specificity 0.90, positive predictive value 0.91, negative predictive value 0.65. Sensitivity of CEA by cut-off 4.61 ng/ml was 0.5 by the same specificity 0.90. CYFRA 21-1 has high sensitivity, specificity and positive predictive value in the diagnostics of SQCLC. Sensitivity of CYFRA 21-1 is significantly higher than sensitivity of CEA in this setting.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Sensitivity and Specificity; Statistics, Nonparametric

In order to improve the diagnosis of lung carcinoma, in which a complicated histologic pattern is present, the immunohistochemistry of 119 adenocarcinomas, 65 squamous cell carcinomas, 12 small cell carcinomas, 18 large cell carcinomas, and 15 metastatic adenocarcinoma in the lung were evaluated using a monoclonal antibody, KM195, against lung carcinoma, and compared with the immunohistochemical results using anti-human cytokeratin (CAM 5.2) and other monoclonal antibodies. Formalin-fixed, paraffin-embedded tissues were stained using the labeled streptavidin-biotin method. Extracts from fresh tissue homogenate, after fractionation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were transferred by Western blotting and stained with KM195. The anti-lung adenocarcinoma, murine, monoclonal antibody KM195 (IgG), was positive in 107 of 119 adenocarcinomas (90%), in 15 of 18 large cell carcinoma (83%), in three of 65 squamous cell carcinomas (5%), 13 of 15 (87%) metastatic adenocarcinoma in the lung, and was negative in 12 small cell carcinomas (P < 0.001). KM195-bound protein of primary and metastatic adenocarcinoma in the lung cases concentrated at about 40 kDa. In contrast, CAM 5.2 was positive in 52 of 67 (78%) adenocarcinomas, 10 of 62 (16%) squamous cell carcinomas, and was negative in six small cell carcinomas. These results suggest that the immunohistochemistry for KM195 may be a more useful marker over CAM 5.2 for the diagnosis of pulmonary adenocarcinoma.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Bacterial Proteins; Biomarkers, Tumor; Biotin; Blotting, Western; Carcinoembryonic Antigen; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Oligosaccharides; Paraffin Embedding; Reference Values; Sialyl Lewis X Antigen; Tissue Fixation

To evaluate the usefulness of CYFRA 21-1 and SCC Ag in the diagnosis of squamous cell carcinoma (SQC) of the lung, we tested sera from 124 patients with lung cancers (squamous cell ca 72, adenoca 22, large cell ca 4, small cell ca 18 and undetermined 8) and 78 patients with inflammatory lung diseases (bronchitis 24, bronchiectasis 29, tuberculosis 19 and others 6) using immunoradiometric assay kit for cytokeratin fragment 19 (CYFRA 21-1) and radioimmunoassay kit for SCC Ag. The serum CYFRA 21-1 and SCC Ag were significantly higher in lung cancer patients compared with control subjects. However, the significant difference was restricted only to SQC. In patients with SQC, CYFRA 21-1 and SCC Ag showed significantly higher levels according to the advanced anatomic stages (stage I-IIIa vs. stage IIIb, IV, p < 0.05). There was a good correlation between CYFRA 21-1 and SCC Ag (r = 0.41, p < 0.001). Receiver operating characteristic (ROC) curves were generated from results of both tumor markers and areas under the curves (AUC) were calculated. AUC of CYFRA 21-1 (0.93) were significantly larger than that of SCC Ag (0.77) for the diagnosis of SQC (p < 0.05). Therefore, we conclude that CYFRA 21-1 is superior to SCC Ag in the diagnosis of squamous cell carcinoma of the lung.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Humans; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Sensitivity and Specificity

Serum soluble cytokeratin 19 fragment (CYFRA) levels were measured in 251 patients with lung cancer and 139 patients with benign lung diseases to determine the clinical usefulness of CYFRA level determination in the diagnosis and monitoring of lung cancer. Serum levels of CYFRA were measured by a 2-step sandwich ELISA method. When the cut-off value was defined as 3.5 ng/ml, which was associated with a specificity of 95% for benign lung diseases, CYFRA had a high sensitivity (53%) in all patients with lung cancer. Both the serum level of CYFRA and its sensitivity increased significantly with the increase in clinical stage. A comparison of areas under receiver operating characteristic curves showed that CYFRA had the most power of discrimination in the diagnosis of lung cancer among markers including carcinoembryonic antigen, squamous cell carcinoma antigen, carbohydrate antigen 19-9, and neuron-specific enolase. A good correlation was found between serial changes in serum CYFRA levels during therapy and clinical responses for 18 patients who underwent chemotherapy and/or radiotherapy. Our findings suggest that CYFRA may be a marker of choice for screening and monitoring of lung cancer, particularly squamous cell carcinoma.

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; Carcinoma; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Lung Neoplasms; Male; Neoplasm Staging; Prognosis; Retrospective Studies; Sensitivity and Specificity; Survival Rate

The female genital tract is an infrequent site of metastasis, particularly for extragenital primaries. The ovary and vagina are the sites within the female genital tract that are the most frequently affected. The uterine corpus, especially the endometrium, is a distinctly unusual site of involvement. Primary lung cancer is the source of metastatic tumor to the female genital tract in less than 5% of patients. In the reported instances of endometrial involvement by a primary lung cancer, adenocarcinoma has been the reported subtype. Here, we report a case of well-differentiated neuroendocrine carcinoma of the lung metastatic to the endometrium in a 68-year-old woman with postmenopausal bleeding. Immunohistochemical studies were performed to confirm the neuroendocrine nature of the neoplasm.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Neuroendocrine; Endometrial Neoplasms; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Nerve Tissue Proteins

Mutations in the p53 gene are detected in greater than 50% of squamous cell carcinomas of the lung and to a lesser extent in adenocarcinomas. The p53 protein is also overexpressed in a relatively high percentage of preinvasive lesions of the bronchial epithelium. However, unlike tumor tissue, immunoreactivity does not necessarily imply that cells in preinvasive lesions carry a mutant p53 allele. In some cases, overexpression may result from a cellular checkpoint reaction to a toxic or mutagenic substance such as exposure to tobacco smoke. In any case, p53 overexpression in preinvasive lesions may serve as a biomarker for high risk assessment of lung cancer and other tumors in the aerodigestive tract. A study was designed to retrospectively analyze p53 overexpression in cells from sputum samples collected prior to histological tumor diagnosis. The rationale was based on the observation that both preinvasive and tumor cells from the bronchial epithelium are exfoliated into the airways and can be detected based on morphology in sputa. Two sets of cases were chosen: 1) patients whose first primary tumor was a squamous cell carcinoma containing a mutant p53 allele with overexpression observed in most of the tumor cells; and 2) patients whose squamous cell tumor did not contain a mutant p53 allele. Cells which stained positive for p53 expression were observed in sputum samples collected from all six patients whose tumors were positive for a mutant p53 allele. Also p53 positive cells were detected on sputum slides for two of the five cases where the tumor DNA did not contain a mutation and/or tumor cells which overexpress p53 were not detected in tissue sections. Although cells which stained positive for p53 were present in sputum from patients whose tumors contained a missense mutation, the presence of p53 overexpression was not specific for tumors which contain an altered p53 allele since overexpression was detected in sputum cells from patients whose tumor DNA did not contain a p53 mutation and/or tumor cells which stained positive for p53 were not observed in tissue sections. However, the p53 positive cells in sputa collected from the latter group of patients could have been exfoliated from other lesions which contained a mutant p53 allele. The accumulation of p53 in some sputum cells was concomitant with expression of simple epithelial type cytokeratins (CK) 8 and 18 or at least one of the other cytokeratins detected by a broad spectrum (PAN) CK antibody mi

Topics: Alleles; Carcinoma, Squamous Cell; Humans; Keratins; Lung Neoplasms; Mutagenesis; Retrospective Studies; Sputum; Tumor Suppressor Protein p53

This study was designed to find the differentiated phenotype and its clinicopathological significance in pulmonary large cell carcinoam (PLCC) by immunohistochemical methods using a panel of antibodies related to the phenotype differentiation: high molecular weight cytokeratin, low molecular weight cytokeratiin, secretory component, chromogranin A and synaptophysin in 60 cases of PLCC. The results demonstrated that all cases of PLCC possessed monophasic, biphasic and triphasic phenotype differentiation features respectively, including squamous (4 cases), adenomatous (20 cases), neuroendocrine (19 cases), adenosquamous (11 cases); and the the coexistence of adenomatous, squamous and neuroendocrine (6 cases). The median survival time was different between patients with various differentiated phenotypes of PLCC. There was a statistically significant difference (P < 0.05) in survival between neuroendocrine and adenomatous phenotypes. The results of this study implies that: (1) PLCC expressed different phenotypes of differentiation, (2) phenotypes of neuroendocrine differentiation has a poor prognosis, (3) it is necessary to classify PLCC according to the phenotype differentiation.

Topics: Carcinoma, Non-Small-Cell Lung; Chromogranins; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Phenotype; Survival Rate; Synaptophysin

Soluble cytokeratin fragment 19 levels were measured with an enzyme immunoassay method developed by Boehringer Mannheim (Enzymun-Test CYFRA 21-1) in the serum of 185 patients with lung cancer [149 with non-small-cell lung cancer (NSCLC) and 36 with small-cell lung cancer (SCLC)] and 97 patients with benign lung diseases in order to determine its clinical usefulness in the diagnosis of lung cancer and follow-up of treatment. We used the cut-off value of 3.5 ng ml-1, established by the Japan CYFRA research group. This cut-off value is based on calculations using the receiver operating characteristic approach instead of using the 95% specificity approach recommended by other authors. The resulting sensitivity and specificity for the group of all lung cancer patients were 65.4% and 84.5% respectively. The sensitivity was highest (76.1%) for squamous cell carcinoma and lowest (44.4%) for SCLC. For NSCLC patients, when CYFRA 21-1 levels were analysed by node (N) factor, patients who presented with mediastinal lymph node metastasis (N2 or N3) demonstrated higher serum CYFRA 21-1 levels (5.6; interquartile range 3.2-11.5 ng ml-1) than patients without mediastinal node metastasis (N0 or N1, 3.9; interquartile range 2.2-10.0 ng ml-1; Mann-Whitney U-test, P = 0.0373). We compared the discriminatory power of CYFRA 21-1 with that of other tumour markers including carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC) and neuron-specific enolase (NSE). The area under the curve (AUC) of each ROC curve was calculated using the CLABROC program for statistical analysis. CYFRA 21-1 appeared to have the most discriminatory power of the markers tested in the diagnosis of lung cancer. In serial measurements of 14 patients receiving chemotherapy or radiotherapy, a high degree of correlation was noted between serum levels of CYFRA 21-1 and extent of clinical response (Wilcoxon, P = 0.0093).

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Peptide Fragments

Cyfra 21-1 is a novel marker for non small cell lung cancer. Up to now only few data about the value of Cyfra 21-1 in the diagnosis of malignant and non-malignant pulmonary diseases are available. Further the effect of long-time cigarette consumption on serum concentrations of Cyfra 21-1 has not been described. Aim of this study therefore was the determination of Cyfra 21-1 in different malignant and non-malignant pulmonary diseases and in healthy smokers and nonsmokers.. Sera of healthy individuals (control group, n = 121; 63 smokers and 58 nonsmokers), patients with chronic bronchitis (n = 50), lung fibrosis (n = 38), exogen allergic alveolitis (n = 32), lung tuberculosis (n = 45), sarcoidosis (n = 30), small cell lung cancer (n = 60), squamous cell carcinoma (n = 53), non-small cell carcinoma (n = 29) and adenocarcinoma (n = 52) were analyzed.. Within the control group no significant differences of the Cyfra 21-1 serum concentration were observed between smokers and nonsmokers. Serum concentrations of Cyfra 21-1 were similar in the control group, patients with non-malignant pulmonary diseases and patients with small cell lung cancer whereas serum concentrations were significantly increased in patients with non-small cell lung cancer, adenocarcinoma and squamous cell carcinoma.. The data confirm the high sensitivity and specific of Cyfra 21-1 for the differential diagnosis between malignant and non-malignant pulmonary diseases as well as small cell and non-small cell lung cancer.

Topics: Adolescent; Adult; Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Diagnosis, Differential; Female; Humans; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Reference Values; Smoking

Ultrastructural examinations have shown myofibroblastoid differentiation in sarcomatoid/desmoplastic mesotheliomas, but immunohistochemical expression of muscle actins seldom has been documented. We examined 10 sarcomatoid, 12 epithelial, and five biphasic mesotheliomas immunohistochemically for the expression of muscle-specific actin (MSA) and smooth muscle actin (SMA) and compared it with that in 12 specimens of lung cancer. All of the sarcomatoid mesotheliomas were found to be positive for both MSA and SMA. The epithelial cells in nine epithelial and two biphasic mesotheliomas were positive for MSA, but SMA was only positive in one epithelial mesothelioma. Conversely, the lung cancers were negative for both MSA and SMA in the epithelial cells, except for one specimen that was weakly positive for MSA. The stromal cells in both the epithelial mesotheliomas and lung cancers were negative for cytokeratin but were positive for MSA and SMA, whereas the sarcomatoid and biphasic mesothelioma spindle cells were positive for all three antibodies. We concluded that sarcomatoid mesothelioma was positive for MSA and SMA, which is in support of its myofibroblastic differentiation, and that positivity for MSA in some epithelial mesotheliomas might be of diagnostic value in differentiation from lung cancers.

Topics: Actins; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mesothelioma; Muscle, Smooth; Muscles; Staining and Labeling

We wanted to investigate the diagnostic and prognostic significance of serum CYFRA 21-1, especially in predicting the risk of recurrence in patients with operable squamous cell lung cancer. Serum levels of CYFRA 21-1 were measured using an immunoradiometric assay (CIS bio) in 76 patients with squamous cell lung cancer (64 operable and 12 with unresectable tumours), 22 with other non-small-cell type (12 with adenocarcinoma and 10 with large-cell type) and 45 with nonmalignant lung diseases. Elevated preoperative CYFRA 21-1 levels were identified in 63% of patients with squamous cell type (SqCC), 33% with adenocarcinoma, and 30% with large-cell carcinoma type. The diagnostic specificity of the assay was 96%. Positive CYFRA 21-1 levels were observed in 33% of stage I, 52% of stage II, 76% of stage IIIa and 83% of stage IIIb patients with SqCC type. Statistically significant differences were obtained between stages I and II and between II and IIIa, but not between stages IIIa and IIIb. Recurrence-free survival probability for patients with elevated serum CYFRA 21-1 levels before surgery was 63% (24/38) versus 92% (24/26) for patients with normal serum CYFRA 21-1 levels. However, the difference was not statistically significant when adjusted for the TNM stage (primary tumour, regional lymph node involvement, occurrence of distant metastasis). In 9 of the 10 patients with increased trend for CYFRA 21-1 during follow-up, elevated serum CYFRA 21-1 levels preceded (7) or coincided (2) with the clinical detection of tumour recurrence, providing a predictive value of an increased trend of 90%.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Humans; Immunoradiometric Assay; Keratins; Lung Neoplasms; Multivariate Analysis; Neoplasm Recurrence, Local; Prognosis; Proportional Hazards Models; Risk Factors; Sensitivity and Specificity; Survival Analysis

Although the general conception is that hepatocyte- and bile duct-specific cytokeratin (CK) patterns are maintained throughout the neoplastic process, there is an increasing number of reports showing deviation from the rule. CK patterns have been found to be similar across species barriers, so it could be expected that studying the CK patterns of experimentally induced liver tumors may contribute to the elucidation of these controversies.. A CK immunohistochemical study was carried out on histologic sections from hepatocellular carcinomas (HCCs) and preneoplastic lesions from 118 monkeys chronically dosed with diethylnitrosamine (DEN), using mAbs to CK 8, CK 18, CK 7, and CK 19.. Normal monkey hepatocytes differed from human hepatocytes by displaying CK 19 in addition to the CK 8/CK 18 pairs, whereas the CK pattern of the bile duct epithelial cells was identical in monkey and human liver. In association with DEN-induced hepatocarcinogenesis, heterogeneity was observed in the CK expression, both in the HCCs and nontumorous parts of the livers. The majority of the HCC cases displayed one of the three CKs normally present in monkey hepatocytes, whereas positive expression of all three CKs (CK 8, CK 18, CK 19) and negative CK 7 was preserved in only 19.5% of the HCC cases. A so-called mixed staining pattern (negative and positive CK staining within the same tumor) was observed in approximately one-fourth of the cases. There was no correlation between the preservation of the hepatocyte-specific CK pattern and the degree of differentiation, tumor grade, or DNA ploidy of the HCCs. In approximately 10% of the primary tumors, CK 7 was expressed in the entire parenchymal cell compartment of the HCC nodules, whereas it was present in a mixed staining pattern in more than half of the cases. In lung metastases, CK 7 was less common, only expressed in approximately one-fourth of the cases. Alterations in the CK patterns were observed in the nonneoplastic hepatocytes of the tumor-bearing monkeys. These included mixed staining patterns in which the CKs appeared as positive and negative regenerating nodules side-by-side. As was observed in the HCCs, CK 7 was more commonly expressed in the nonneoplastic parenchyma in the form of mixed staining pattern than the other three CKs. Moreover, CK 7-negative HCCs occurred more frequently in CK 7-negative livers than in positive livers. Proliferation of CK 7- and CK 19-positive bile ductules and bile ductular-like (oval) cells was frequently associated with the DEN-induced liver injury and hepatocarcinogenesis.. This is the first report on CK expression in monkey liver. The findings show that the hepatocyte specific pattern is not always preserved during DEN-induced hepatocarcinogenesis and may therefore not be useful in differentiating between HCCs and cholangiocarcinomas.

Topics: Animals; Antibodies, Monoclonal; Carcinoma, Hepatocellular; Chlorocebus aethiops; Diethylnitrosamine; Immunohistochemistry; Keratins; Liver Neoplasms; Lung Neoplasms; Macaca fascicularis; Macaca mulatta; Reference Values

Despite extensive research, the role of the commonly employed tumour markers in the diagnosis of lung carcinoma is yet to be clarified. The utility of a new marker, CYFRA 21-1, in the preoperative evaluation of patients with bronchogenic carcinoma was investigated. CYFRA 21-1 was determined with a radiometric assay in serum of 280 patients with lung cancer and 208 patients with various nonmalignant lung diseases. The levels of the marker were significantly higher in lung cancer patients. Among benign lung diseases, elevated CYFRA 21-1 levels were found in pulmonary fibrosis. Using a cut-off of 3.2 ng.ml-1 (95th percentile of levels obtained in benign lung disease), the total sensitivity of the marker was 48%. The best sensitivity was obtained in squamous cell lung cancer (60%). The highest values of CYFRA 21-1 were found in metastatic lung cancer, and the marker sensitivity was more elevated in stage IIIb and IV. On the other hand, 40% of patients with surgically resectable lung cancer had CYFRA 21-1 levels above the cut-off. We conclude that CYFRA 21-1 may be satisfactorily employed in the differential diagnosis between malignant and benign lung diseases in association with other clinical and radiological data.

Topics: Biomarkers, Tumor; Carcinoma, Bronchogenic; Diagnosis, Differential; Female; Humans; Immunoradiometric Assay; Keratins; Lung Diseases; Lung Neoplasms; Male; Pulmonary Fibrosis; Sensitivity and Specificity

In this study we compared the diagnostic utility of: (1) neuron-specific enolase (NSE); (2) squamous cell carcinoma antigen (SCC); (3) carcinoembryonic antigen (CEA); and (4) cytokeratin markers (CYFRA 21-1, TPA, TPM, TPS) in patients with small-cell lung cancer (SCLC) (21 cases) and non-small-cell lung cancer (94 cases). For comparison we also studied 66 patients with benign lung diseases and nine with pleural mesothelioma. NSE levels in SCLC patients were significantly higher than those in all the other groups studied. No significant variations were found among the SCC levels in all groups. CEA levels in patients with adenocarcinoma were significantly higher than those in all other groups studied. CYFRA 21-1 serum levels significantly increased in patients with squamous cell carcinoma and mesothelioma, while TPA, TPS and TPM increased in patients with lung cancer irrespective of the histological type. In patients with SCLC, high levels of all markers except SCC were found when the disease was extensive. In patients with non-SCLC, the highest levels of all tumour markers were usually found in those with advanced disease, although CYFRA 21-1 gave a sensitivity of 44% when a specificity of 95% was fixed in stage I non-SCLC patients. An analysis of receiver operating characteristic curves revealed that the highest diagnostic accuracies in distinguishing benign from malignant lung diseases were achieved with TPM (81%), CYFRA 21-1 (72%), CEA (78%) or TPA (78%) when using cut-off values of 46 Ul-1, 3.0 micrograms l-1, 2.0 micrograms l-1 and 75 Ul-1 respectively. When all patients were considered, the combined evaluation of more than one marker did not significantly improve the results obtained with TPM alone. However, taking into consideration the fact that CYFRA 21-1 is the most sensitive index of early lung tumours and that its combined determination with TPM did not worsen the overall sensitivity and specificity of the latter, the combined use of these two markers may be suggested as a useful took for the diagnosis of lung tumours.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Peptides; Phosphopyruvate Hydratase; Serpins; Tissue Polypeptide Antigen

Cytokeratins are epithelial markers whose expression is not lost during malignant transformation. Cyfra 21-1 is a cytokeratin-19 fragment that is soluble in serum and may be a useful circulating tumor marker.. The aims of this study were (1) to confirm sensitivity and specificity of Cyfra 21-1 in detecting non-small cell lung cancer (NSCLC) and especially the squamous cell subtype, (2) to assess the potential relationship between Cyfra 21-1 and disease stage of the disease in NSCLC, and (3) to evaluate prognostic effect of Cyfra 21-1 in NSCLC.. An immunoradiometric assay of serum Cyfra 21-1 was performed in 161 patients with lung cancers and 71 others with benign lung diseases. The ability of Cyfra 21-1 to detect different histologic subtypes of lung cancer vs benign lung diseases was assessed through receiver operating characteristic (ROC) curves and comparisons with other tumor markers such as carcinoembryonic antigen, neuron-specific enolase, and squamous cell carcinoma antigen. Comparisons of Cyfra 21-1 levels according to histologic subtype and disease stage were done using Kruskal-Wallis test. Independent prognostic value of Cyfra 21-1 was studied with a multivariate analysis of survival (Cox's model).. Using a threshold of 3.3 ng/mL for Cyfra 21-1, sensitivity and specificity were, respectively, 0.59 and 0.94 in NSCLC, 0.68 and 0.94 in the subgroup of the squamous cell carcinoma, and 0.19 and 0.94 in small cell lung cancer. Cyfra 21-1 levels were significantly higher in advanced NSCLC than in early-stage disease. All 29 patients with serum concentrations > 32 ng/mL had stage IIIB-IV and only one of 14 patients with stage I-II disease had Cyfra 21-1 level > 18 ng/mL. In the multivariate analysis of survival, Cyfra 21-1 was an independent prognostic factor along with performance status and disease stage in NSCLC.. Cyfra 21-1 is a sensitive and specific tumor marker of NSCLC, especially of squamous cell subtype. It also reflects the extent of the disease and has an independent prognostic role along with performance status and disease stage in NSCLC.. A high level of Cyfra 21-1 in apparently early-stage NSCLC should be an indication for more extensive workup before thoracotomy. The independent prognostic role of Cyfra 21-1 level may be useful in stratifying populations with advanced NSCLC or early-stage resected NSCLC as elevated Cyfra 21-1 levels might identify those patients at high risk for treatment failure.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Immunoradiometric Assay; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Prospective Studies; ROC Curve; Sensitivity and Specificity

Pulmonary blastoma is a rare malignant neoplasm that histologically resembles airway structures and mesenchymal supporting tissues seen in the early phases of fetal lung development. The tumor either is biphasic with immature epithelial and stromal components or consists solely of an epithelial component. The preferred terminology for the latter is pulmonary endodermal tumor resembling fetal lung. Pulmonary blastoma is usually peripheral in location, and rarely does this tumor present intrabronchially. We report a case of pulmonary endodermal tumor resembling fetal lung presenting as a polypoid intrabronchial mass.

Topics: Adenocarcinoma; Adult; Animals; Carcinoembryonic Antigen; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Membrane Glycoproteins; Mucin-1; Mucins; Pulmonary Blastoma

We reviewed 45 pulmonary B-cell non-Hodgkin's lymphomas to determine whether their morphology and immunohistochemical features were those of lymphomas arising from mucosa-associated lymphoid tissue (MALT), as described in other sites. The polymerase chain reaction was used to provide further information on clonality. We found that these lymphomas infiltrate the pulmonary interstitium along bronchovascular bundles and interlobular septa, subsequently spilling out into airspaces and finally destroying the alveolar architecture of the lung. Central hyaline sclerosis and vascular infiltration were common features. All lymphomas stained CD20 positive and were accompanied by variable numbers of reactive CD3 positive T-cells. Cytokeratin staining with CAM 5.2 was useful in identifying lymphoepithelial lesions. CD21 staining of follicular dendritic cells revealed germinal centres where they were not recognizable on H & E staining. The polymerase chain reaction was performed on paraffin tissue from 28 patients. Twenty were low grade, of which 12 showed a clonal band and eight showed a polyclonal smear pattern. Eight were high grade, of which one revealed a clonal band. Three produced polyclonal smear patterns and four cases were inadequate samples. In one patient who had lymphoma at a second extranodal site, identical bands were identified, evidence for 'homing' of lymphoid cells towards mucosal epithelium.

Topics: Base Sequence; CD3 Complex; DNA, Neoplasm; Humans; Immunoglobulin kappa-Chains; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Lymphoma, B-Cell; Lymphoma, B-Cell, Marginal Zone; Molecular Sequence Data; Neoplasms, Second Primary; Polymerase Chain Reaction; Pulmonary Alveoli; Receptors, Complement 3d; Sclerosis; T-Lymphocytes

A large proportion of patients with operable lung carcinoma (no evidence of systemic spread of tumor) develop metastatic disease after primary therapy. More sensitive and specific methods are needed to identify patients at highest risk for recurrence who may benefit most from adjuvant therapy, while sparing those patients who do not require such treatment.. Using epithelial-specific monoclonal antibodies, the authors have developed an immunocytochemical assay capable of detecting as few as 2 lung cancer cells in 1 million bone marrow cells.. The assay was used to test the bone marrow (from resected ribs) of 43 patients with primary non-small cell lung carcinoma who showed no clinical or pathologic evidence of systemic disease.. Occult bone marrow micrometastases (BMMs) were detected in 40% of patients (17/43) with non-small cell lung cancer, including 29% (5/17) of patients with stage I or II disease and 46% of whom (12/26) had stage III disease. The median follow-up was 13.6 months. Patients with occult BMMs had significantly shorter times to disease recurrence compared with patients without BMMs (7.3 vs. > 35.1 months, p = 0.0009). Furthermore, for patients with stage I or II disease, the presence of occult BMMs was significantly associated with a higher rate of recurrence (p = 0.0004).. The detection of occult BMMs identifies patients with operable non-small cell lung carcinoma who are at significantly increased risk for recurrence, independent of tumor stage, and may be useful in evaluating patients for adjuvant treatment protocols.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Bone Marrow Neoplasms; Carcinoma, Non-Small-Cell Lung; Epithelium; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Neoplasm Recurrence, Local; Risk Factors; Survival Rate

A correct diagnosis of small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) is essential both for prognostic and therapeutic reasons. We used discriminant analysis as a method to optimize the discriminant power of serum tumour marker levels for differentiation between SCLC and NSCLC. A panel of serum markers, including neurone specific enolase (NSE), cytokeratin fragment antigen 21.1 (CYFRA-21.1), tissue polypeptide antigen (TPA) and carcinoembryonic antigen (CEA) was obtained in 50 consecutive NSCLC and 17 SCLC. Data were analysed by the BMDP statistical program after logarithmic transformation of marker levels. The variables selected were NSE and CYFRA-21.1. Considered together, they were able to give a 97% rate of correct classification. The formula generated (canonic variable, CV) was validated on a group of seven SCLC and 22 NSCLC patients. Only two errors occurred. We therefore conclude that the canonic variable tested, based on NSE and CYFRA-21.1, provides a good discrimination between the two types of lung cancer. The method is rapid, relatively inexpensive, and based on simple serum tests.

Topics: Aged; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Diagnosis, Differential; Discriminant Analysis; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Peptides; Phosphopyruvate Hydratase; Tissue Polypeptide Antigen

Sensitive detection of systemic tumor dissemination in lung cancer patients is important for selection of appropriate treatment modalities. Based on recent promising data that showed reverse transcriptase-polymerase chain reaction (RT-PCR) analyses for cytokeratin 19 (CK-19) expression in peripheral-blood or bone marrow samples to be a rapid and highly sensitive method for detection of hematogenous tumor dissemination in patients with breast and prostate cancer, we evaluated the specificity of this assay system in lung cancer patients and a large number of healthy controls.. We examined CK-19 mRNA expression by RT-PCR in 17 lung cancer cell lines and in peripheral-blood samples of 50 lung tumor patients and 65 healthy controls.. Expression of CK-19 mRNA was observed in all lung cancer cell lines and in 50% of peripheral-blood samples from lung tumor patients. However, under the experimental conditions analyzed, at least 20% of the control samples were positive for CK-19 mRNA expression.. Contrary to prior reports, RT-PCR may detect non-tissue-specific constitutive low-level (illegitimate) expression of CK-19 mRNA in peripheral-blood mononuclear (PBMN) cells in a significant number of healthy controls. This finding may not only hamper the use of this assay system in lung cancer patients, but also questions its proposed applicability in patients with other epithelial tumors such as breast and prostate cancer.

Topics: Adult; Aged; Base Sequence; Female; Humans; Keratins; Leukocytes, Mononuclear; Lung Neoplasms; Male; Middle Aged; Molecular Sequence Data; Neoplasm Metastasis; Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Transcription, Genetic

The aim of this study was to evaluate serial determinations of CYFRA 21-1 in the follow-up of patients treated surgically for non-small cell lung cancer in order to predict the risk of tumour recurrence. Serum levels of CYFRA 21-1 were measured using an immunoradiometric assay (CIS bio) in 57 patients with operable non-small cell lung cancer (NSCLC): 25 with squamous cell carcinoma (SqCC), 20 with adenocarcinoma (AC), 12 with large cell carcinoma (LCC) and 30 with non-malignant lung diseases. Elevated preoperative CYFRA 21-1 levels were identified in 44% of all patients with NSCLC. The diagnostic specificity of the assay was 97%. Positive CYFRA 21-1 levels was observed in 30% of stage I, 33% of stage II, and 55% of stage IIIa. Statistically significant differences were obtained between stages I and IIIa, II and IIIa, but not between stages I and II. During follow-up recurrence was observed in 19 of 57 (33%) NSCLC patients. Recurrence-free survival probability for patients with elevated serum CYFRA 21-1 levels before surgery was 52% (13/25), versus 81% (26/32) for patients with normal serum CYFRA 21-1 levels (p < 0.01). In 15 patients with increased trend for CYFRA 21-1, elevated serum CYFRA 21-1 levels preceded (13 patients) or coincided (2 patients) with the clinical detection of tumour recurrence, providing a predictive value of an increased trend of 87%. In the multivariate analysis the association of the increase of CYFRA 21-1 level with a higher risk of recurrence is statistically significant (p < 0.001).

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Humans; Immunoradiometric Assay; Keratins; Lung Neoplasms; Multivariate Analysis; Neoplasm Recurrence, Local; Predictive Value of Tests; Prognosis; Sensitivity and Specificity; Survival Rate

There are few specific pathologic findings that can be relied on to distinguish primary thymic carcinomas from lung carcinomas with mediastinal extension or showing metastasis to the anterior mediastinum. The immunohistochemical reactivity on frozen sections of thymic carcinomas and lung carcinomas, which are histologically similar to each other, was examined with the use of monoclonal antibodies to cytokeratins 7 and 13. Among keratinizing squamous cell carcinomas, all thymic carcinomas reacted with antibody specific for cytokeratin 7 (9/9, 0%), whereas no staining reaction was seen in lung carcinomas (0/5, 0%) (p < 0.01). This finding can be used as a diagnostic aid in primary thymic keratinizing squamous cell carcinomas to expedite treatment and prognosis. Cytokeratin 7 and cytokeratin 13 monoclonal antibodies reacted with almost all cases of thymic carcinoma. Applications of monoclonal antibodies specific for certain cytokeratins, especially 7 and 13, may be helpful in the diagnosis of other subtypes of thymic carcinomas.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Thymoma; Thymus Gland; Thymus Neoplasms

Topics: Carcinoembryonic Antigen; Carcinoma; Carcinoma, Squamous Cell; Chemotherapy, Adjuvant; Follow-Up Studies; Humans; Hypopharyngeal Neoplasms; Keratins; Laryngeal Neoplasms; Liver Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasms, Multiple Primary; Neoplasms, Second Primary; Paget Disease, Extramammary; Penile Neoplasms; Skin Neoplasms

A case of metastatic bronchogenic carcinoma to the gingiva in a 47-year-old male is reported. The gingival lesion developed as a quickly growing mass and appeared 2 months after surgical excision and radiotherapy of the lung carcinoma were completed. The gingival tumor was histopathologically diagnosed as a poorly differentiated squamous carcinoma. Comparative cytologic studies showed similarities between the gingival metastasis and the previous lung cancer.

Topics: Antigens, Neoplasm; Carcinoma, Bronchogenic; Carcinoma, Squamous Cell; Gingival Neoplasms; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Mitosis; Mucin-1; Neoplasm Proteins

Evidence exists that apoptosis or programmed cell death plays an important role in tumor growth. Morphologically apoptosis is characterized by membrane blebbing and nuclear fragmentation in individual cells. In this study, we investigated changes in the nuclear organization in spontaneously occurring apoptotic cells in several cancer cell lines using several antibodies to nuclear matrix constituents. It appeared that nuclear matrix components remained detectable in cells undergoing spontaneous apoptosis. The same results were found when apoptosis was induced by cycloheximide in the non-small cell lung cancer cell line MR65. Using this induction method, the percentage of apoptotic cells in MR65 cells increased, allowing a more detailed and extensive examination of nuclear matrix alterations together with cytoskeletal changes. To study the expression of cytokeratins, type A- and B lamins, a nuclear matrix-associated 13 kDa U1RNP particle and the Ki67-antigen, immunocytochemistry in combination with confocal scanning laser microscopy was used. Apoptotic cells were identified based on nuclear morphology and the in situ nick translation assay. Whereas immunoreactivity against lamins and Ki67-Ag was rapidly lost during apoptosis, expression of the 13 kDa protein and, in early apoptotic stages, also cytokeratin expression, was observed to remain present. Dead cells lacked reactivity with all the antibodies tested. The persistence of nuclear matrix components is therefore a useful marker for the detection of apoptosis.

Topics: Antigens, Nuclear; Apoptosis; Cycloheximide; Cytoskeletal Proteins; DNA; Genetic Techniques; Humans; Keratins; Lung Neoplasms; Nuclear Proteins; Staining and Labeling; Tumor Cells, Cultured

This report represents an attempt to combine the serum levels of more tumor markers together to evaluate the response to chemotherapy in 26 patients affected with small cell lung cancer (SCLC), by means of discriminant analysis. A pilot prospective study was performed on 26 subjects affected with inoperable SCLC (18 extensive diseases, and 8 limited diseases) and treated with chemotherapy (etoposide plus cisplatin regimen). Serum levels of a panel of tumor markers: Carcinoembryonic antigen (CEA), Tissue Polypeptide Antigen (TPA), Neuron Specific Enolase (NSE) and CYFRA -21.1 were determined before starting chemotherapy and at the restaging time (after 3 months). To optimize the classification power of these markers, a discriminant analysis was done, which permitted generating two classification functions, based on Tissue Polypeptide Antigen and Neuron Specific Enolase levels able to correctly classify 25 out of 26 subjects (8 progressions and 18 non progressions). The results obtained, furtherly confirm that tumor markers are useful to evaluate the chemotherapy response and indicate a possible approach to obtain the maximum usefulness of the serum marker levels.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Small Cell; Discriminant Analysis; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Peptides; Phosphopyruvate Hydratase; Pilot Projects; Prospective Studies; Tissue Polypeptide Antigen

CEA, SCC and CYFRA 21-1 were measured in samples of serum coming from 105 'Non small cell lung cancer' (NSCLC) patients. The present study has been carried out to compare these markers, to analyse their prognostic significance and to determine the best combination of tumor markers. The median value and interquartile range were: CYFRA 21-1: 2,3 ng/ml, CEA: 3,7 ng/ml, SCC: 1,2 ng/ml. CEA demonstrated higher values in adenocarcinomas (P = 0.04). SCC and CYFRA 21-1 were comparable in the different histologic groups. CYFRA 21-1 and CEA values were dependant on tumor stage. Advanced tumors (T3 and T4) demonstrated higher serum CYFRA 21-1 level (P = 0.0006). CYFRA 21-1 was higher than 3,3 ng/ml in 36% of patients. CEA was higher than 5 ng/ml in 38% of patients and SCC was higher than 2 ng/ml in 27% of patients. Patients with a high CEA and CYFRA21-1 serum level had a shorter survival than those with a normal serum level. In a Cox regression analysis four variables (TNM stage, age, CYFRA 21-1 and CEA level) were found to be significant in the prediction of survival; CYFRA 21-1 level had the lowest P value (P = 0.0002). The current study suggests the use of a combination of CEA and CYFRA 21-1 in the clinical care of NSCLC.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Neoplasm Staging; Predictive Value of Tests; Prognosis; Proportional Hazards Models; Serpins; Survival Analysis

The analysis of the cell cycle distributions by univariate flow cytometric DNA measurement has been widely applied in the clinic to determine kinetic parameters of human malignancies. A common problem with measurements of cell cycle phase distributions in tumor biopsy material is the presence of non-malignant diploid cells. Furthermore, such a static measurement might not be accurate enough to describe the dynamic process of cell proliferation. For this purpose alternative methods have been developed to include BrdUrd incorporation or the presence of intrinsic proliferation associated markers such as PCNA or Ki67-Ag into the analysis. However, the presence of nonmalignant diploid cells will influence also these bivariate analyses, especially in case of DNA-diploidy of the tumor cells. Here we present a three parameter flow cytometric assay based on the simultaneous detection of cytokeratin, DNA and a proliferation associated marker, such as BrdUrd, PCNA or Ki67-Ag. Based on the presence of cytokeratin, epithelial cells can be selected for a detailed cell cycle analysis. This method can be applied to frozen tissue, which makes this assay useful for multicentre clinical studies.

Topics: Analysis of Variance; Antibodies; Biomarkers; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Cell Cycle; Cell Division; Cell Line; DNA, Neoplasm; Flow Cytometry; Humans; Interphase; Keratins; Ki-67 Antigen; Lung Neoplasms; Microscopy, Confocal; Mitosis; Neoplasm Proteins; Nuclear Proteins; Nucleic Acid Denaturation; Proliferating Cell Nuclear Antigen; Regression Analysis; Tumor Cells, Cultured

High resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) is a powerful research tool for the analytical separation of cellular proteins. The qualitative and quantitative pattern of polypeptides synthesized by a cell represents its phenotype and thus defines characteristics such as the morphology and the biological behavior of the cell. By analyzing and comparing the protein patterns of different cells it is possible to recognize the cell type and also to identify the most typical features of these cells. In applied pathology it is often difficult to identify the tissue of origin and the stage or grade of a neoplasia by cellular morphology analyzed by classical or immunostaining procedures. The protein pattern itself is the most characteristic feature of a cell and should therefore contribute to the identification of the cell type. For this reason we separated protein fractions originating from different lung tumor cell lines using 2-D PAGE and we compared the resulting patterns on a multivariate statistical level using correspondence analysis (CA) and ascendant hierarchical clustering (AHC). The results indicate that (i) protein patterns are highly typical for cells and that (ii) the comparison of the protein patterns of a set of interesting cell types allows the identification of potentially new marker proteins. 2-D PAGE is thus a unique and powerful tool for molecular cytology or histopathology, unveiling the protein expression level of tissues or cells.

Topics: Adenocarcinoma; Cytoskeletal Proteins; Electrophoresis, Gel, Two-Dimensional; Humans; Immunoblotting; Keratins; Lung Neoplasms; Membrane Proteins; Mesothelioma; Multivariate Analysis; Neoplasm Proteins; Tumor Cells, Cultured; Vimentin

Twenty-eight epithelial and 22 nonepithelial feline tumors were studied immunohistochemically. Epithelial tumors were 10 squamous cell carcinomas, two basal cell tumors, two sebaceous gland carcinomas, three apocrine gland carcinomas, three thyroid papillary carcinomas, one thyroid solid carcinoma, one renal clear cell carcinoma, one renal papillary carcinoma, one endometrial carcinoma, and four lung bronchioloalveolar carcinomas. Nonepithelial tumors were 10 fibrosarcomas, one liposarcoma, one leiomyosarcoma, one rhabdomyosarcoma, one hemangiosarcoma, two mast cell tumors, one osteosarcoma, three melanomas, and two lymphomas. Commercially available antibodies directed against high- and low-molecular-weight keratins (keratin, RCK-102, NCL-5D3), vimentin, desmin, glial fibrillary acidic protein (GFAP), and neurofilament intermediate filament (IF) proteins were used in the avidin-biotin-peroxidase complex technique on formalin-fixed, paraffin-embedded tumor tissue samples. All epithelial tumors except the endometrial carcinoma expressed some type of keratin protein. Squamous cell carcinomas expressed high-molecular-weight keratins exclusively. Coexpression of high- and low-molecular-weight keratins was observed in one basal cell tumor, sebaceous and apocrine adenocarcinomas, and thyroid, renal, and lung carcinomas. In addition to keratins, vimentin immunoreactivity was found in all basal cell tumors, all sebaceous gland, thyroid papillary, renal, and lung adenocarcinomas, and one of the apocrine gland adenocarcinomas. Immunoreactivity with GFAP antibody was found in one basal cell tumor and one sebaceous gland adenocarcinoma. The endometrial carcinoma did not react with any of the antibodies applied. Nonepithelial tumors analyzed expressed either vimentin (fibrosarcomas, liposarcoma, haemangiosarcoma, mast cell tumors, osteosarcomas, melanomas) or vimentin and desmin (leiomyosarcoma, rhabdomyosarcoma, one fibrosarcoma) IF proteins exclusively. Lymphomas did not react with any of the antibodies employed. These findings indicate that IF proteins antibodies can be included in diagnostic panels of antibodies for immunocharacterization of feline tumors. In addition, they can be used as a basis for the diagnoses of poorly differentiated or undifferentiated feline neoplasms.

Topics: Animals; Cat Diseases; Cats; Desmin; Glial Fibrillary Acidic Protein; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Kidney; Kidney Neoplasms; Lung Neoplasms; Neoplasms; Neoplasms, Glandular and Epithelial; Neurofilament Proteins; Skin Neoplasms; Thyroid Gland; Thyroid Neoplasms; Vimentin

Granular cell tumor (GCT) is a morphologic designation for tumors of varied histogenesis. Most GCTs in human beings are derived from Schwann cells, and rat meningeal GCTs are believed to originate in the neural crest. Three equine pulmonary GCTs from aged horses were studied immunohistochemically with primary antibodies directed against vimentin, cytokeratins (AE1/AE3), S-100, Leu 7, desmin, and neuron-specific enolase (NSE) using a steptavidin-biotin procedure. All three tumors stained similarly with strong and diffuse staining of neoplastic cells for vimentin and S-100 and negative staining with all other antibodies. On the basis of the immunohistochemical results and the previously described histologic and ultrastructural characteristics, equine pulmonary GCT is designated as neural crest and possibly Schwann cell derived, similar to GCT in rats and human beings.

Topics: Animals; Cell Transformation, Neoplastic; Desmin; Female; Granular Cell Tumor; Horse Diseases; Horses; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Phosphopyruvate Hydratase; Schwann Cells; Vimentin

The aim of our work was the evaluation of the immunoradiometric assay (IRMA) of two cytokeratinic markers, TPS and CYFRA 21.1, in clinical setting on non small cell lung cancer (NSCLC). Serum samples were obtained from 148 untreated NSCLC patients, 60 patients with non malignant lung diseases and 100 healthy subjects: TPS and CYFRA 21.1 serum levels were assayed by IRMA methods. Diagnostic performance of the markers was evaluated and the TPS and CYFRA 21.1 distribution analysed according to some different clinical and biological variables as histological subtypes, stage and survival time by using the Mann-Whitney "U"-test. Sensitivity, specificity and accuracy were 0.54 (80/148), 0.47 (28/60), 0.52 (108/208) and 0.73 (108/148), 0.74 (44/60), and 0.73 (152/208) for TPS and CYFRA 21.1 respectively. CYFRA 21.1 demonstrate a higher sensitivity than TPS in all stages of the disease and in the spinocellular and adenocarcinoma histological subtypes while TPS sensibility is higher in large cell carcinoma. The CYFRA 21.1 specificity is better than TPS probably by reason of its preferential distribution in respiratory epithelium. Both markers serum levels differ significantly between Stage I-II and IV and between Stage I-II-IIIa and IIIb-IV but neither TPS nor CYFRA 21.1 can discriminate Stage IIIa from IIIb. No significant differences were found in the serum expression of the markers by the different histological subtypes. A value of both markers less than the selected cut-off is related to a longer survival of the patients apart from therapy (p < 0.05). Our conclusion supports similar behaviour of these markers in NSCLC and indicates CYFRA 21.1 as the more needed biochemical index to evaluate NSCLC patients.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Immunoradiometric Assay; Keratins; Lung Neoplasms; Male; Middle Aged; Peptide Fragments; Peptides

Keratin 19 is an intermediate filament protein produced by cells of simple epithelia and basal cells of stratified epithelia of different organs. These cell types are associated with important human cancers. We have used the keratin 19 promoter to target the expression of the polyomavirus (Py) large T-antigen, an immortalizing oncogene known to bind to the tumor suppressor retinoblastoma gene product, to epithelial cells. Individuals of the transgenic mouse line K19PyLT-6 developed one or two nodules in one of their lungs. By histology, the nodules were papillary tumors that consisted of nonciliated epithelial cells of the terminal bronchioles. In addition, infiltrates emanating from the nodules were consistent with the development of pulmonary adenocarcinomas. In situ hybridization techniques demonstrated large T-antigen expression in the tumors. Primary cultures were established from a lung tumor dissected from a K19PyLT-6 transgenic mouse. These large T-antigen-expressing cell lines produced the keratin proteins reminiscent of the epithelial origin of the lung tumor. However, further molecular studies indicated that these cell lines did not express Clara cells or pneumocytes markers. s.c. injection of the cell lines into nontransgenic syngeneic mice produced tumors in 2 weeks that resembled malignant pulmonary adenocarcinomas. These animals, which display tumor progression in situ, and the cell lines derived thereof provide a useful system for the study of lung tumorigenesis.

Topics: Adenocarcinoma; Animals; Antigens, Polyomavirus Transforming; Bronchi; Cell Line; Culture Techniques; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Lung Neoplasms; Mice; Mice, Inbred Strains; Mice, Transgenic; Papilloma; Phenotype; Promoter Regions, Genetic; Restriction Mapping; RNA, Viral; Tumor Cells, Cultured

In the current study a comparative analysis of keratin typing and DNA content was carried out in human lung tumors from transthoracic fine needle aspiration biopsies (TFNAB) (18 patients) or from surgically resected tumor tissues (14 patients). According to the cytologic and histologic features, 2 of the 32 tumors were diagnosed as benign tumors, 11 as squamous cell carcinomas, 12 as adenocarcinomas, and 7 as undifferentiated large cell carcinomas. Two cases in the adenocarcinoma and one in the undifferentiated large cell carcinoma groups were pulmonary metastasis or second primary tumors. Malignant cells of tumors which reacted positively with KK8.60 anticytokeratin polypeptides No. 10 and 11 (and hence contain keratinizing cells) displayed diploid DNA content in a flow cytometric assay regardless of their cytologic or histologic appearance. In contrast, all tumors which lacked such positive cells (most of which were defined as adenocarcinomas and undifferentiated tumors) were hyperdiploid. The close correlation between high DNA content and both malignancy and the absence of advanced squamous differentiation (keratinization) suggests that such combined analysis may provide new tools for the cytologic diagnosis and prognosis of lung cancers.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy, Needle; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Cell Cycle; Diploidy; DNA, Neoplasm; Female; Flow Cytometry; G1 Phase; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Resting Phase, Cell Cycle

Adenoid cystic carcinoma (ACC) is a rare but distinctive salivary gland-type malignant neoplasm that arises infrequently as a primary tumor in the lung.. The clinical and pathologic features in 16 cases of primary ACC of the lung were reviewed, and immunohistochemical stains on paraffin sections were performed in 7 cases.. The patients' ages ranged from 29 to 79 years (mean age, 54 years); 11 were men and 5 were women. Clinically, most patients were seen initially with obstructive symptoms, including cough, wheezing, shortness of breath, and hemoptysis. Eight tumors were in the left lung and eight in the right lung. The lesions were treated by pneumonectomy in seven patients, lobectomy in six, and lobectomy plus chemotherapy in two. One patient was treated with chemotherapy alone after undergoing a diagnostic biopsy that revealed advanced disease. Grossly, most lesions were described as endobronchial and measured from 0.9 to 4.0 cm in greatest dimension; two cases, however, showed poorly circumscribed infiltrative tumors. Histologically, three main growth patterns were identified admixed in various proportions: cribriform (cylindromatous), tubular, and solid. Immunohistochemical study in six of seven cases showed a prominent myoepithelial cell component, as evidenced by immunoreactivity for keratin, actin, and S-100 protein in numerous tumor cells. Clinical follow-up ranging from 2 to 15 years in six patients showed that three were alive and well without evidence of recurrence or metastases at 5, 10, and 12 years, respectively, and three were alive with recurrence at 2, 5, and 15 years, respectively. Three other patients died of unrelated conditions at 2, 7, and 9 years, respectively, after diagnosis. Two patients in the study were seen initially with metastatic spread at the time of initial diagnosis and died 2 months and 1 year later with widespread metastases to lymph nodes, liver, spleen, kidney, and bone despite intensive chemotherapy.. Disease stage at the time of diagnosis may play an important role in predicting the clinical outcome of patients with these tumors. Despite their generally slow and indolent growth in other locations, ACC arising in the lung may in certain cases be more aggressive.

Topics: Adult; Aged; Carcinoma, Adenoid Cystic; Combined Modality Therapy; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged

We evaluated the newly produced tumor marker assay kit, "Ball ELSA CYFRA21-1", which detects cytokeratin 19 fragment in the sera of patients with malignancies, especially lung cancers. The assay procedure is simple based on the one-step radioimmunometric assay method. The measured values depend somewhat on incubation temperature and time. Reproducibility and recovery were good. The minimum measurable level was 1 ng/ml. The dilution test was satisfactory. The CYFRA21-1 levels were gradually decreased by repeated freezing and thawing and after seven such exercises its activity dropped to about 70% of that of first assay. The presence of CYFRA21-1 antigen was strongly correlated with TPA antigen and, although some discrepancies could be observed in clinical samples, CYFRA21-1 activity was completely absorbed by anti-TPA antibody-coated beads in one sample. CYFRA21-1 levels of 44 normal controls were below 1.0 ng/ml. Assuming a cut-off value of 2.0 mg/ml, 32.7% of all cases with benign disease had values greater than 2.0 ng/ml. This fell to 21.4% on exclusion of cases of interstitial pulmonary disease. Those with malignant diseases had high CYFRA21-1 levels whether associated with lung cancer or not. The most high positive ratios were observed in squamous cell lung cancer, small cell lung cancer, and uterine cervical cancer. In conclusion, CYFRA21-1 may be a good tumor marker comparable to TPA not only for lung cancer but also other malignancies as well. High false positives for lung cancer, however, were observed in other pulmonary diseases.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Child; Evaluation Studies as Topic; Female; Humans; Immunoradiometric Assay; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasms; Reagent Kits, Diagnostic

The Cyfra 21.1 assay is a newly developed test which measures in serum a fragment of cytokeratin 19. We evaluated this marker in 212 patients with non-small-cell lung cancer (NSCLC), predominantly stage 3a-b and 4, and compared it with three other markers: carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC) and tissue polypeptide antigen (TPA). Sensitivities for Cyfra 21.1, TPA, CEA and SCC (using cut-off levels corresponding to a 95% specificity for benign lung diseases) were 40%, 40%, 42% and 19% respectively. The sensitivity of CEA was significantly higher in patients with adenocarcinomas compared with the other three markers, while the sensitivity of Cyfra 21.1 and TPA was significantly higher in patients with squamous cell carcinomas. The value of Cyfra 21.1 for monitoring disease during chemotherapy could be evaluated in 23 patients with squamous cell carcinomas. When the cases of lead time were included a concordance between clinical evaluations according to WHO response criteria and evaluations according to changes in the marker levels of 74% was found. The criteria defined for marker response were a 65% decrease in the marker level for a partial response and a 40% increase for progressive disease. In particular, increasing levels of this marker indicated usually disease progression. In conclusion, Cyfra 21.1 is a useful serum marker for patients with NSCLC, especially for disease monitoring of patients with squamous cell carcinoma during and after chemotherapy.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Radioimmunoassay

An important role in differentiation and proliferation has been demonstrated for the 20 cytokeratin (CK) polypeptides. The serum of 24 patients with biopsy-proven non-small cell lung cancer (NSCLC) and a similar number of controls was examined for evidence of CK8 and CK18. Using enzyme-linked immunosorbent assay (ELISA), all the control sera were negative, but 9 of the 24 patients were positive (mean 2.62 ng/ml; range 1.4-5.8; P = 0.0036). Western blotting confirmed the results of the ELISA in all cases, and indicated full size CK polypeptides. Advanced stage disease patients were more likely to be seropositive (P = 0.00024). Biopsy specimens showed CK8 expression in all 24 cases by immunochemistry and CK18 in 22 cases. This is the first study to demonstrate that a subgroup of NSCLC patients have intact CK8 and CK18 peptides in their serum, and their detection may correlate with advanced disease.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Cell Death; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Tumor Cells, Cultured

Pulmonary microvascular tumor embolization is a recognized cause of respiratory distress in cancer patients that is rarely diagnosed antemortem. Previous studies with relatively few patients have reported high diagnostic yield using wedged pulmonary artery catheter-derived blood samples to evaluate dyspneic cancer patients for possible microembolization. From 1991 to 1993, 21 cancer patients with respiratory distress of relatively acute onset and varying severity who required pulmonary artery catheterization for hemodynamic monitoring were evaluated using this technique. Pulmonary microvascular cytology (PMC) was interpreted as positive for malignant cells in nine of 21 patients presenting with a range of tumor types, including carcinomas of the breast, colon, and pancreas as well as non-Hodgkin's lymphoma. In 11 patients the PMC was interpreted as negative. One case was considered nondiagnostic. Megakaryocytes, noted in most PMC specimens as well as in several samples of simultaneously drawn peripheral blood, may mimic epithelial tumor cells. Immunocytochemical stains for factor VIII and cytokeratins were used to resolve occasional diagnostic dilemmas. Clinical and/or pathologic follow-up information was available for all patients. Diagnostic accuracy was highest for epithelial malignancies. Two false-negative results occurred in patients with metastatic choriocarcinoma and breast carcinoma. Circulating malignant cells in the peripheral blood of a patient with non-Hodgkin's lymphoma led to one false-positive diagnosis. Benign lymphoid elements in PMC generally have a reactive and variable appearance that should not be misinterpreted as lymphoma. We conclude that PMC is a useful tool in the evaluation of dyspneic cancer patients requiring pulmonary artery catheterization for hemodynamic monitoring and its use potentially avoids additional diagnostic procedures.

Topics: Catheterization, Peripheral; Factor VIII; False Negative Reactions; Hemodynamics; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Megakaryocytes; Microcirculation; Pulmonary Artery; Pulmonary Embolism

Primary lung tumors showing features of salivary gland-type neoplasms are extremely rare.. Eight patients with primary lung neoplasms showing light microscopic and immunohistochemical features of salivary gland-type mixed tumors were studied.. The patients were six women and two men, ages 35-69 years (mean, 52.5 years). The tumors ranged from 2 to 16 cm in greatest diameter. In two patients the lesions presented as polypoid endobronchial lesions obstructing the lumen; in another two patients the lesions were found in close proximity or in continuity with a bronchus; in three patients, the lesions presented as peripheral parenchymatous nodules unrelated to a bronchus; and in one patient, the relationship to the bronchus could not be determined. Histologically, the lesions were biphasic, showing admixtures in varying proportions of epithelial elements containing a predominant myoepithelial cell population with a stromal component containing an abundant myxoid or focally chondroid matrix. Immunohistochemical studies showed strong positivity of the cells in the epithelial component with low molecular weight keratins (CAM 5.2), and to a lesser extent with broad spectrum keratin, actin, and vimentin antibodies. The cells also showed variable reactivity in the epithelial and nonepithelial elements with S-100 protein and glial fibrillary acidic protein. Six tumors were grossly and histologically benign; in two patients, the tumors were larger, locally invasive, and showed more atypical histologic features. All patients were treated with surgical excision. On follow-up, of the six patients with histologically benign-appearing tumors, one was alive and well 6 years after surgery; another died 4 years after surgery of a second unrelated malignancy; one died during the immediate postoperative period of myocardial infarction; and three have been lost to follow-up. In the two patients with histologically atypical lesions, the tumors recurred and metastasized after 2 and 3 years, respectively, with one of them leading to death caused by widespread metastases and superior vena cava syndrome.. Review of the literature and the findings in the current series indicate that salivary gland-type mixed tumors of the lung may present with a spectrum of histologic features and clinical behavior, ranging from benign to frankly malignant, similar to that observed for their salivary gland counterparts. Size of the lesion at the time of presentation, extent of local infiltration, and degree of mitotic activity appear to be the most reliable prognostic features of these tumors.

Topics: Actins; Adenoma, Pleomorphic; Adult; Aged; Female; Glial Fibrillary Acidic Protein; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Mixed Tumor, Malignant; S100 Proteins; Vimentin

Enzyme immunoassay CYFRA 21-1, a novel lung tumor marker presented in serum (upper limit of normal values: 3.5 ng/ml) was measured in 84 patients with lung cancer and 141 patients with benign respiratory diseases. The positive rate for CYFRA 21-1 in primary lung cancer was 70.3% and the false positive rate in respiratory diseases was 24.1%. In respect to the histological types of lung cancer, CYFRA 21-1 was elevated in 85.0% of squamous cell carcinoma and in 64.8% of other cell types of primary lung cancer. Elevated CYFRA 21-1 levels were found in 40% of Stage I-II, in 80.0% of Stage IIIA-B, and in 73.5% of Stage IV. The overall diagnostic efficiency was 53.4% for CYFRA 21-1 and 44.5% for CEA, respectively. Receiver operating characteristic (ROC) curve analysis suggested that CYFRA 21-1 test has an advantage over CEA. From the results of this study, CYFRA 21-1 was more efficient than CEA as primary diagnostic marker in lung cancer.

Topics: Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Prognosis

Metastatic basal cell carcinomas of the skin are rare. We present the cytologic features of a metastatic basal cell carcinoma to the lung diagnosed by fine-needle aspiration biopsy. Cytologic examination revealed syncytial groups of relatively small cells with hyperchromatic, oval to spindle-shaped nuclei having high nuclear to cytoplasmic ratios. Immunocytochemical studies performed on the cell block sections revealed the malignant cells to be positive for cytokeratin (AE1/3) and negative for neuroendocrine markers, [neuron specific enolase (NSE) and chromogranin (Phe-5)]. We reviewed the literature related to metastatic basal cell carcinoma of the skin and discuss risk factors and mechanisms of metastatic spread. In addition, a discussion of the other entities that can enter into the differential diagnosis is presented along with the role of ancillary studies. To the best of our knowledge, we believe this is the first case report of the fine-needle aspiration (FNA) cytology of a basal cell carcinoma metastatic to the lung.

Topics: Biopsy, Needle; Carcinoma, Basal Cell; Female; Humans; Intermediate Filaments; Keratins; Lung Neoplasms; Middle Aged; Skin Neoplasms

Metastases of internal tumors to the oral cavity are unusual and involve in most cases maxilla and mandible. Metastases to the gingival soft tissue are extremely rare.. To report a case of gingival metastasis from undifferentiated carcinoma of the lung.. The lesion was removed and hematoxylin and eosin sections were performed. Immunohistochemical investigations were performed with a standard three-step immunoperoxidase technique on formalin-fixed, paraffin-embedded tissue sections using anti-CEA, anti-S-100, HMB45, and anti-LCA antibodies.. Based on clinicopathologic findings, a diagnosis of metastasis from undifferentiated carcinoma of the lung was established. Further investigations revealed a primary undifferentiated carcinoma of the lung.. Metastasis from internal neoplasms should be considered among other differential diagnoses in the evaluation of gingival tumors. In the present case, onset of oral lesion preceded detection of the primary lung tumor. Complete screening of the patient should therefore follow a diagnosis of gingival neoplasm of unknown origin.

Topics: Aged; Carcinoembryonic Antigen; Carcinoma; Cell Nucleus; Cytoplasm; Gingival Neoplasms; Humans; Keratins; Leukocyte Common Antigens; Lung Neoplasms; Male; S100 Proteins

The levels of the new tumour marker CYFRA 21-1 were assessed in 115 patients with non-small cell lung cancer (NSCLC) and in 45 patients with non-malignant lung disease. Increased levels of CYFRA 21-1 were observed in 47.8%, mostly in patients with squamous cell carcinoma (SCC; 69.1%). Serum CYFRA 21-1 levels were correlated with the stage of SCC type. Positive CYFRA 21-1 levels in patients with SCC were present in 40% of stage I, 61.1% of stage II, and 85.2% of stage III. In addition, SCC patients who presented mediastinal lymph nodes (N2) demonstrated higher serum CYFRA 21-1 levels, compared with patients without mediastinal lymph nodes metastases (N0 or N1). With regard to tumour size, significant difference was observed between T1, T2 and T3. The study also showed that the percentage of patients who survived 18 months with normal preoperative level of CYFRA 21-1 was higher compared with those patients with elevated preoperative levels of this marker, but the differences were not statistically significant.

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Large Cell; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Evaluation Studies as Topic; Female; Humans; Keratins; Lung Diseases; Lung Neoplasms; Lymphatic Metastasis; Male; Neoplasm Staging; Peptide Fragments; Serpins

The diagnostic value of the water-soluble cytokeratin 19 fragment CYFRA 21-1 in lung cancer was assessed in comparison with carcinoembryonic antigen, squamous cell carcinoma antigen, and neuron-specific enolase. The cut-off value, defined as 95% specificity versus a group of 526 patients suffering from benign chest diseases, was set at 3.3 micrograms/l for cytokeratin 19 fragment CYFRA 21-1 (carcinoembryonic antigen: 7.8 micrograms/l, squamous cell carcinoma antigen: 1.9 micrograms/l, neuron-specific enolase: 13.7 micrograms/l). Elevated pretreatment cytokeratin 19 fragment CYFRA 21-1 concentrations were recorded: in 112 of 244 (46%) patients with all histological types of lung cancer (carcinoembryonic antigen: 32%, squamous cell carcinoma antigen: 25%, neuron-specific enolase: 28%), in 89 of 177 (50%) patients with non-small cell lung cancer (carcinoembryonic antigen: 33%, squamous cell carcinoma antigen: 24%, neuron-specific enolase: 12%), in 47 of 81 (58%) patients with squamous cell carcinoma (carcinoembryonic antigen: 23%, squamous cell carcinoma antigen: 32%, neuron-specific enolase: 14%), in 27 of 63 (42%) patients with adenocarcinoma (carcinoembryonic antigen: 44%, squamous cell carcinoma antigen: 14%, neuron-specific enolase: 9%), in 15 of 33 (45%) patients with other non-small cell lung cancer (carcinoembryonic antigen: 36%, squamous cell carcinoma antigen: 24%, neuron-specific enolase: 14%), and in 20 of 55 (36%) patients with small cell lung cancer (carcinoembryonic antigen: 32%, neuron-specific enolase: 77%). Three of 12 patients with undefined histological type showed cytokeratin 19 fragment CYFRA 21-1 elevations. The best performance in terms of sensitivity and diagnostic accuracy was attained with the cytokeratin 19 fragment CYFRA 21-1 test in squamous cell carcinoma. In small cell lung cancer neuron-specific enolase was confirmed to be superior to the other markers. Cytokeratin 19 fragment CYFRA 21-1 concentrations increased with the extent of the malignant disease in non-small cell lung cancer. The positivity rate of cytokeratin 19 fragment CYFRA 21-1 in tumour stage TNM I was only 23% (carcinoembryonic antigen: 23%, squamous cell carcinoma antigen: 14%), i.e. the markers under study cannot be used for the diagnosis of early stage disease. Cytokeratin 19 fragment CYFRA 21-1 differentiated significantly between squamous cell carcinoma and the other histological types (p < 0.01). In addition, cytokeratin 19 fragment CYFRA 21-1 distinguishe

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; International Cooperation; Keratins; Lung Neoplasms; Male; Peptide Fragments; Phosphopyruvate Hydratase; Sensitivity and Specificity; Serpins

We used RNA-RNA in situ hybridization to study expression of the human CC10 gene in morphologically normal and atypical areas of 32 non-neoplastic lung specimens resected from 26 non-small cell lung cancer patients. We scored strong, moderate or weak levels of CC10 mRNA expression in 3 distinct lung compartments. In morphologically normal lungs, strong and moderate levels of CC10 mRNA were observed in bronchioli and bronchi, respectively, but the expression was rarely observed in the alveolar region. Distinct alterations in CC10 mRNA expression were noted in specific histologic abnormalities within bronchi and the alveolar region. CC10 hybridization signal decreased markedly in bronchi containing diffuse goblet cell hyperplasia or squamous metaplasia, while CC10 mRNA expression remained unchanged in bronchi with basal cell hyperplasia or focal goblet cell hyperplasia. Bronchiolar CC10 mRNA levels remained unchanged in sections containing abnormalities elsewhere. Interestingly, in alveoli with bronchiolization of the alveoli, high levels of CC10 mRNA were observed. These regions also contained strongly stained keratin 14-positive cells, which may indicate a concurrent metaplastic process. In lungs with morphologic atypias, no correlation was found between abnormalities detected in bronchi and alveoli from the same lung. A comparison of mRNA expression and clinicopathologic features demonstrated that the amount of histologic abnormalities increased with smoking history (pack years); however, no correlation between CC10 mRNA expression and sex, age or smoking history was found.

Topics: Age Factors; Carcinoma, Non-Small-Cell Lung; Female; Gene Expression; Humans; In Situ Hybridization; Keratins; Lung Neoplasms; Male; Mesothelioma; Proteins; RNA, Messenger; Sex Factors; Uteroglobin

The present study was designed to determine whether CYFRA 21-1, measuring cytokeratin 19, could be a specific and sensitive tumour marker for non-small cell lung cancer (NSCLC). Serum measurements were made at diagnosis in 2250 patient samples by an immunoradiometric "sandwich type" assay, using two cytokeratin 19 specific monoclonal antibodies. Among healthy individuals (n = 711) and patients with benign lung disease (n = 546), 95 percentiles were 1.2 and 2.95 ng/ml, respectively. Cumulative distribution analysis curves were established. From these data, 3.3 ng/ml gave 96% specificity. Using this cutoff, the sensitivity for small cell lung cancer was 16% (n = 74) compared to 41% for NSCLC (n = 547). In histological sub-groups, sensitivity was 57% for squamous cell lung cancer, 34% for undifferentiated large cell carcinoma and 27% for adenocarcinoma, the level of CYFRA 21-1 was correlated with tumour size and UICC stage. In squamous cell lung cancer, the sensitivity of the squamous cell carcinoma marker was 30%, 25% for carcinoembryonic antigen and 46% for tissue polypeptide antigen, using the same series of samples and cutoffs defined at 96% specificity. In conclusion, CYFRA 21-1 is a sensitive tumour marker for NSCLC, especially squamous cell lung cancer.

Topics: Adolescent; Adult; Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Diseases; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Neoplasms; Peptide Fragments; Retrospective Studies; Sensitivity and Specificity

The aim of the study is to estimate the new tumor marker CYFRA 21-1 in non-small cell lung cancer (NSCLC) patients and comparison of this results with SCC-Ag. The investigation was carried out on 115 NSCLC patients (55 with squamous cell, 35 with adenocarcinoma, 25 with large cell) qualified for surgical treatment and in 48 nonmalignant lung diseases patients. CYFRA 21-1 was determined by the means of IRMA method (CIS bio international--GIF-SUR-Yvette, France) and SCC-Ag--MEIA method (IMx system Abbott). Elevated levels of CYFRA 21-1 were obtained in 48.7% and SCC-Ag in 39.1%. Elevated levels of examined markers most frequently occurred in squamous cell type (SCC). It was found out that CYFRA 21-1 dependent on: a) SCC stage (I-40%, II-61.1%, III-85.2%), b) tumor size (T1-38.4%, T2-73.1%, T3-87.5%), c) mediastinal lymph nodes metastases (No and N1-53.8% and N2-86.9%). Similar correlations were not observed in SCC-Ag examination. Simultaneous determination of CYFRA 21-1 and SCC-Ag showed minimal sensitivity increase from 48.7% to 52.1% in NSCLC and from 69.1% to 70.1% in SCC and decrease of specificity from 95.8% to 85.4%. To sum up, determination of CYFRA 21-1 in NSCLC patients (especially in SCC patients) is useful in diagnosis and clinical stage determination.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma; Carcinoma, Non-Small-Cell Lung; Humans; Keratins; Lung Neoplasms; Middle Aged; Neoplasm Staging; Peptide Fragments; Sensitivity and Specificity; Serpins

Topics: Antibody Specificity; Humans; Keratins; Lung Neoplasms; Peptides; Tissue Polypeptide Antigen

Serum samples from 137 lung cancer patients were examined by RIA to evaluate the clinical efficacy of the new tumor marker, CYFRA 21-1, which could identify the soluble fragment of cytokeratin 19. The cut-off value was determined to be 2.2 ng/ml according to the receiver operating characteristic curve. The sensitivity, specificity and accuracy of the RIA for CYFRA 21-1 were 57.7, 91.9 and 64.9%, respectively. The serum concentration of CYFRA 21-1 and the sensitivity of the assay increased as the disease progressed. Histologically, the sensitivity was highest for squamous cell carcinomas (SQ) (76.5%) in comparison with adenocarcinomas (47.8%) and small cell lung cancers (42.1%) (P < 0.01, P < 0.05, respectively). The sensitivities for SQ were 60.0, 83.3, 80.0 and 100% at stages I, II, III and IV, respectively. When compared with CEA (45.3%) and squamous cell carcinoma related antigen (SCC) (22.6%) in all lung carcinomas, CYFRA 21-1 showed the highest sensitivity (57.7%), (P < 0.05, P < 0.01, respectively). In SQ, the sensitivity of the CYFRA 21-1 RIA was significantly higher than that of the assay for SCC (47.1%) (P < 0.05). In patients with adenocarcinomas, the sensitivity of the CYFRA 21-1 assay was almost the same as that for CEA (49.3%). In a combination of CYFRA 21-1 and CEA for non-small cell lung cancers (NSCLC), the sensitivity and accuracy increased to 75.4 and 78.1%, respectively, although the specificity decreased to 86.5%. It is concluded that CYFRA 21-1 could replace SCC, a less satisfactory tumor marker, for SQ of the lung. The potentiality of the combination of CYFRA 21-1 and CEA for NSCLC should be estimated using larger samples in the near future.

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Radioimmunoassay; Sensitivity and Specificity

A 36-year-old Russian patient had been exposed to severe nuclear radiation during the Tschernobyl accident and developed Cushing's syndrome in September 1991. After bilateral adrenalectomy a small centrally localized lung tumor in the left segment S6 was diagnosed, and the syndrome was correlated to primary lung cancer. Thoracotomy and resection of the segments S6 revealed a primary ACTH-producing lung tumor of the cell type of a paraganglioma. The patient lost his symptoms after curative tumor resection. The case is discussed under the aspects of tumor etiology, its clinical course and immunohistochemical findings.

Topics: Adrenocorticotropic Hormone; Adult; Cushing Syndrome; Humans; Keratins; Lung Neoplasms; Male; Neoplasms, Radiation-Induced; Paraganglioma; Phosphopyruvate Hydratase; Power Plants; Radioactive Hazard Release; Tumor Suppressor Protein p53; Ukraine; Vimentin

Adenocarcinomas of uncertain origin are a frequent problem for surgical pathologists. To determine the utility of immunostaining for cytokeratin 7 and cytokeratin 20 in the separation of pulmonary adenocarcinomas from colonic adenocarcinomas, we studied routinely processed, formalin-fixed tissue from 151 of these tumors using commercially available monoclonal antibodies and an avidin-biotin immunohistochemical technique. Used alone, neither cytokeratin 7 immunostaining or cytokeratin 20 immunostaining reliably separated these tumors. However, the immunophenotype of cytokeratin 7 positive/cytokeratin 20 negative was seen in 86% of the pulmonary adenocarcinomas, and in 0% of the colonic adenocarcinomas. Conversely, the cytokeratin 7-negative/cytokeratin 20-positive immunophenotype was seen in 77% of the colonic carcinomas, and in 0% of the pulmonary tumors. In conclusion, cytokeratin 7/cytokeratin 20 immunostaining patterns may be helpful in separating pulmonary adenocarcinomas from colonic adenocarcinomas.

Topics: Adenocarcinoma; Colonic Neoplasms; Diagnosis, Differential; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratins; Lung Neoplasms

Soft agar clonogenic assays are considered to be a standard method for measuring tumour cell radiosensitivity and it has been widely reported that fibroblast contamination does not occur. We report here that human fibroblasts can proliferate to form colonies in a modified form of the Courtenay-Mills soft agar clonogenic assay. It was observed that early passage skin fibroblasts could form colonies in soft agar, although the plating efficiencies were reduced compared with growth on plastic. It was demonstrated that normal lung could proliferate in agar with similar plating efficiencies to fresh tumours and that fibroblastic cells were present in these cultures. Characterisation of primary lung tumour cultures also showed that fibroblastic cells were present in these cultures. Characterisation of primary lung tumour cultures also showed that fibroblastic cells were present which lacked epithelial features and which resembled closely the cells found in cultures of normal lung. This is an important finding for workers using soft agar assays to culture human tumour cells and is of interest in understanding the processes of normal growth control of human fibroblasts.

Topics: Agar; Cell Division; Cells, Cultured; Culture Media; Epithelial Cells; Epithelium; Fibroblasts; Humans; Keratins; Lung; Lung Neoplasms; Microscopy, Electron; Radiation Tolerance; Skin; Tumor Cells, Cultured; Tumor Stem Cell Assay

We evaluated the diagnostic usefulness of measurement of the soluble cytokeratin 19 fragment, a new tumor marker, in 391 patients with lung cancer and in 424 patients with benign lung diseases. Serum concentrations of cytokeratin 19 fragment were measured by a sandwich ELISA (CYFRA). The cut-off value was defined as 3.5 ng/ml, which is associated with a specificity of 85% for benign lung diseases. CYFRA had a high sensitivity (57.5%) in all subjects with lung carcinoma, and had a higher sensitivity for squamous cell carcinoma (73.1%, n = 141) than squamous cell carcinoma-related antigen (61.0%). CYFRA was associated with a relatively high sensitivity (42.1%) in early-stage squamous cell carcinoma (stage I, based on the classification of the Japan Lung Cancer Society), but the CYFRA titer was higher in advanced squamous cell carcinoma than in early-stage squamous cell carcinoma. Our findings suggest that CYFRA is potentially useful for diagnosis and monitoring of lung carcinoma, especially for squamous cell carcinoma.

Topics: Biomarkers, Tumor; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Lung Neoplasms; Male; Peptide Fragments; Reagent Kits, Diagnostic; Sensitivity and Specificity

Topics: Humans; Keratins; Lung Diseases; Lung Neoplasms; Reagent Kits, Diagnostic; Reference Values; Sensitivity and Specificity

Serum level of cytokeratin 19 fragment; which is a tumor associated antigen of lung cancer; was detected by the radioimmunometric assay. Reliability for clinical laboratory measurement was investigated. Detective range of cytokeratin 19 fragment was 0-50 ng/ml and the results including reproducibility, linearity of the data from diluted standard samples, was good enough for laboratory practice use. Although, influence of high concentration of CA19-9 over cytokeratin 19 fragment assay was suspected, there was no correlation between amount of added the antigen to data of cytokeratin 19 fragment. So direct cross reactivity of cytokeratin 19 fragment antibody to CA19-9 seemed less possible. As inhibitory effect of samples which contained over 100 ng/ml of cytokeratin 19 fragment on assay was observed due to antigen excess, evaluation by diluted sample was necessary for the samples that showed over detective limit.

Topics: Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Humans; Keratins; Lung Neoplasms; Radioimmunoassay; Reagent Kits, Diagnostic; Reproducibility of Results

Recently CYFRA 21-1, a new tumour marker measuring a fragment of cytokeratin 19, was introduced and proved to be suitable for the follow-up care and monitoring of the therapy of non-small cell lung carcinomas, especially squamous cell carcinomas of the lung. Besides CYFRA 21-1, there are two other tumour markers available, called tissue polypeptide antigen (TPA) and tissue polypeptide specific antigen (TPS), which also measure different cytokeratins in serum. In a retrospective study we investigated the clinical significance of these 3 cytokeratin markers in lung cancer compared with carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC) and neuron-specific enolase (NSE). We investigated the sera of 50 healthy persons, 273 patients with various benign diseases and 218 patients with histologically proven lung cancer. In a first step the specificity versus benign diseases of the lung was established for all the markers, and was fixed at 95%. Then the single and combined sensitivities were calculated. CYFRA 21-1 proved to possess the highest sensitivity in lung cancer in general (61%), in non-small cell lung carcinomas (64%), in squamous cell carcinomas (79%), in adenocarcinomas (54%) and in large cell carcinomas (65%). In small cell lung carcinomas, neuron-specific enolase proved again to be the marker of first choice (55%). Combined determinations proved clearly increased sensitivity only for large cell carcinomas (CYFRA 21-1 + TPA: 77%) and for small cell lung carcinomas (CYFRA 21-1 + NSE: 62%).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Female; Gastrointestinal Diseases; Genital Diseases, Female; Humans; Keratins; Lung Diseases; Lung Neoplasms; Male; Peptides; Reference Values; Retrospective Studies; Sensitivity and Specificity; Tissue Polypeptide Antigen

Cytokeratin 19 is a subunit of cytokeratin intermediate filament expressed in simple epithelia and their malignant counterparts. Therefore, it is expressed by respiratory epithelium cells and has been detected in lung cancer specimens. An immunoradiometric assay was used to detect a fragment of the cytokeratin 19, referred to as CYFRA 21-1, in the serum of 165 patients with histologically proved lung cancer (128 non-small cell and 37 small cell lung cancers). This prospective study was conducted to evaluate the reliability of this immunoradiometric assay and to identify the relationship between serum CYFRA 21-1 and different features of lung cancer including prognosis. The minimal detectable concentration detected by this assay was 0.06 ng/ml. The reliability of the immunoradiometric assay was demonstrated by the linear relationship between CYFRA 21-1 measurement and dilution of the serum, the reproducibility of the dosage in intraassay and interassay, and the high sensitivity of the method in discriminating low CYFRA 21-1 concentrations. Using a threshold of 3.6 ng/ml, sensitivity and specificity were 0.52 and 0.87, respectively. The sensitivity of the marker was highest in squamous cell carcinoma and lowest in small cell carcinoma. In non-small cell lung cancer patients, the marker varied significantly according to both stage of the disease (Kruskal-Wallis, 13.7; P < 0.005) and performance status (Kruskal-Wallis, 9.16; P < 0.05) inasmuch as a high serum CYFRA 21-1 level was associated with advanced stages, mediastinal lymph nodes, and poor performance status. Consequently, the marker was significantly lower in patients who were operated upon when compared with unresectable ones. Lung cancer patients with serum CYFRA 21-1 over 3.6 ng/ml proved to have a significantly shorter overall survival than those with a normal serum level (log rank, P = 0.007; Wilcoxon, P = 0.001). The negative prognostic effect of CYFRA 21-1 was highly significant in squamous cell carcinomas whereas it was nonsignificant for the other histologies. In Cox's model analysis, performance status, stage grouping, and CYFRA 21-1 were the only significant determinants of survival. This study supports the use of the serum fragment of cytokeratin subunit 19 CYFRA 21-1 as an independent prognostic marker of squamous cell carcinoma of the lung.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Evaluation Studies as Topic; Female; Genetic Variation; Humans; Immunoradiometric Assay; Keratins; Lung Neoplasms; Macromolecular Substances; Male; Middle Aged; Peptide Fragments; Prospective Studies; Sensitivity and Specificity

The immunoreactivity of OV-TL 12/30, a monoclonal antibody to keratin 7 was investigated on paraffin-embedded human lung cancer tissues of 61 patients. A modified AEC-immunoperoxidase method with pepsin pre-digestion was used. In normal lung tissue keratin 7 was found in bronchial and bronchiolar epithelium, pneumocytes and compound glands. Squamous metaplasia of the bronchial tree was negative. All 24 squamous cell carcinomas were negative irrespective of grade of differentiation. All differentiation grades of 20 adenocarcinomas including bronchioalveolar carcinomas were positive. Since six large cell anaplastic carcinomas did not react with keratin 7 antibody these tumours are considered to be of squamous cell rather than adenocarcinomatous origin. Small cell anaplastic carcinomas were negative in 10 of 11 cases. Our study demonstrates that this keratin 7 antibody is useful in differentiating between squamous cell carcinoma and adenocarcinoma of the lung and it may be particularly useful in making the correct diagnosis in small lung biopsy specimens.

Topics: Antibodies, Monoclonal; Carcinoma; Humans; Immunoenzyme Techniques; Keratins; Lung; Lung Neoplasms

Present diagnostic techniques do not allow the detection of early metastatic spread of tumor cells, although this spread largely determines the clinical course of patients with small primary cancers. By use of monoclonal antibody CK2 to the epithelial cytokeratin component number 18 (CK18), individual disseminated carcinoma cells present in bone marrow of cancer patients can now be identified (G. Schlimok, I. Funke, B. Holzmann, G. Göttlinger, G. Schmidt, H. Häuser, S. Swierkot, H. H. Warnecke, B. Schneider, H. Koprowski, and G. Riethmüller, Proc. Natl. Acad. Sci. USA, 84: 8672-8676, 1987; F. Lindemann, G. Schlimok, P. Dirschedl, J. Witte, and G. Riethmüller, Lancet, 340: 685-689, 1992). In the present study, we applied this approach to patients with operable non-small cell lung cancer. CK18 was expressed on 84 of 88 (95.5%) primary adenocarcinomas and squamous cell carcinomas. Irrespective of primary tumor histology, single aspirates of iliac bone marrow from 18 of 82 (21.9%) lung cancer patients exhibited between 1 and 531 CK18+ cells/4 x 10(5) nucleated marrow cells. The specificity of our assay is underlined by the small rate of "false-positive" cells being observed in only 2 of 117 (1.7%) marrow samples from control patients with no evidence for an epithelial malignancy at the time of aspiration. Comparison with established risk factors demonstrated positive correlations (P < 0.05) between the size and histological grade of the primary carcinoma and cytokeratin positivity in iliac bone marrow. In contrast, the association with the metastatic involvement of regional lymph nodes was only weak (P = 0.09). Following a median observation period of 13 months, patients who displayed cytokeratin-positive cells in iliac bone marrow at the time of primary surgery relapsed more frequently as compared to patients with a negative marrow finding (66.7 versus 36.6%; P < 0.05). This difference was even more pronounced by comparing the rates of manifest skeleton metastasis observed in both groups (26.7 versus 2.4%; P < 0.005). Finally, colabeling of CK18+ cells in marrow with monoclonal antibodies to proliferation-associated markers, such as the nucleolar antigen p120 or the tyrosine kinase receptor erbB2, exemplified the oncogenic capacity of CK18+ micrometastatic cells. In conclusion, CK18+ cells present in the bone marrow of patients with apparently operable non-small cell lung cancer exhibit the potential to form solid metastases. Therefore, the approach presente

Topics: Adult; Aged; Aged, 80 and over; Bone Marrow; Bone Marrow Diseases; Carcinoma, Non-Small-Cell Lung; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Proto-Oncogene Proteins; Receptor, ErbB-2; Risk Factors

Accurate assessment of the presence and absence of tumor in the regional lymph nodes is critical in assessment of prognosis for patients with lung cancer. Development of sensitive immunohistochemical techniques and specific monoclonal antibodies has increased our capacity, in melanoma and breast cancer, to detect small groups of tumor cells or even single tumor cells in lymph nodes that appear to be tumor free in conventionally stained sections.. This retrospective study was designed to assess whether use of a polyclonal antikeratin reagent in immunohistochemical analysis offers any advantage over conventional histopathology in detection of regional lymph node metastases in non-small-cell lung cancer.. Paraffin-embedded tissue sections from regional lymph nodes of 65 patients with non-small-cell lung cancer were studied. We examined tissue from 588 nodes of 60 patients with a diagnosis of disease confined to the lung and from 72 nodes of five patients with a diagnosis of metastasis to some nodes. A polyclonal antikeratin antibody was applied to the lymph node tissue sections, using the avidin-biotin complex immunoperoxidase technique.. Single tumor cells and small clusters of tumor cells (occult micrometastases) not visible on routine evaluation were readily detected in 38 (63%) of the 60 patients whose nodes appeared to be negative on examination of hematoxylin-eosin-stained slides. In the five patients with a diagnosis of node-positive non-small-cell lung cancer, five (10%) of 51 nodes that were tumor free on conventional examination contained metastases. Metastatic tumor cells were most often located in the subcapsular or medullary sinuses. The lymph nodes that contained occult tumor cells were those located nearest to the tumor, mainly in peribronchial and hilar locations. The median survival of patients with occult metastases (1977 days) was shorter than that of patients whose nodes contained no tumor (2456 days) but was longer than that of patients whose nodes contained metastases detectable on hematoxylin-eosin-stained slides (927 days).. Our results suggest that, in patients with non-small-cell lung cancer, metastatic involvement of regional lymph nodes is more frequent than was previously determined by the conventional histologic method and substantially more frequent than in other tumor types, such as melanoma and breast cancer.. The high frequency of occult nodal metastases in non-small-cell lung cancer makes it clear that, without immunohistochemistry, disease is understaged in many patients. Therefore, it seems essential that immunohistochemical evaluation of the lymph nodes be undertaken in clinical trials.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Middle Aged; Retrospective Studies

The nuclear DNA contents of atypical mesothelial cells from five patients who had an eosinophilic pleural effusion (EPE) were studied by the use of DAPI (4',6-diamidino-2-phenylindole dihydrochloride) DNA staining. Analysis of the nuclear DNA content revealed a polyploid pattern, with a major peak in the tetraploid region. Using an immunocytochemical technique, the atypical mesothelial cells showed a positive reaction for cytokeratin. In contrast carcinoembryonic antigen (CEA) was always negative in these cells. It is suggested that the atypical mesothelial cells with EPE had a higher rate of proliferation than did the normal mesothelial cells.

Topics: Adult; Aged; Biomarkers; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Cell Division; Cell Nucleus; Crohn Disease; DNA; Eosinophilia; Epithelium; Female; Humans; Indoles; Keratins; Lung Neoplasms; Male; Mediastinal Cyst; Membrane Glycoproteins; Mucin-1; Pleural Effusion; Pneumonia; Pneumothorax; Polyploidy

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged

We have established two cisplatin-resistant human lung squamous-carcinoma cell lines, PC10-B3 and PC10-E5, from their original cell line PC10. To discover which proteins are associated with cisplatin resistance, we carried out a two-dimensional gel electrophoresis to analyze differences in protein alteration between PC10, PC10-B3 and PC10-E5. A protein spot M(r) 50 kDa, pI5.3, was reduced markedly and a spot M(r) 50 kDa, pI4.9 was increased when PC10-B3 and PC10-E5 were compared with PC10. A spot M(r) 58 kDa, pI5.8 newly appeared only in PC10-E5. Cell fractionation showed that the M(r) 50 kDa, pI5.3 (p50-5.3) and the M(r) 50 kDa, pI4.9 fell within the nuclear fraction, while the M(r) 58 kDa, pI5.8 was found among the cytosol and microsomal fractions. Microsequencing after in situ digestion of the dramatically reduced spot p50-5.3 revealed that it was identical to 50 kDa, type I keratin (K14). Moreover, a retinoic acid-mediated K14 reduction was concomitant with a 4.0-fold increase in cisplatin resistance in PC10. Our report is the first to suggest the possible association of marked K14 reduction and cisplatin resistance in PC10-B3 and PC10-E5.

Topics: Amino Acid Sequence; Carcinoma, Squamous Cell; Cell Survival; Cisplatin; Clone Cells; Drug Resistance; Humans; Keratins; Lung Neoplasms; Molecular Sequence Data; Tumor Cells, Cultured

The immunohistochemical expression patterns of cytokeratins 8 and 18 and vimentin were examined in frozen sections of 120 human mucosal squamous cell carcinomas with special emphasis on the topological distribution in the tumour. This was done in order to evaluate in squamous cell carcinoma a particular expression pattern observed recently by us in transitional cell carcinoma of the urinary tract and designated as an 'interface phenomenon'. This phenomenon implying maximum expression of cytokeratins 8 and 18 at the tumour front, and to a lesser extent also in areas of intratumorous stroma contact, was also found in about 50 per cent of the squamous cell carcinomas examined. It was even found for vimentin, which contrasted with transitional cell carcinoma. The percentages of occurrence of the phenomenon varied for the different sites of origin of the tumour. Tumour grade did not influence the results. These findings further support the idea that invasive carcinoma cells interacting with the stromal micro-environment display a characteristic intermediate filament phenotype that deviates from the pattern expected on the basis of their direction of differentiation. These changes might reflect phenotype involved in invasive, migrating, and proliferating activities.

Topics: Carcinoma, Squamous Cell; Esophageal Neoplasms; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Neoplasm Invasiveness; Neoplasm Proteins; Vimentin; Vulvar Neoplasms

It is known that cytokeratin 19 is particularly abundant in carcinoma of the lung.. A sandwich enzyme-linked immunosorbent assay called CYFRA 21-1 was, therefore, developed to detect soluble cytokeratin 19 fragments in serum using two specific monoclonal antibodies (Ks 19.1 and BM 19.21). The authors investigated the clinical significance of this new marker compared with the established markers carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), neuron-specific enolase (NSE), carbohydrate antigen (CA) 19-9, CA 125, CA 15-3, CA 72-4, alpha-fetoprotein, and prostate-specific antigen in a pilot study on 1741 serum samples from patients with various benign and malignant diseases.. Postulating a specificity of 95% versus benign diseases of the lung, the diagnostic sensitivity of CYFRA 21-1 in lung cancer (independent of histologic type) at primary diagnosis was superior (47%) to CEA (27%), SCC (15%), and NSE (16%). Especially in squamous cell carcinomas of the lung, the true-positive test results were much higher for CYFRA 21-1 (60%) than for CEA (18%) or SCC (31%).. In small cell lung carcinomas, NSE was confirmed as the marker of first choice. For all of the other solid tumors investigated, CYFRA 21-1 showed no better profile of specificity and sensitivity than the established markers.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Lung Neoplasms; Male; Neoplasms; Phosphopyruvate Hydratase; Pilot Projects; Retrospective Studies; Sensitivity and Specificity; Serpins

The expression of cytokeratins (CK) 1, 4, 5/6, 8, 13, 18, 19 and 20 and involucrin in 42 cases of squamous cell carcinomas from various locations was examined. The tumours expressed CK5/6 in 55%, CK8 in 76%, CK13 in 43% and CK19 in 95% of cases. The CK5/6-positive primary tumours were from uterine cervix, head and neck, lung, skin, oesophagus and urinary bladder, and the CK13-positive primary tumours were from uterine cervix, lung and vulva. Metastatic squamous cell carcinomas from head and neck more frequently expressed CK5/6 and 13, 7/7 (100%) and 6/7 (86%) compared with 3/5 (60%) and 0/5 (0%) in the primary squamous cell carcinomas. Few cases were CK1, CK4 and CK18 immunoreactive. CK20 immunoreactivity was not observed. Involucrin was expressed in 71% of tumours, and most of the involucrin-positive cells were located at the central parts of tumour cell clusters except for one case in which the peripheral cells around tumour cell clusters were positive. Thus, expression of the so-called simple epithelial markers CK8 and CK19 occurs in the majority of squamous cell carcinomas. The absence of CK20 immunoreactivity may be helpful in differential diagnosis.

Topics: Carcinoma, Squamous Cell; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Mouth Neoplasms; Protein Precursors; Skin Neoplasms; Urinary Bladder Neoplasms; Uterine Neoplasms

Cytokeratins are intermediate filaments of the cytoskeleton that are expressed by bronchial epithelium and its neoplastic counterpart, lung cancer. A new immunoradiometric assay referred to as CYFRA 21-1 makes it possible to titrate in the serum a cytokeratin 19 fragment. This study deals with the sensitivity, specificity and applicability of this serum marker in squamous cell carcinoma. Sera from non malignant pulmonary diseases were taken as controls. In comparison with carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC T-A4) and neuron specific enolase (NSE), CYFRA 21-1 was the most accurate marker. The area under the CYFRA 21-1 ROC curve was significantly greater than those of CEA, SCC T-A4 and NSE. Using a 3.6 ng/ml threshold, as determined by the ROC curve, CYFRA 21-1 was significantly correlated with tumor mass.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Phosphopyruvate Hydratase; Radioimmunoassay; Sensitivity and Specificity; Serpins

The proliferative cystic squamous cell lesion inducible by inhalation of particulate matter in the rat lung is characterized by the formation of keratin-filled cystic cavities of various sizes bordered by multiple layers of keratinizing squamous epithelial cells. The current investigation is primarily concerned with two points. One is whether the cells participating in this particular cystic configuration can recapitulate their specific in vivo morphogenetic behavior also in the in vitro circumstances. The other is whether these squamous epithelial cells are neoplastic in nature. Although the currently adopted cell culture system was two-dimensional, the specific morphogenetic pattern was reproduced in vitro in a corresponding manner by the squamous cells derived from the aforementioned rat lung lesions. Exposure of these cells to 12-O-tetradecanoyl-phorbol-13-acetate (TPA) at 100 ng/ml induced a marked polymorphism in them and also stimulated their keratinization. In soft agar culture, both TPA-exposed and unexposed cells developed colonies larger than 0.05 mm in diameter with an incidence of 0.73% and 1.12%, respectively. The lower incidence in TPA-exposed cultures can be interpreted in terms of TPA stimulation of keratinization. However, colonies larger than 0.1 mm in diameter were also developed by the TPA-exposed cells, indicating the presence of a cell population responsive to promotional effects of TPA. These results imply that the cells involved in the formation of cystic squamous epithelial cell lesions are already initiated and possess a potential for autonomous benign growth.

Topics: Animals; Cell Division; Cells, Cultured; Cysts; Female; Keratins; Lung; Lung Neoplasms; Rats; Tetradecanoylphorbol Acetate

We developed a new and automated assay for the detection of lung cancer associated cytokeratin 19 fragments in patients' sera/plasma. This new tumour marker assay CYFRA 21-1 was evaluated in technical and clinical studies using the multibatch analysers ES 300 and ES 600 from Boehringer Mannheim GmbH. The analytical performance was shown to be excellent. The clinical data from 2,037 patients demonstrate that for non-small-cell lung carcinoma CYFRA 21-1 has a higher diagnostic sensitivity compared to the established markers. Mainly for squamous cell carcinoma CYFRA 21-1 was superior (60%) to CEA (18%) or SCC (31%).

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Lung Neoplasms; Male; Ovarian Neoplasms; Peptide Fragments; Sensitivity and Specificity; Stomach Neoplasms

A cell line of human lung large cell carcinoma (LCC) was established directly from the metastatic skin tumor tissue. The clinical course of the patient who carried this carcinoma was peculiar; generalized lymphadenopathy, histologically resembling Hodgkin's disease, was found as the first clinical symptom. The lung tumor was not discovered until the time of autopsy. This cell line (KaMi) grew adherent to culture vessels with the population doubling time of 20.6h, formed colonies in soft agars with efficiency of 22.6%, and formed tumors in athymic nude mice. The authenticity of KaMi was confirmed by chromosomal analysis and isoenzyme patterns. KaMi cells bore a strong resemblance to the original tumor cells which were composed of small spindle cells, large polygonal cells, and multinucleated giant cells. Immunohistochemically, KaMi cells showed a weak tendency to differentiate to squamous cells, and these immunohistochemical reactivities were almost compatible to those of the original tumor cells, but ultrastructurally, KaMi cells were more immature than the original ones. Treatment with several reagents could not augment a differentiation of KaMi cells. Cytokeratin profiles showed a tendency of squamous cell differentiation. KaMi cells may aid in elucidating the pathogenesis and biology of LCC and its relationship to other lung tumors.

Topics: Animals; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Cell Differentiation; Chromosome Banding; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Mice; Mice, Nude; Microscopy, Electron; Middle Aged; Neoplasm Transplantation; Tumor Cells, Cultured; Tumor Stem Cell Assay

We describe the microscopic, histochemical, immunohistochemical, and ultrastructural features of hundreds of neuroendocrine tumorlets occurring within a pulmonary lobe severely scarred by intralobar sequestration in a nonsmoking 49-year-old white man. To our knowledge, there have thus far been no descriptions or detailed analyses of neuroendocrine tumorlets arising within a pulmonary sequestration. The neuroendocrine tumorlets appeared in the form of minute aggregates--mostly microscopic, up to a maximum of 0.3 cm in greatest diameter--of small round and short spindle-shaped cells. They were organized in compact nests of fascicles and were supplied with round or elongated euchromatic nuclei and scant weakly eosinophilic cytoplasm. The neuroendocrine tumorlets were clustered around diseased bronchioles or embedded in a fibrotic pulmonary parenchyma with a distinctive infiltrative appearance. Sometimes they lay near an artery channel without an identifiable bronchiole or herniated into distal airways. Most of the neuroendocrine tumorlets were strongly argyrophilic on Grimelius staining. Immunohistochemically, there was reactivity for markers of epithelial and neuroendocrine differentiation together with evidence of orthotopic production of calcitonin, serotonin, and gastrin-releasing peptide and ectopic production of vasoactive intestinal peptide. Ultrastructurally, most of the neuroendocrine cells showed 100- to 120-nm dense-core membrane-bound secretory granules; mucus secretory cells were also present. We prefer the term neuroendocrine tumorlets over the generally used term carcinoid tumorlets, because the nature of these lesions is undefined and the relationship with neuroendocrine pulmonary neoplasms is not yet established.

Topics: Calcitonin; Cell Nucleus; Chromogranins; Cytoplasm; Cytoplasmic Granules; Gastrin-Releasing Peptide; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Microscopy, Electron; Middle Aged; Neurosecretory Systems; Peptides; Phosphopyruvate Hydratase; S100 Proteins; Serotonin; Staining and Labeling; Vasoactive Intestinal Peptide

Lung carcinomas were studied immunohistochemically and the results were related to type of tissue sample (bronchoscopic biopsies, surgical specimens, autopsies). All cytokeratins (CAM 5.2, PKK-1, AE1/AE3) reacted with virtually all adenocarcinomas, most squamous, and 65% of the large cell carcinomas, while CAM 5.2 was most efficient with the small cell carcinomas. CEA stained 33% and 60% of the small and large cell carcinomas, respectively, most adenocarcinomas, and 84% of the squamous cell carcinomas, among which staining decreased with dedifferentiation and was often focal. EMA reacted with 90%, and NSE with 20% of all histological types. There was no staining for NF. All antibodies, except EMA, were more efficient with surgical specimens. Our study implies that the cytokeratins we used work better with surgical material, but are generally comparable to monospecific cytokeratin antibodies. Also, EMA is a reliable marker for epithelial differentiation with all types of tissue samples. Moreover, CEA negativity in several poorly differentiated lung carcinomas might have implications in the differential diagnosis against pleural mesothelioma.

Topics: Carcinoembryonic Antigen; Female; Humans; Immunohistochemistry; Intermediate Filaments; Keratins; Lung Neoplasms; Male; Membrane Glycoproteins; Mucin-1

Morphological analysis of the sclerotic changes in peripheral lung carcinoma (PLC) and nephrosclerosis in renal-cell carcinoma (RCC) established a promoting role of sclerosis in carcinoma development. The pneumosclerosis role as a background process in the PLC development is proved by the following facts: high proportion (83%) of the carcinoma in the scar among PLC; identity of the scar collagen composition in PLC and that in metatuberculosis and metapneumonic pneumosclerosis foci; detection of metatuberculosis foci in 75% of PLC; the presence of the precancerous changes in the epithelium entrapped in the pneumosclerotic foci, not only with signs of morphological atypia, but with the disturbance of nuclear DNA and cellular oncogene expression as well. The association of RCC with nephrosclerosis is shown by a high proportion (82.7%) of the RCC development against the background of nephrosclerosis; the dependence of the so-called cortical adenoma development on the degree of nephrosclerosis; epithelial proliferation in the nephrosclerotic foci with the appearance of undifferentiated cells with the altered DNA content and the expression of cytokeratins and vimentine. Carcinoma morphogenesis against the background of sclerosis may be described as follows: development of sclerosis (focal and/or diffuse), the appearance of the focal epithelial hyperplasia in the scar, dysplasia or adenoma and finally carcinoma.

Topics: Adenoma; Carcinoma, Renal Cell; Cell Differentiation; Cell Division; Collagen; DNA; Humans; Keratins; Kidney Neoplasms; Lung Neoplasms; Nephrosclerosis; Precancerous Conditions; Sclerosis; Vimentin

739 regional lymph nodes from 94 patients with stage I non-small cell lung carcinoma (NSCLC) were studied by immunohistochemistry. These lymph nodes, contained no metastasis as assessed by conventional histopathology, were recut. A series consecutive sections from the original blocks were immunostained with polyclonal and monoclonal antibodies to keratins, carcinoembryonic antigen (CEA) and human milk fat globulin membrane antigen (HMFG-2). Single tumor cells or small clusters of tumor cells, not visible on routine examination, were readily detected. The actual number of lymph nodes that contained occult tumor cells was 123 (16.6%) from 53 patients (56.4%). The majority of 102 immunostaining positive nodes were distributed in the hilar (29%) and peribronchial (25%) regions. Our data indicate that: 1. a series consecutive sections and immunohistochemistry may greatly increase the diagnostic yield of occult micrometastases in lymph nodes. 2. the high incidence of occult metastases in NSCLC may be of importance in relation to their rapid dissemination and high death rate. 3. the high frequency of occult nodal metastases in NSCLC raises questions in regard to our presently used criteria for staging, prognosis and treatment of ostensibly stage I disease. 4. perhaps resections of hilar and peribronchial lymph nodes will have an important clinical significance in prevention of wide dissemination of tumor cells.

Topics: Adult; Aged; Aged, 80 and over; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Membrane Glycoproteins; Middle Aged; Mucin-1

Chemically induced mouse lung tumors exhibit distinctive growth patterns, characterized by an alveolar or solid appearance, a papillary appearance, or a combination of the two. Lung tumors induced in strain A/J mice by either benzo(a)pyrene (BP) or by N-nitrosoethylurea (ENU) were examined for expression of low- and high-molecular-weight cytokeratins. Simple cytokeratins (low molecular weight) were found in all epithelial cells of the normal mouse lung and in all tumor types, whereas higher-molecular-weight cytokeratins were found only in normal bronchiolar cells and in papillary tumor cells. These data lend support to the hypothesis that chemically induced papillary lung tumors in strain A/J mice are derived from bronchiolar Clara cells.

Topics: Adenoma; Animals; Benzopyrenes; Ethylnitrosourea; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Mice; Mice, Inbred A; Microscopy, Electron

Using conventional examination (CE) of H&E stained slides from bone marrow aspirates, metastases can be detected in approximately 25% of patients with small cell lung cancer. We investigated a panel of monoclonal antibodies using immunohistochemistry in the diagnosis of bone marrow infiltration from SCLC and compared the results with CE. Seven monoclonal antibodies raised against epithelial antigens (CAM 5.2, MOV 15, NCCST 433, PE 35, LCA1/L38, HMFG 1 AND HMFG 2) were applied on bone marrow sections from three groups of patients (pts): (1) 19 pts in whom SCLC-metastases were detected by CE, (2) 44 pts with SCLC in whom metastases could not be detected by CE, and (3) 20 pts with non-malignant bone marrow diseases. All the antibodies except LCA1/L38 were positive in 60-90% of the slides with infiltrating tumour cells in group 1. No positive tumour cells were detected in group 2. A few plasma cells and megakaryocytes were slightly positive for MOV 15 and NCCST 433, but no other positive cells were detected in group 3. In conclusion, the monoclonal antibodies used in this study may be useful for diagnostic purposes when a suspicious looking infiltration is detected by CE. However, these antibodies could not detect metastatic tumour cells in the bone marrow sections from patients in whom CE did not reveal any tumour cells.

Topics: Antibodies, Monoclonal; Bone Marrow Diseases; Carcinoma, Small Cell; Humans; Keratins; Lung Neoplasms; Membrane Glycoproteins; Mucin-1; Neoplasm Metastasis; Survival Analysis

The expression patterns of basement membrane components and keratin intermediate filament proteins were studied in normal human bronchial epithelium and 56 lung carcinomas using monoclonal antibodies to laminin, type VII collagen and the individual keratins 14, 16, 17 and 18. In normal lung, laminin and type VII collagen were present between the epithelium and the lamina propria of bronchi and bronchioles. Keratin 14 was expressed in the basal cells, keratin 17 in the basal and some suprabasal cells and keratin 18 in the columnar cells of the bronchi and bronchioles. Keratin 16 was not present in normal bronchial epithelium. Laminin was found in all subtypes of lung carcinoma, but type VII collagen was present only in squamous cell carcinomas, where it showed a reduction in expression with decreasing differentiation. Type VII collagen was not identified in adenocarcinomas, small cell carcinomas or carcinoids. Antibodies to basal cell keratins 14 and 17 also displayed positivity only in squamous cell carcinomas, although no correlation with the degree of differentiation could be observed. Keratin 16 appeared to be a marker of the squamous phenotype, rather than of hyperproliferation. The keratin 18 marker for columnar epithelial cells showed a reaction pattern opposite to that of the basal cell keratins, being extensively present in adenocarcinomas, small cell carcinomas and carcinoids, with less expression in squamous cell carcinomas. This study shows a correlation between the presence of type VII collagen and the basal cell keratins 14 and 17, and a negative correlation between these components and keratin 18. These findings are likely to be useful in identifying lung cancer subtypes.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoid Tumor; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Collagen; Humans; Immunoenzyme Techniques; Keratins; Laminin; Lung Neoplasms

Antibodies to cytokeratins (CKs) have found extensive application in the differential diagnosis of epithelial tumors. The chain-specific anti-CK reagents appear to be of practical value for further subtyping of carcinomas. The authors have produced a novel anti-CK 18 monoclonal antibody (ACK-156) using a modified immunization procedure that included sequential injections of human epidermal keratin, cyclophosphamide, and enriched cytoskeletal extracts from a human lung carcinoma cell line. This protocol effectively amplified clones with reactivity toward CK epitopes not present in epidermal keratin. Monospecificity of the antibody was confirmed by immunoblot analysis using both total cell lysates and cytoskeletal extracts as antigens. Immunoperoxidase staining of adenocarcinomas from a variety of sites, including lung, was strongly positive. Squamous cell carcinomas of lung were also strongly stained whereas squamous cell carcinomas of head and neck origin were stained focally or not at all. In contrast, several commercially available anti-CK 18 monoclonal antibodies did not distinguish squamous cell carcinomas of lung from those of head and neck origin. Immunoblot analysis of tumor lysates corroborated the tissue staining results and revealed that the commercially available antibodies that were tested recognize at least one other low molecular weight peptide in addition to the CK 18 peptide recognized by ACK-156.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoma, Squamous Cell; Cytoskeleton; Electrophoresis, Polyacrylamide Gel; Epitopes; Humans; Hybridomas; Immunoblotting; Immunologic Techniques; Keratins; Lung Neoplasms; Peptide Mapping; Staining and Labeling

Carcinoid tumors of the lung in 10 patients were treated surgically and both the clinicopathological manifestations and immunohistochemistry were examined in detail. Five were central carcinoid tumors, located in the main, lobar or segmental bronchus and five were peripheral carcinoid tumors, located in the subsegmental bronchus or beyond. Histologically, eight of the tumors were typical carcinoid tumors, one was an atypical carcinoid tumor, and one a carcinoid tumorlet. Three growth types were also established: polypoid type, iceberg type and intrapulmonary type. The central carcinoid tumors belonged either to the polypoid type or iceberg type, while the peripheral carcinoid tumors were of the intrapulmonary type. Both the iceberg and intrapulmonary types may invade the peribronchial or parenchymal tissues more frequently than does the polypoid type. Immunohistochemically, argyrophilia and neuron-specific enolase (NSE) were detected in all the tumors examined and six stained for polypeptide hormones such as adrenocorticotropic hormone (ACTH) and/or pancreatic polypeptide (PP). Of these, five had epithelial markers such as keratin, epithelial membrane antigen (EMA) and/or carcino-embryonic antigen (CEA). These findings suggest that a carcinoid tumor of the lung originates from primitive multipotential stem cells such as those of a neuroendocrine or epithelial nature.

Topics: Adrenocorticotropic Hormone; Adult; Aged; Antigens, Neoplasm; Carcinoembryonic Antigen; Carcinoid Tumor; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; Pancreatic Polypeptide; Phosphopyruvate Hydratase

At clinical presentation, the majority of malignant tumors are composed of multiple clonal subpopulations of tumor cells with different phenotypic characteristics. Using the experimental tumor model ER 15-P, a methylcholanthrene-induced pleomorphic sarcoma of the C57 Bl6J mouse, we studied a system of long-term in vivo passages of this primary tumor for cell morphological changes, and alterations in the potential for spontaneous lung metastases. Transplants from the primary after the 4th, 20th, 40th and 80th i.m. passage (referred to as T4, T20, T40, and T80 respectively) together with their lung metastases were investigated by light microscopy, immunohistochemistry, and electron microscopy. In addition, the potential for metastasis to the lungs in each group was determined and compared with that of the parent T4 tumors. T4 tumors were mainly composed of spindle-shaped tumor cells with the ultrastructural features of fibroblasts and myofibroblasts, often arranged in a storiform or fasciculated growth pattern, and intermingled with tumor giant cells. Some small areas contained polygonal or rounded tumor cells, ultrastructurally undifferentiated, and sometimes arranged in a hemangiopericytoma-like growth pattern. Although electron-microscopical findings clearly demonstrated the mesenchymal origin of these tumor cells, immunostaining with a polyclonal antibody to vimentin was unspecific in all tumor cells and normal mouse tissue. Monoclonal antibodies to vimentin from different sources were completely negative in tumor cells and murine stromal components. In contrast, myofibroblast-like tumor cells showed immunohistochemically, a moderate to strong co-expression with monoclonal antibodies to desmin, muscle actin and alpha-smooth muscle actin. On the basis of these morphological findings, the primary ER 15-P was classified as a pleomorphic myofibrosarcoma. The lung metastases of T4 tumors were mainly composed of undifferentiated round to polygonal tumor cells, while the number of desmin-positive, muscle- and alpha-smooth muscle-actin-positive cells was reduced. The morphological features of T20 tumors and their lung metastases were the same as in T4, indicating a relative stability of the phenotype up to that stage. In contrast, T40 and T80 tumors and their lung metastases were found to contain almost exclusively undifferentiated tumor cells and many tumor giant cells. While fibroblast-like tumor cells were seen only occasionally, myofibroblast-like tumor cel

Topics: Actins; Animals; Disease Models, Animal; Female; Fibrosarcoma; Keratins; Lung Neoplasms; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Phenotype; Sarcoma, Experimental

This paper is a report of a large cell carcinoma (solid carcinoma with mucus formation-WHO) which developed in the right upper lobe of a 71-year-old woman and in which cytoplasmic Mallory body-like inclusions (MBL) similar to Mallory bodies (MB) often found in alcoholic liver disease and hepatocellular carcinoma were noted in the tumor cells. MBL was an eosinophilic massive or reticular hyalin, and showed staining properties similar to those of MB in general stainings. Electron microscopically, it was made up mainly of a fine granular amorphous substance with high electron density and was not surrounded by a membrane. A part of the margin was in close contact with filaments approximately 10 nm in diameter, with tonofibril-like structures around it, which suggested a relationship with cytokeratin. On immunoperoxidase staining using a couple of cytokeratin antibodies, definitely positive findings were not obtained with MBL themselves. MBL of this case basically has the same character as that of MBL which appears in fibrous lesions in the lung and MB in liver disease. It was concluded that it is necessary to preoperatively distinguish this entity cytologically or histologically from pulmonary metastasis of hepatocellular carcinoma.

Topics: Aged; Carcinoma, Hepatocellular; Carcinoma, Small Cell; Diagnosis, Differential; Female; Humans; Inclusion Bodies; Keratins; Liver Neoplasms; Lung Neoplasms

On review of 115 poorly or undifferentiated lung cancers from 671 lung tumors resected over a 7-year period, we have found 38 cases of basaloid carcinoma. The cardinal histopathologic features distinguishing this tumor from other non-small cell lung cancers are a lobular growth pattern of small cells with moderately hyperchromatic nuclei, with no prominent nucleoli, and with scant cytoplasm, a high mitotic rate, and peripheral palisading. Basaloid carcinoma was present in a pure form in 19 cases and the other 19 tumors were of a mixed, but prominent, basaloid type associated with squamous cell carcinoma, large cell carcinoma, or adenocarcinoma. The immunophenotype of basaloid cancers was close to that of basal bronchial epithelial cells, with a low level of expression of low molecular weight cytokeratins. Staining for neuroendocrine markers was infrequent and inconsistent. Ultrastructural study showed an absence of neurosecretory granules and the presence of some squamous and/or glandular differentiation. This morphologic and immunologic phenotype suggests that basaloid carcinoma is derived from a pluripotent reserve cell or a basal bronchial epithelial stem cell. This unique histologic form of lung tumor has a poor prognosis, with a median survival rate of 22 months for stage I and II disease. This justifies classification of basaloid carcinoma as a distinct form of lung cancer, separate from small cell lung carcinoma.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Basal Cell; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Microscopy, Electron; Middle Aged; Neoplasm Staging; Phenotype; Prognosis; Survival Analysis

Three human lung squamous carcinoma cell lines (NX002, CX140 and CX143) demonstrate features of squamous differentiation including involucrin synthesis and competence to form cornified envelopes. 12-O-Tetradecanoylphorbol 13-acetate inhibits growth of these cell lines and this growth inhibition is associated with enhanced differentiation.

Topics: Antigens, Neoplasm; Carcinoma, Squamous Cell; Cell Differentiation; Cell Division; Humans; Keratins; Lung Neoplasms; Protein Precursors; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

Adenomyoepitheliomas of the breast have been considered to have limited metastatic potential; axillary node metastasis has been reported, but there has been no report of distant metastasis. We report six cases, including two malignant adenomyoepitheliomas, one of which metastasized to the lung and brain. Patient age ranged from 26 to 63 years (mean 46). Primary tumors were solitary and measured 0.9-3.5 cm (mean 1.7). Five of six tumors presented as palpable masses. Two patients treated by local resection have no evidence of disease at 5 and 18 months' follow-up. Two patients treated by local resection had recurrences, one at 48 the other at 60 months. The fifth patient had a spindle-cell type adenomyoepithelioma diagnosed as malignant because of high mitotic rate and cytologic atypicality of the myoepithelial component. This patient was treated by mastectomy and has no evidence of disease at 18 months. The sixth patient, initially treated by local excision, had six local recurrences over 52 months treated by reexcisions, mastectomies, and radiation. A lung metastasis was resected at 54 months and brain metastases were identified at 60 months with death occurring at 64 months. Both malignant adenomyoepitheliomas had high mitotic rates [11-14/10 high-power fields (HPF)] diffusely throughout the tumors and foci of cytologically malignant cells. The malignant adenomyoepithelioma that metastasized had an infiltrative growth pattern that increased with successive local recurrences. The four other tumors had only isolated areas of mitotic activity (maximum 1-9/10 HPF) and minimal cytologic atypia. Immunohistochemistry performed on five of six cases confirmed dual epithelial/myoepithelial cell populations in all tumors examined, including the metastasis. Electron microscopic examination of the malignant adenomyoepithelioma that metastasized also confirmed dual epithelial/myoepithelial cell populations in a local recurrence and the lung metastasis. We conclude that there is a spectrum of behavior for breast adenomyoepitheliomas with potential for local recurrence and, rarely, distant metastasis.

Topics: Adult; Brain Neoplasms; Breast Neoplasms; Carcinoembryonic Antigen; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Membrane Glycoproteins; Microscopy, Electron; Middle Aged; Mitotic Index; Mucin-1; Myoepithelioma; Neoplasm Metastasis; Recurrence

Despite the high response rates resulting from chemotherapy, the majority of small-cell lung cancer (SCLC) patients relapse with chemoresistant tumors. To analyze the phenotypic changes that are precursors of chemoresistant status, and to investigate the role of chemotherapy in these changes, tumor samples from 20 patients, taken before chemotherapy (etoposide, doxorubicin, and cyclophosphamide) and again at the onset of chemoresistance (after at least three courses of chemotherapy), were compared. The histologic changes were minor in 10 of 20 patients, as shown by an increase in cell size; they were major in 10 of 20 patients, with the appearance of mixed composite tumors in which neuroendocrine (NE), epidermoid, and glandular components were mixed. Major changes correlated with a good response to chemotherapy (P = .001). Ultrastructural studies showed an increase in neurosecretory granules and desmosomes, and a high frequency of multidirectional differentiation (45%) when comparison was made with pretherapy samples (10%) (P less than .01). Immunohistochemical (IH) analysis showed an increase in cytokeratin (CK) expression in treated patients, with a different labeling pattern and the expression of higher molecular weight CK. The expression of NE lineage markers (Leu 19, Sy 38, SL 11-14) remained stable, while that of NE differentiation markers (Leu 7, chromogranin) increased in the treated patients. The neuron-specific enolase (NSE) activity remained stable in treated SCLC. Large cells with a more differentiated phenotype and proliferative capacity (as shown by Ki 67 labeling), appeared to be characteristic of treated and secondary chemoresistant SCLC. The acquisition of a more complex phenotype, which correlates with primary response to therapy, implies a drug-induced differentiation in SCLC.

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Small Cell; Cell Differentiation; Cyclophosphamide; Cytoplasmic Granules; Desmosomes; Doxorubicin; Drug Resistance; Etoposide; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Molecular Weight; Neoplasm Staging; Phenotype; Phosphopyruvate Hydratase; Survival Rate

A panel of seven monoclonal antibodies including anti-vimentin, anti-keratin markers AE1/AE3 and EAB902, human milk fat globule (HMFG-2), B72.3, anti-carcinoembryonic antigen (CEA), and anti-Leu-M1 were used for an immunoperoxidase staining assay to determine their value in the differentiation of pleural mesothelioma from lung adenocarcinoma. Anti-vimentin positively identified 86% of the mesotheliomas and none of the adenocarcinomas. AE1/AE3, EAB902, and B72.3 reacted with a high percentage of both mesothelioma and adenocarcinoma specimens. With HMFG-2, both membrane and cytoplasmic staining was observed in 92% of the adenocarcinomas and in 14% of the mesotheliomas, whereas 26% of the mesotheliomas only exhibited membrane staining. Eighty percent of the adenocarcinomas and 8% of the mesothelioma tissues stained with anti-Leu-M1. Anti-CEA did not react with any of the 50 mesotheliomas tested but did react with 95% of the lung adenocarcinomas tested. From this study, it was concluded that anti-CEA and anti-Leu-M1 were the most effective of the seven tumor markers evaluated; and that 100% of the pleural mesothelioma tissues could be correctly differentiated from lung adenocarcinomas using a panel consisting of anti-vimentin, HMFG-2, anti-CEA and anti-Leu-M1 monoclonal antibodies.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Differentiation, Myelomonocytic; Antigens, Neoplasm; Carcinoembryonic Antigen; Diagnosis, Differential; Glycoproteins; Humans; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Lung Neoplasms; Membrane Glycoproteins; Mesothelioma; Mucin-1; Pleural Neoplasms; Vimentin

A 19-month-old black girl had a radical nephrectomy for a Wilms' tumor that contained areas of epithelium indistinguishable from renal cell carcinoma. She was treated with chemotherapy but subsequently had pulmonary metastases develop and massive abdominal recurrence. The recurrent tumor was histologically renal cell carcinoma with no identifiable Wilms' tumor elements. The child died with recurrent and metastatic tumor 13 months after nephrectomy. Pathologic, immunoperoxidase, and flow cytometric studies of this unusual case are presented.

Topics: Carcinoma, Renal Cell; Female; Humans; Immunoenzyme Techniques; Infant; Keratins; Kidney Neoplasms; Lung Neoplasms; Ploidies; Recurrence; Vimentin; Wilms Tumor

Monoclonal anticarcinoembryonic antigen (antiCEA), human milk factor globulin (HMFG2), and antikeratin antibodies were assessed for their value in the differential diagnosis of pleural mesothelioma (53 cases) and carcinoma of the lung (60 cases) in material from necropsies. In 40 of the cases pleural biopsies were also studied in the same manner. AntiCEA was found to be the best discriminating antibody for most types of mesothelioma; HMFG2 was slightly less valuable but a useful additional tool. Antikeratin was the least useful. For both antiCEA and HMFG2 antibodies, however, the proportion of carcinomas staining was smaller than in previous studies and this, combined with the positive staining of some mesotheliomas, reduces the value of the reactions in the individual case. Medical panels adjudicating compensation claims should not use these reactions as the sole criteria in deciding the origin of the tumours in these cases.

Topics: Antigens, Neoplasm; Autoantibodies; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma; Diagnosis, Differential; Humans; Keratins; Lung Neoplasms; Membrane Glycoproteins; Mesothelioma; Mucin-1; Occupational Diseases; Pleural Neoplasms

A case of disseminated pulmonary carcinoma is presented. Metastases of both carcinoma and sarcoma were confirmed via light microscopy and immunohistological examination. Our observations on this rare tumour are compared with a review of the literature.

Topics: Carcinosarcoma; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Neoplasm Metastasis; Vimentin

Two cases of monophasic spindle cell carcinomas and one case of adenosquamous carcinoma with the spindle cell component located in the lower respiratory tract are presented. In the biphasic tumor, areas of transition from carcinoma to sarcomatous spindle cells were clearly found. The two monophasic tumors and the spindle cell component of the biphasic tumor were histologically characterized by sheets of spindle cells. However, by electron microscopic and immunohistochemical study, several features of squamous epithelial differentiation were found in the spindle cell areas of all cases. Keratin and vimentin were, in various degrees, coexpressed in all the cases. Therefore, it is supposed that the spindle cell component displays a spectrum of phenotypes originating from squamous cell carcinoma, and monophasic spindle cell carcinoma is considered as a kind of the extreme phenotype of squamous cell carcinoma pretending mesenchymal differentiation.

Topics: Adenocarcinoma; Adult; Carcinoma; Carcinoma, Squamous Cell; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Vimentin

Topics: Aged; Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; Biopsy, Needle; Bone Marrow; Carcinoma, Bronchogenic; Carcinoma, Small Cell; Epithelium; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Membrane Glycoproteins; Mucin-1; Phosphopyruvate Hydratase

Cytogenetic analysis of two pulmonary chondroid hamartomas and nine breast adenofibromas revealed clonal chromosome aberrations in both hamartomas and in four breast tumors. To determine lineage of the cells with chromosome aberrations, a combined immunohistochemical/cytogenetic approach was developed that enabled simultaneous ascertainment of cytogenetic aberrations and immunohistochemical features in individual cells. Immunohistochemical/cytogenetic evaluation of one hamartoma and two adenofibromas demonstrated that neoplastic proliferation, in each case, was confined to the mesenchymal (stromal) component, whereas epithelial cells appeared to be reactive. Cytogenetically abnormal short-term cultures of the remaining hamartoma and another of the breast adenofibromas were composed entirely of mesenchymal elements, indicating mesenchymal clonality in those tumors as well. Our findings support redesignation of pulmonary chondroid hamartomas as 'pulmonary chondromas' and suggest that carcinomas developing within fibroadenomas arise from reactive epithelial proliferation. Combined immunohistochemical/cytogenetic analysis might be useful in the development of novel therapeutic approaches that selectively target neoplastic populations within solid tumors.

Topics: Adenocarcinoma; Breast Neoplasms; Cell Transformation, Neoplastic; Chromosome Aberrations; Hamartoma; Humans; Immunohistochemistry; Karyotyping; Keratins; Lung Neoplasms; Mesoderm; Trisomy; Tumor Cells, Cultured; Vimentin

A preoperative diagnosis of pulmonary blastoma was established by fine-needle aspiration (FNA) cytology in a patient who presented with a pulmonary mass discovered incidentally on chest x-ray. At the time of computed tomography-guided FNA, an on-site preliminary diagnosis of poorly differentiated carcinoma was made after examination of a rapid hematoxylineosin (H&E)-stained smear. Malignant epithelial and stromal elements could be demonstrated on smears subsequently stained with the H&E, Papanicolaou, and Diff-Quik methods. The cell block preparation showed a distinctly biphasic malignant tumor with classic morphologic features of pulmonary blastoma. We present the clinical, cytologic, immunocytochemical, and histologic findings of this case and re-emphasize the valuable contribution of adequate cell block preparations to accurate diagnosis of fine-needle aspiration material.

Topics: Actins; Biopsy, Needle; Cytodiagnosis; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Staining and Labeling; Vimentin

Based on our review of 35 cases and the literature, we found the spectrum of pulmonary neuroendocrine (NE) tumors to be too broad to fit into the traditional three-category classification scheme of typical carcinoid (TC), atypical carcinoid (AC), and small-cell lung carcinoma (SCLC). We found that a spectrum of high- and low-grade tumors exist between TC and SCLC and that in the past many of these tumors have been called AC. We chose to adhere to Arrigoni's definition of AC, as his original criteria characterized a low-grade tumor. For the higher grade non-small-cell tumors (NSCLC), we propose a fourth category of large-cell neuroendocrine carcinoma (LCNEC), which is characterized by: (a) light microscopic NE appearance; (b) cells of large size, polygonal shape, low nuclear-cytoplasmic ratio (N:C), coarse nuclear chromatin, and frequent nucleoli; (c) high mitotic rate [greater than 10/10 high-power fields (HPF)] and frequent necrosis; and (d) NE features by immunohistochemistry (IHC) or electron microscopy (EM). Thus, after deciding that a pulmonary NE tumor is high grade, the major diagnostic issue is separation of LCNEC from SCLC. This distinction is based not only on cell size, but on a variety of morphologic features. We studied 20 TC, six AC, five LCNEC, and four SCLC and characterized the clinical, light microscopic, EM, IHC, and flow cytometric features of each type of tumor. We did not find any advantage to IHC, EM, or flow cytometry over light microscopy in the subclassification or prediction of prognosis; however, these methods were useful in characterizing these four types of pulmonary NE tumors and in demonstrating their NE properties. LCNEC must be distinguished from a fifth category pulmonary NE tumor: NSCLC with NE features in which NE differentiation is not evident by light microscopy and must be demonstrated by EM or IHC. Although the prognosis of LCNEC appears to be intermediate between AC and SCLC, larger numbers of patients will be needed to demonstrate significant differences in survival.

Topics: Adrenocorticotropic Hormone; Adult; Aged; Antigens, Differentiation; Bombesin; Calcitonin; Carcinoembryonic Antigen; Carcinoid Tumor; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; CD57 Antigens; Chromogranins; Female; Flow Cytometry; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Membrane Proteins; Microscopy, Electron; Middle Aged; Synaptophysin; Terminology as Topic

Two intraparenchymal lung tumors exhibiting the histopathologic and immunophenotypic characteristics of an intracranial meningioma are presented. The meningiomas presented as solitary asymptomatic nodules in elderly individuals. Both patients survived longer than 3 years following resection, and neither displayed clinical or radiographic evidence of a central nervous system tumor, suggesting that these are primary lung tumors. Review of the literature and discussion of other lesions in the differential diagnosis of this rare intrapulmonary neoplasm are presented.

Topics: Aged; Female; Humans; Immunoenzyme Techniques; Immunophenotyping; Keratins; Lung Neoplasms; Membrane Glycoproteins; Meningioma; Middle Aged; Mucin-1; S100 Proteins; Vimentin

Three commonly used keratin monoclonal antibodies (MAB)--AE1:AE3, CAM 5.2, and MAK-6--were compared with routinely used cytokeratin antibody. The expression of these antibodies was analyzed in several tissues obtained from clinically normal dogs and in a variety of neoplasms from dogs. Using appropriate enzymatic digestion, paraffin-embedded tissues processed in routine manner retained their typical keratin expression. Differentiated and poorly differentiated epithelial neoplasms, lymphomas, and melanomas were studied by use of the avidin-biotin-peroxidase technique. All 4 of the aforementioned antibodies had similar staining profiles. Of 3 anaplastic carcinomas, 2 had positive reaction to all 4 antibodies. All lymphomas, plasma cell tumors, and amelanotic melanomas had negative reaction to MAK-6, CAM 5.2, AE1:AE3, and cytokeratin MAB. Three basal cell epitheliomas had positive reaction to all 4 antibodies, whereas 1 basal cell tumor with a solid pattern had negative staining reaction. Two carcinoids had negative reaction to all markers and 1 of 2 malignant chemodectomas and 1 transitional cell carcinoma had staining reaction to only AE1:AE3 MAB. Comparing the 4 antibodies, use of AE1:AE3 MAB produced the strongest staining intensity followed by cytokeratin, MAK-6, and CAM 5.2 MAB. All 4 antibodies had low background staining. In conclusion, AE1:AE3 and MAK-6 MAB are as useful as cytokeratin MAB for identification of poorly differentiated epithelial neoplasms in dogs and cats.

Topics: Animals; Antibodies, Monoclonal; Carcinoma; Dog Diseases; Dogs; Immunohistochemistry; Intestine, Small; Keratins; Lung Neoplasms; Neoplasms; Predictive Value of Tests; Urinary Bladder

We have previously described the neoplastic transformation of immortalized human bronchial epithelial cells (BEAS-2B) by the combination of the c-raf-1 and c-myc protooncogenes and the concomitant induction of neuron-specific enolase mRNA expression (A. Pfeifer et al., Proc. Natl. Acad. Sci. USA, 86: 10075-10079, 1989). In this paper we describe the morphological, biochemical, and immunohistochemical characteristics of the primary c-raf-1/c-myc tumors, xenografts of these tumors, and tumors that originated from cell lines of the primary neoplasm. The tumors were morphologically characterized by the appearance of desmosomes and tonofilaments, microvilli, and dense core granules representing markers of squamous, glandular, and neuroendocrine differentiation, respectively. A total of 11 of 13 tumors were positive by immunohistochemical techniques for neuron-specific enolase, serotonin (nine of 13), and calcitonin (six of 13). Keratins were expressed in 11 of 13 tumors, and while specific keratins (K5, K7, K16/K17) decreased, there was an increase of vimentin in the tumor cells. Gastrin-releasing peptide immunoreactivity was detectable in a small number of tumors (five of 13). BEAS-2B cells transfected with the c-raf-1 and c-myc protooncogenes and cell lines established from the primary tumors expressed major histocompatibility Class II antigen which has been found on small cell lung carcinoma cells. The tumors induced by the c-raf-1 and c-myc protooncogenes resemble the multidifferentiated phenotype of small cell lung cancer frequently detected in vivo and present a defined model to study the relation between molecular markers, phenotypical appearance, and response to chemotherapeutic agents and radiation.

Topics: Adenocarcinoma; Animals; Antigens, Neoplasm; Bronchi; Carcinoma, Squamous Cell; Cell Line; Cell Transformation, Neoplastic; Epithelium; Genes, myc; Histocompatibility Antigens Class II; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mice; Neoplasm Transplantation; Phosphopyruvate Hydratase; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-raf; Proto-Oncogenes; Transplantation, Heterologous

Atypical mesothelial hyperplasia encountered in pleural fluid or in a pleural biopsy specimen raises the suspicion that one may be dealing with a diffuse malignant mesothelioma of the pleura. We studied eight cases with cytologic or histologic changes of mesothelial atypia thought to be suspicious for diffuse malignant mesothelioma. In each case, the hyperplasia was associated with a bronchogenic carcinoma in the lung subjacent to the mesothelial hyperplasia. Bronchogenic carcinoma should be added to the list of causes of atypical mesothelial hyperplasia. This combination of reactive and malignant processes should be appreciated, since pleural carcinomatosis and diffuse malignant mesothelioma must be separated for clinical and epidemiologic reasons.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Carcinoma, Bronchogenic; Carcinoma, Squamous Cell; Female; Humans; Hyperplasia; Keratins; Lung; Lung Neoplasms; Male; Middle Aged; Pleura; Pleural Effusion, Malignant

This paper presents an immunocytochemical study performed on cytocentrifuged deposits from 109 peritoneal and pleural effusions including 20 transudates, 43 malignant metastatic effusions and 46 effusions containing atypical cells, unidentifiable as reactive mesothelial or malignant epithelial cells on the classical morphological criteria. A panel of four monoclonal antibodies (MAb) was used, including KL1 directed to cytokeratins (KER), V9 to vimentin (VIM), NEO 723 to carcinoembryonic antigen (CEA) and E29 to epithelial membrane antigen (EMA). In most transudates the reactive mesothelial cells coexpressed VIM and KER with a ring-like pattern for the latter proteins. In contrast, they were unreactive to anti-CEA and weakly and inconsistently reactive to anti-EMA. In malignant effusions, most carcinoma cells coexpressed EMA, CEA and KER with a predominant diffuse cytoplasmic pattern for the latter. Only a few malignant epithelial cells from five metastatic adenocarcinomas weakly expressed VIM. When used on the 46 effusions with unidentifiable cells, the panel of MAb allowed reactive mesothelial cells and malignant epithelial cells to be distinguished from each other in 39 of 46 cases (85%).

Topics: Adenocarcinoma; Antibodies, Monoclonal; Ascitic Fluid; Carcinoembryonic Antigen; Carcinoma; Cytodiagnosis; Epithelium; Exudates and Transudates; Humans; Immunohistochemistry; Immunophenotyping; Keratins; Lung Neoplasms; Membrane Glycoproteins; Mucin-1; Pleural Effusion, Malignant; Vimentin

We used immunohistochemical procedures to study the cellular expression and distribution of cytokeratins (CKs) in rat 13762NF mammary adenocarcinoma cells growing at mammary fat pad sites and at spontaneous lymph node and lung sites. In order to establish CK distribution in normal rat mammary epithelia, immature, resting, and lactating rat mammary glands were probed with a panel of monospecific antibodies that recognize individual CKs. Basal/myepithelial cells were distinguished by expression of CKs 5 and 14 and coexpression of vimentin from luminal cells, which expressed CKs 8, 18, and 19. Antibody to CK 7 recognized luminal epithelium of immature and resting, but not lactating, mammary glands. Myoepithelial cells of lactating mammary gland were weakly recognized by antibodies to CKs 7 and 19. Tumors formed by cell lines and clones derived from parental 13762NF tumor (MTPa, MTC, MTA, and MTF7) were not recognized by any of the anti-CK antibodies. Only vimentin was expressed in these tumors and their metastases. In tumors and metastases generated from cell lines and clones derived from lymph node (MTLY) and lung metastases (MTLn2 and MTLn3) of the 13762NF tumor we observed heterogeneous CK phenotypes. Expression of CKs 5 and 18 was greatly reduced or lacking, while CK 14 was coexpressed with CKs 7, 8, and 19 with or without vimentin. Tumors from the highly metastatic clone MTLn3 had a dominant cellular phenotype, expressing CKs 7, 8, 14, and 19 and vimentin, a pattern that did not match normal mammary epithelia, whether luminal, basal/myoepithelial, or the dual-phenotype stem cell, in which CKs 5, 8, 14, and 18 were coexpressed. MTLn3 lymph node and lung metastases expressed the same cellular phenotype as the s.c. growing MTLn3 tumor. The results appear to contradict the belief that malignant mammary tumors may be distinguished from benign tumors or hyperplastic growths by the lack of basal/myoepithelial markers.

Topics: Adenocarcinoma; Animals; Animals, Newborn; Antibodies, Monoclonal; Biomarkers, Tumor; Cell Line; Female; Fluorescent Antibody Technique; Gene Expression; Keratins; Lung Neoplasms; Lymphatic Metastasis; Mammary Glands, Animal; Mammary Neoplasms, Experimental; Neoplasm Metastasis; Phenotype; Rats; Vimentin

The development of antibodies to DNA-incorporated thymidine analogs has in turn led to the development of flow cytometric techniques for rapidly measuring cell kinetics parameters. More recently, these techniques have been applied to clinical tumor material. One problem with such measurements has been the difficulty of distinguishing malignant cells from coexistent normal cells in the biopsy material. In the present study, the feasibility of selecting out the desired malignant cell population for kinetic analysis from a mixture of cells was tested in vitro. An anticytokeratin antibody was used to discriminate between a mixture of tumor cells (cytokeratin positive) and normal cells (cytokeratin negative). The cell lines chosen for the study, A549 human lung carcinoma cells and Chinese hamster ovary (CHO) cells, were pulse labeled with iododeoxyuridine (IdUrd) and sampled every hour up to 16 hours. Selecting out cells from the mixture required the application of three-color fluorescence flow cytometry, which was carried out using the fluorochromes FITC (fluorescein isothionate, green fluorescence, IdUrd-DNA antibody), PE (phycoerythrin, orange fluorescence, cytokeratin antibody), and PI (propidium iodide, red fluorescence, DNA). This allowed single laser excitation. The staining procedure involved incubation with the IdUrd antibodies (specific antibody plus FITC-conjugated second antibody) followed by the cytokeratin antibodies (specific antibody plus PE-conjugated second antibody) and lastly by the DNA stain containing RNase. Two analysis methods of the IdUrd/DNA cytograms were applied: a mid-S window analysis and a relative movement (RM) analysis. Results of the analyses for cells selected out of mixtures were compared with results of cells stained and analyzed separately. A clear separation of the two cell lines could be obtained on the basis of orange fluorescence (cytokeratin content) despite a large overlap of their DNA histograms. By gating on high or low orange fluorescence, almost pure populations of the individual cell types could be selected out for further kinetic analysis. Little difference was seen, with both the mid-S and RM analyses, between cells gated from mixtures or stained separately. It is concluded that this technique is feasible for use on clinical material, provided good cell suspensions can be obtained, leading to the hope of increasing the accuracy of kinetic measurements on human tumors.

Topics: Animals; Antibodies; Cell Cycle; Cell Line; Cricetinae; Cricetulus; DNA; DNA, Neoplasm; Female; Fibroblasts; Flow Cytometry; Fluorescence; Humans; Keratins; Lung Neoplasms; Ovary; Tumor Cells, Cultured

Extravascular coagulation and fibrinolysis are intimately involved in and modulate cancer cell growth, invasion and metastasis. Samples from resection specimens of patients with primary lung cancer (adenocarcinomas) were tested with monoclonal (MAb) and polyclonal (PAb) antibodies against various factors of the coagulation or fibrinolysis systems, or against antigens of inflammatory or proliferating cells. MAb Ki-67 specific to nuclear antigens of proliferating cells showed a distinct but variable staining of cell nuclei throughout the tumor tissue. Nests of tumor tissue stained with cytokeratin-specific antibodies (PKK1), whereas other parts were negative. Fibrin(ogen) and fibronectin were found throughout the tumor tissue stroma and in the alveolar lining, and the most densely stained areas were at the transition zone between normal and tumor tissue. Fibrinolytic system components like tissue plasminogen activators (t-PA), and urokinase (u-PA), and their inhibitors PAI-1 and PAI-2 were all studied. All specimens were negative for t-PA (except endothelial linings), whereas urokinase-specific antibodies stained loosely packed tumor cells and macrophages within the tumor stromal tissue and alveolar septa. Both PAI-1 and PAI-2 were most prominently expressed within interstitial and alveolar macrophages. A weaker staining of tumor tissue cells was demonstrated. Inflammatory cells like macrophages and T lymphocytes were located in aggregates or diffusely spread within tumor stromal tissue. The inflammatory reaction was most intense at the border between normal lung and tumor tissue.

Topics: Adenocarcinoma; Antibodies; Antibodies, Monoclonal; Biomarkers; Blood Coagulation; Fibrin; Fibrinogen; Fibrinolysis; Fibronectins; Humans; Immunohistochemistry; Inflammation; Keratins; Ki-67 Antigen; Lung Neoplasms; Macrophages; Nuclear Proteins; Plasminogen Inactivators; T-Lymphocytes; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

By indirect immunofluorescence microscopy and electron microscopy, we studied the behavior of intermediate filaments during mitosis in three human epithelial cell lines, derived from normal epidermis (PcaSE-1, from a cancer patient), stratified epithelium (CNE, from nasopharyngeal carcinoma) and simple epithelium (SPC-A-1 from lung adenocarcinoma) respectively. CNE cells and SPC-A-1 cells express two different intermediate filament systems; keratin filaments and vimentin filaments, but PcaSE-1 cells only express keratin filaments. The keratin filament system in PcaSE-1 cells remained intact and encircled the developing mitotic spindle as the cells entered mitosis. In contrast, in CNE cells and SPC-A-1 cells, keratin filaments appeared to disassemble into amorphous cytoplasmic bodies during mitosis. However, their vimentin filaments remained morphologically intact throughout mitosis. We propose; (1) The disassembly of keratin filaments in mitotic epithelial cells is more or less associated with the degree of their cell malignancy rather than with the abundance of keratin filaments in interphase. (2) Intermediate filaments may be involved in the positioning and/or centering of the spindle during mitosis. (3) The possible function of vimentin filament system in CNE cells is positioning and orientation of chromosomes.

Topics: Adenocarcinoma; Cell Line; Epithelial Cells; Epithelium; Humans; Intermediate Filaments; Keratins; Lung Neoplasms; Mitosis; Nasopharyngeal Neoplasms; Tumor Cells, Cultured; Vimentin

Topics: Breast Neoplasms; Female; Histiocytoma, Benign Fibrous; Humans; Keratins; Lung Neoplasms; Middle Aged; Phyllodes Tumor

We studied an unusual sarcoma with morphologic features diagnostic of epithelioid sarcoma by conventional light microscopy, transmission electron microscopy, and immunohistochemistry. The primary tumor, which was located in the deep soft tissues of the buttock of a 32-year-old woman, and its metastases to lymph nodes, liver, and lung were available for investigation. The histomorphological and ultrastructural appearance of the primary tumor and its metastatic deposits were typical of epithelioid sarcoma. Immunohistochemistry revealed a strong and uniform reactivity for vimentin in both the primary tumor and its metastases. In contrast, a marked cytoskeletal heterogeneity became evident for cytokeratins and neurofilaments, which were observed exclusively in lymph node metastasis. To our knowledge, the observation of neurofilaments in epithelioid sarcoma has not previously been reported.

Topics: Adult; Buttocks; Cytoskeleton; Endoplasmic Reticulum; Female; Humans; Immunohistochemistry; Intermediate Filaments; Keratins; Liver Neoplasms; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Microscopy, Electron; Sarcoma; Vimentin

Monoclonal antibodies (MAbs) to specific keratin subtypes were prepared and characterized by immunoblotting and immunohistochemical assays on human cell cultures and normal and malignant human tissues. Chain-specific MAbs to keratin 7 (RCK 105, OV-TL 12/30) and keratin 18 (RGE 53, RCK 106, CK18-2), as well as broadly cross-reacting keratin MAbs (RCK 102, OV-TL 12/5) could be shown to react with different types of human epithelial tissues and were therefore tested for their usefulness in the differential diagnosis of carcinomas. The two broad-spectrum antibodies stained virtually all of the more than 350 carcinomas tested, especially when combined, and distinguished them from most nonepithelial tumors. The keratin 18 MAbs distinguished adenocarcinomas (which are keratin 18 positive) from most squamous cell carcinomas (which are generally keratin 18 negative). The MAbs to keratin 7 could be shown to recognize specific subtypes of adenocarcinoma and could, for example, distinguish between ovarian carcinomas (keratin 7 positive) and carcinomas of the gastrointestinal tract (keratin 7 negative), or between transitional cell carcinomas (keratin 7 positive) and prostate cancer (keratin 7 negative). In general, malignancies showed the expected keratin reactivity pattern as concluded from the keratin pattern of its cell of origin or its type of differentiation. The use of an extended series of malignancies did, however, also illustrate that exceptions to this rule exist. For example, certain antibodies to keratin 18 stained tumor areas in squamous cell carcinomas of the lung. Also a certain percentage of tumors, which generally showed no keratin 7 expression, were positive with RCK 105 or OV-TL 12/30. On the other hand, a certain percentage of tumors, which were generally positive for keratin 7, did not show a staining reaction with these MAbs. Furthermore subtle differences between reactivity patterns of different MAbs recognizing the same keratin protein were observed, both in the normal and malignant human tissues, indicating that specific keratin epitopes may be masked in certain tissues and that unmasking of such epitopes can occur with malignant progression. This phenomenon may be of some use in a further subtyping of carcinomas, especially those of the gastrointestinal tract. Despite these exceptional staining patterns, the keratin MAbs described above have proved to be useful tools in the characterization of epithelial tumors in routine histopathology and

Topics: Adenocarcinoma; Adult; Aged; Antibodies, Monoclonal; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Gastrointestinal Neoplasms; Histocytochemistry; Humans; Immunoblotting; Keratins; Lung Neoplasms; Male; Middle Aged; Ovarian Neoplasms; Prostatic Neoplasms

The relationship between the nuclear DNA content, the immunohistochemical findings, the clinical characteristics (tumor volume doubling time and survival) and the cytomorphologic features of small cell poorly differentiated squamous cell carcinoma of the lung was studied in ten cases. There were no significant correlations between the immunohistochemical stainings for neuron-specific enolase and keratin and the clinical characteristics in these cases. The DNA histogram patterns were classified as type I or type II, depending on the degree of dispersion of values. There was no relationship between the immunohistochemical findings and the DNA histogram patterns. Only the DNA histogram patterns were related to some of the clinical characteristics: patients with type II histograms had significantly shorter tumor volume doubling times than did patients with type I histograms. Such information may aid in distinguishing the small cell type of poorly differentiated squamous carcinoma from classic small cell carcinoma of the lung, with which it may be confused.

Topics: Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; DNA, Neoplasm; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Prognosis

Topics: Adult; Aged; Antibodies, Monoclonal; Breast Neoplasms; Cerebrospinal Fluid; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Meningeal Neoplasms; Meninges; Middle Aged; Prostatic Neoplasms

We wished to assess the antigenic expression of primary lung tumors diagnosed as either carcinosarcoma or sarcoma in order to determine whether this information would be useful in distinguishing the two. We therefore immunohistochemically analyzed six pulmonary carcinosarcomas and five primary lung sarcomas for the presence of carcinoembryonic antigen (CEA), S100 protein, cytokeratin and vimentin using commercially available monoclonal and polyclonal antibodies on formalin fixed tissues. Six of six carcinosarcomas stained positively for cytokeratin while none of the sarcomas stained. In three carcinosarcomas both the carcinomatous and sarcomatous areas were positive while in three only the carcinomatous areas were positive. CEA staining was present in five carcinosarcomas and absent in all the sarcomas. CEA positivity was strong and not confined to those tumors with obvious gland formation. Staining for S100 protein was positive in two carcinosarcomas but only in those areas showing chondroid differentiation. Immunohistochemical staining for vimentin using two different monoclonal antibodies gave inconsistent results. We conclude that in differentiating between a carcinosarcoma and a sarcoma of the lung, immunohistochemical staining for both cytokeratin and CEA are useful with cytokeratin marginally preferable. The data indicate that carcinosarcoma of the lung, like that of the upper aerodigestive tract, expresses antigens suggesting both epithelial and mesenchymal differentiation.

Topics: Antigens, Neoplasm; Carcinoembryonic Antigen; Carcinosarcoma; Cell Transformation, Neoplastic; Diagnosis, Differential; Epithelium; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mesoderm; S100 Proteins; Sarcoma; Vimentin

Filed formalin-fixed paraffin blocks of 128 cases of epithelial neoplasms were selected for immunohistochemical study of keratin and vimentin expression. The results showed that 35.1% (45/128) of different carcinomas expressed vimentin. The immuno-positivity of vimentin in thyroid carcinomas, ovarian carcinomas, prostatic adenocarcinomas, pulmonary carcinomas and malignant mesotheliomas were 81.8%, 42.8%, 66.7%, 30.5% and 53.4%, respectively. Carcinomas of breast, kidneys, salivary glands, adrenal glands and nasopharyngeal carcinomas also showed various degrees of positive reaction. The results suggest that an immunohistochemical positive vimentin reaction does not exclude histopathological diagnosis of carcinomas. The significance and noticeable aspects of immunohistochemical methods in histopathological diagnosis are also discussed.

Topics: Carcinoma, Squamous Cell; Female; Humans; Intermediate Filaments; Keratins; Lung Neoplasms; Male; Mesothelioma; Neoplasms, Glandular and Epithelial; Ovarian Neoplasms; Prostatic Neoplasms; Thyroid Neoplasms; Vimentin

We are reporting the clinical and pathologic features of a primary, pulmonary, malignant germ cell tumor associated with a marked elevation of serum alpha-fetoprotein (38,427 ng/mL) and lactate dehydrogenase activity (756 U/L), in a 26-year-old female. This controversial, rare neoplasm has not been extensively discussed in the pathology literature. We emphasize the clinical importance of establishing this diagnosis in view of the favorable response to chemotherapy shown by malignant germ cell tumors.

Topics: Adult; Alkaline Phosphatase; alpha-Fetoproteins; Carcinoembryonic Antigen; Chorionic Gonadotropin; Chromogranins; Female; Humans; Immunohistochemistry; Keratins; L-Lactate Dehydrogenase; Lung Neoplasms; Membrane Glycoproteins; Mesonephroma; Microscopy, Electron; Mucin-1; Neoplasms, Germ Cell and Embryonal; S100 Proteins

In 17 cases of resected small cell carcinoma of the lung, there were 4 cases of central type and 13 cases of peripheral type. Histologic subtypes were classified into oat cell carcinoma (OAT), intermediate cell type (INT), and small cell carcinoma with large cell component (SC/LC). SC/LC was divided according to the criteria of Radice et al. Immunohistochemically, gastrin-releasing peptide (GRP) and neuron specific enolase (NSE) were used as markers for neuroendocrine cells, and keratin and secretory component (SC) were used as markers for epithelial and gland epithelial cells, respectively. Histologically, 4 cases of the central type were divided into 3 cases of INT and one case of SC/LC. Thirteen cases of the peripheral type were divided into 3 cases of OAT, 6 cases of INT, and 4 cases of SC/LC. SC/LC was more frequently seen in the peripheral type than in the central type. Immunohistochemically, there was no difference in the frequency of positive staining for GRP and NSE between the central and peripheral types, but positive staining for keratin and SC were more frequent in the peripheral type than in the central type. Three cases who survived more than 3 years were histologically divided into two cases of INT and one case of SC/LC. Immunohistochemically, these 3 cases showed positive staining for GRP or NSE, but also showed positive staining for keratin or SC. Our results showed that some of the peripheral type small cell carcinoma of the lung had histologic and immunohistochemical features which were different from those of typical small cell carcinoma. Long survival time after resection in some of the peripheral cases might be due to these features.

Topics: Aged; Aged, 80 and over; Carcinoma, Small Cell; Gastrin-Releasing Peptide; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Peptides; Phosphopyruvate Hydratase; Prognosis; Secretory Component

Sixty-seven cases of small cell lung carcinoma (SCLA) in Tri-Service General Hospital (TSGH) during the past 16 years were studied. For patients with extensive stage of disease, the mean survival time and 2-year survival rate were 7.2 months and 3.1% versus 13.4 months and 16.7% for patients with limited stage. A better prognosis was obtained by treatment with a combination of intensive chemotherapy and radiotherapy. Immunohistochemical studies were performed by the peroxidase-antiperoxidase method. The positive rates in descending order were bombesin (80%), synaptophysin (74.3%), neurofilament (68.6%), neuron-specific enolase (60%), low molecular weight cytokeratin (54.3%), high molecular weight cytokeratin (25.7%), chromogranin-A (22.9%), adrenocorticotrophic hormone (0). Seven cases were examined and found to be ultrastructure; only 3 cases were found to contain neurosecretory granules. We emphasize that electron microscopy is not necessary as a routine diagnostic procedure, while light microscopy should be employed whenever possible; the immunohistochemical study should be considered within this context.

Topics: Adrenocorticotropic Hormone; Adult; Aged; Bombesin; Carcinoma, Small Cell; Chromogranins; Female; Humans; Immunohistochemistry; Intermediate Filaments; Keratins; Lung Neoplasms; Male; Membrane Proteins; Microscopy, Electron; Middle Aged; Phosphopyruvate Hydratase; Survival Rate; Synaptophysin

A case of neuroendocrine lung tumor located beneath the pleura in a 71-year-old woman is reported. At autopsy, the tumor was found to have metastasized to the bones and liver without involving the hilar lymph nodes. Histologically, the tumor cells at the primary site and in the liver metastasis exhibited a carcinoid-like organoid structure, whereas pleomorphic giant cells were noted in the bone metastasis. The argyrophilic tumor cells were immunoreactive for neuron-specific enolase, chromogranin A, serotonin, calcitonin, calcitonin gene-related peptide, gastrin-releasing peptide, neuropeptide Y, gastrin, pancreatic polypeptide, glicentin, the alpha-subunit of human chorionic gonadotropin, keratin, epithelial membrane antigen, Leu M1 and carcinoembryonic antigen. Electron microscopy revealed abundant neurosecretory granules in the cytoplasm. This was considered to be a rare case of neuroendocrine lung tumor showing carcinoid-like histology at the primary site and large-cell transformation in bone metastasis.

Topics: Aged; Autopsy; Bone Neoplasms; Calcitonin; Calcitonin Gene-Related Peptide; Carcinoembryonic Antigen; Carcinoid Tumor; Cell Transformation, Neoplastic; Chorionic Gonadotropin; Chromogranin A; Chromogranins; Female; Giant Cell Tumors; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Membrane Glycoproteins; Microscopy, Electron; Mucin-1; Phosphopyruvate Hydratase; S100 Proteins; Serotonin

We report a 58-year-old woman with proliferating trichilemmal cyst (PTC), from which a spindle cell carcinoma arose. The tumor on her scalp had been removed at another hospital. Histological examination had revealed almost typical features of PTC. However, the case showed a partial transformation to spindle cell carcinoma, and transition zones between squamous epithelium and spindle cells were present. Three months after histologic examination, the patient came to us for the treatment of recurrent tumor. Despite surgical resection, the patient died as a result of distant metastases. Histologically, the recurrent tumor was composed of only spindle-shaped tumor cells. We describe the first example of this uncommon condition.

Topics: Carcinoma; Diagnosis, Differential; Epidermal Cyst; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Middle Aged; Skin Diseases; Skin Neoplasms

Mammary adenocarcinoma was diagnosed in a 3-year-old Landrace sow with prolonged infertility, anorexia and progressive emaciation after parturition. Gross examination confirmed a large tumour in the left anterior mammary gland with metastatic nodules on the pleura and in the parenchyma of lung. Microscopically, the tumour consisted mainly of solid adenomatous proliferations with numerous mitotic figures. Irregular glandular structures, solid nests of polygonal tumour cells without polarity and nests consisting of glandular, cribriform and solid portions were evident in dense fibrous stroma. Immunostaining revealed keratin in the tumour cells.

Topics: Animals; Carcinoma; Female; Immunohistochemistry; Keratins; Lung Neoplasms; Mammary Neoplasms, Animal; Swine; Swine Diseases

We examined pulmonary carcinomas with prominent sarcoma-like lesions both clinicopathologically and immunohistochemically. Grossly, two tumors had predominantly endobronchial growths, four bulky parenchymal growths, and two endobronchial, parenchymally mixed growths. In these eight patients, six tumors were completely resected, one patient was given irradiation only, and one patient died in the early postoperative period. On the basis of specific differentiation of the sarcoma-like lesions, the tumors were separated into three groups: two with "true" sarcoma differentiated into soft tissues such as striated muscle or osteoid tissue; three with a fibromatous sarcoma resembling atypical pseudosarcomatous stroma; and three with spindle cell carcinoma with evidence of epithelial differentiation. The prognosis was poor, and tumors with specific differentiation into rhabdomyosarcoma, chondrosarcoma, or spindle cell carcinoma progressed more rapidly than did those with a fibromatous sarcoma. Because the fibromatous sarcoma-like lesions were found to relate to a longer survival time for the patients, we wish to emphasize that a distinction of sarcomatous components should be made with regard to assessing the prognosis of pulmonary carcinoma with sarcoma-like lesions.

Topics: Aged; Carcinoembryonic Antigen; Carcinoma; Carcinosarcoma; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; Secretory Component; Vimentin

Small-cell lung cancer (SCLC) lines are distinguished from non-small-cell lung cancer (NSCLC) lines by their growth in floating aggregates, in contrast to the adherent monolayers formed by NSCLC cells in culture. Of 50 well-characterized SCLC lines recently described by the National Cancer Institute (NCI)-Navy Medical Oncology Branch, only four variant cell lines (SCLC-v) grew as adherent monolayers. One line, NCI-H446, was unique in growing long-term with coexisting floating and surface adherent subpopulations. We have physically segregated these two populations over many passages in vitro to enrich for relatively pure cultures of floating and adherent cells. No differences in c-myc expression, keratin pattern, or cytogenetic appearance were found between the adherent and floating sublines. However, expression of the neuroendocrine marker neuron-specific enolase in the floating cells was three times that found in the adherent cells. The floating subline also had much greater surface expression of neuroendocrine tumor antigens detected by monoclonal antibodies UJ13A and HNK-1, which have been recently shown to detect the neural cell adhesion molecule (NCAM) on SCLC cells. Two other adherent SCLC-v lines were also found to be unreactive with UJ13A and HNK-1, generalizing the association between NCAM expression and the growth of most SCLC cultures as floating aggregates. In conclusion, we have an interesting model to study expression of NCAM as related to the adhesive properties of SCLC cells.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Blotting, Northern; Blotting, Western; Carcinoma, Small Cell; Cell Adhesion; Cell Adhesion Molecules, Neuronal; Cell Division; Cytogenetics; Down-Regulation; Gene Expression; Humans; In Vitro Techniques; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase; Proto-Oncogene Proteins c-myc; Tumor Cells, Cultured

In a preliminary study for the development of an automated lung cancer cytology screening system utilizing both flow and image processing techniques, potential markers for the flow cytometric screening for carcinoma cells in sputum were analyzed. Immunostains were applied by the avidin-biotin peroxidase complex method, using antibodies to keratin (55-57 KD), TA-4 and SCC (antigens of squamous cell carcinoma of the human uterine cervix), neuron-specific enolase (NSE), gastrin-releasing peptide (GRP) and carcinoembryonic antigen (CEA), to carcinoma cells from 123 cases with lung cancer (35 with squamous cell carcinomas, 64 adenocarcinomas, 13 large cell carcinomas, 5 small cell carcinomas and 6 other histologic types) and to sputum cells from 113 cytologically negative cases (as controls). The positive rates were 60.1% for keratin, 34.8% for TA-4, 28.2% for SCC, 1.4% for NSE, 0.1% for GRP and 7.9% for CEA for carcinoma cells (P less than .05 for all) and 7.4% for keratin and 1.0% for SCC for sputum cells (P less than .05 for both). It was concluded that keratin is the most effective marker, not only for squamous cell carcinoma, but also for adenocarcinoma and large cell carcinoma. Since most small cell carcinoma cells in sputum have little or no cytoplasm, it is necessary to use an intranuclear marker to detect this histologic type.

Topics: Antibodies, Monoclonal; Automation; Biomarkers, Tumor; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mass Screening; Sputum

Routinely-fixed and Papanicolaou stained smears with the cytologic diagnosis of undifferentiated malignant neoplasm that had been prepared with cells obtained by fine needle aspiration biopsy (n = 7), pulmonary lavage (n = 5), or thoracentesis (n = 3) from 15 unselected patients were stained by an immunocytochemical technique to evaluate the presence of keratin proteins and the leukocyte common antigen (LCA). Commercially available, well-characterized monoclonal antibodies with specificities for keratin proteins and the leukocyte common antigen, and a streptavidin-biotin-horseradish peroxidase labelling method were used. Evaluation of the stained smears revealed the presence of one of the two antigens in material obtained from each patient, thus indicating the probable cell-lineage of the neoplastic cells. The specificity of the monoclonal antibody reagents used was further evaluated in routinely-fixed and stained cytologic material from 24 histologically confirmed carcinomas and 12 lymphomas. In conclusion, immunocytochemical techniques may be successfully applied to routinely processed archival cytologic smears to determine the antigenic profile of morphologically undifferentiated cells and therefore aid in the differential diagnosis of undifferentiated malignant neoplasms.

Topics: Adult; Aged; Antigens, CD; Antigens, Neoplasm; Carcinoma, Small Cell; Cell Membrane; Cytoplasm; Diagnosis, Differential; Female; Histological Techniques; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphoma; Male; Middle Aged; Staining and Labeling

Keratin and carcinoembryonic antigen immunoperoxidase staining were applied to lung cancer cells in paraffin sections of cell blocks from sputa and bronchial washings. The results correlate well with cytologic diagnosis. The large cell carcinomas are nonreactive with both antibodies.

Topics: Adenocarcinoma; Bronchi; Bronchoalveolar Lavage Fluid; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Histological Techniques; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Paraffin; Sputum

Thirty-three patients with small cell carcinoma of the lung were treated surgically, and immunohistochemistry of the cell differentiations was examined in detail. The overall 5-year survival rate was 38% and the rates in patients with stage I or stage III were 57% and 11%, respectively (P less than 0.05). Survival rates in patients with the oat cell type and intermediate type were 24% and 44%, respectively, but with no significant difference. This carcinoma seemed to originate from primitive multipotential stem cells, i.e., those of a neuroendocrine or epithelial nature. Histochemically and immunohistochemically, argyrophilic granules and neuron-specific enolase, neuroendocrine markers, were detected more frequently in the oat cell type rather than in the intermediate type. In contrast, keratin, epithelial membrane antigen, and carcinoembryonic antigen, epithelial origin markers, were present more frequently in the intermediate type than in oat cell type. However, the difference was significant only in case of detection of argyrophilic granules and the carcinoembryonic antigen (P less than 0.05). Our current recommendation is that surgical resection should be done in the earlier stage in both subtypes. A more favorable prognosis can be expected when adjuvant chemotherapy is prescribed.

Topics: Adult; Aged; Carcinoembryonic Antigen; Carcinoma, Small Cell; Cytoplasmic Granules; Female; Histocytochemistry; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; Phosphopyruvate Hydratase

Three cases of pulmonary blastoma (PB) were investigated microscopically with conventional stainings and immunohistochemically with monoclonal antibodies to cytokeratin, vimentin, desmin and neurofilament protein. The tumors differed in terms of morphology as well as of immunohistochemistry. Two were epithelial and mesenchymal mixed tumors, and the remaining one was a monophasic tumor of a typical blastemic character. The two mixed tumors also differed from each other. In one of them, the epithelial and mesenchymal component expressed cytokeratin and vimentin in a clear-cut manner without any transition. The other mixed tumor displayed a gradual epithelial-to-mesenchymal transition accompanied by a switch in the expression of cytokeratin and vimentin. The third tumor was of pure mesenchymal origin, expressing vimentin in the majority of cells and desmin in few cells. It is concluded that the PB is a morphologically and histogenetically heterogeneous tumor. Metaplastic changes may take place within a PB and make the recognition of embryogenesis more difficult.

Topics: Aged; Carcinoma, Squamous Cell; Desmin; Epithelium; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasms, Germ Cell and Embryonal; Vimentin

A series of 12 pulmonary lesions initially diagnosed as leiomyomatous in origin were reviewed. The original slides were analyzed for a series of histological features, the patients' charts were reviewed for clinical presentation and follow-up, and the usefulness of immunohistochemistry was assessed. Three groups were noted after analysis. Group 1, or benign lesions (three cases), all occurred in premenopausal female patients who remain alive and well. Group 2, or leiomyosarcomas (five cases), mostly occurred in older females, were thought to originate in the uterus, and resulted in death in four patients. In one of three cases of metastatic leiomyosarcoma, estrogen receptor was identified. Group 3 (4 cases) consisted of other diagnoses, including leiomyomatosis, fibrous histiocytoma, metastatic malignant melanoma, and metastatic synovial sarcoma. Immunohistochemistry was useful in separating nonsmooth muscle tumors and demonstrated muscle marker expression in all benign and most malignant smooth muscle lesions. Criteria are proposed for the diagnosis of pulmonary leiomyosarcoma, whether primary or secondary. In addition, if strict criteria are applied, the term "benign metastasizing leiomyoma" should be abandoned.

Topics: Actins; Adult; Aged; Desmin; Female; Humans; Immunohistochemistry; Keratins; Leiomyoma; Lung Neoplasms; Male; Middle Aged; S100 Proteins; Vimentin

In order to validate xenografted small cell lung carcinomas (SCLC) for biological studies, the authors established 12 lung neuroendocrine (NE) tumors (eight typical SCLC and four atypical NE tumors [ANE]) by heterotransplantation onto nude mice. Their characterization was performed using serial ultrastructural, enzymatic, and immunohistochemical methods on primary tumors and after xenografts. These were subclassified into epithelial (one), neuroendocrine (three), and multidifferenciated (eight) types. The phenotypic characters (cytokeratins, neurofilaments, neurone-specific enolase) and the proliferative rate (Ki 67 labelling) of original tumor were maintained until the last passage studied. Although further acquisition of subsets of cytokeratin or neurofilaments was observed in some cases, the authors could not detect any morphologic and/or biological spontaneous change comparable to those described in in vitro cell lines. In addition, ANE are not quite identical to variant subclasses described in vitro. The authors conclude that the stability of heterotransplanted SCLC is an advantage in further biological studies.

Topics: Animals; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Small Cell; Cytoplasm; Female; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Mice; Mice, Nude; Molecular Weight; Neoplasm Transplantation; Neurofilament Proteins; Phenotype; Phosphopyruvate Hydratase

Two unique cases of pulmonary carcinomas showing squamous and glandular differentiation and the production of excessive amounts of extracellular waxy eosinophilic material resembling amyloid are reported. Immunohistochemical studies showed strong staining of the neoplastic cells with antibodies directed at cytokeratin, epithelial membrane antigen, and carcinoembryonic antigen. Of particular interest was the simultaneous strong staining for S-100 protein and vimentin. The histology, ultrastructure, and immunohistochemical findings suggest focal myoepithelial differentiation in these mixed carcinomas and indicate an analogy to salivary gland neoplasms, particularly adenoid cystic carcinoma and dermal analog tumors.

Topics: Adenocarcinoma; Aged; Antigens, Neoplasm; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Extracellular Matrix; Female; Humans; Hyalin; Keratins; Lung Neoplasms; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; S100 Proteins; Vimentin

In this study antibodies specific for different intermediate-sized proteins (cytokeratins and neurofilaments) have been tested on a series of neuroendocrine (NE) lung tumors in order to evaluate their diagnostic validity. In particular we used a panel of polyclonal anti-neurofilament 200-kilodalton subunits whose reactivity against phospho-dependent epitopes was known. At least one NF subunit was constantly present and in all cases coexpression of cytokeratins and neurofilaments was confirmed. However, in cases of carcinoid tumor (CT) the results were homogeneous, while the cases of small cell lung carcinoma (SCLC) showed a much wider range of immunostaining. Our investigation confirms the hypothesis that the phosphorylation state is a significant determinant of immunohistochemical properties of neurofilaments. This might explain the large number of negative results obtained in previous investigations on NE tumors. The phosphorylation of neurofilaments may also be considered an indication of the degree of differentiation of the tumor.

Topics: Biomarkers, Tumor; Carcinoma; Carcinoma, Small Cell; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Phosphorylation

According to the World Health Organization histological classification of bronchial tumors, clear and giant cell carcinomas are two subtypes of large cell carcinoma. As clear and giant cells can also be observed in other types of bronchial carcinoma, we investigated the frequency of the finding of these cells in different histological types. The tumor size and degree of differentiation, the amount of necrosis and keratinization, and the presence of giant and clear cells were analyzed. Statistical analysis by X2 test showed (for all classified histological types of bronchial carcinomas, except small cell carcinoma) that: 1) larger tumors had a great quantity of giant cells (P less than 0.05; P less than 0.01), 2) large tumors had more clear cells (P less than 0.05; P less than 0.01) and 3) tumors with a greater amount of necrosis had a larger number of giant and clear cells (P less than 0.05; P less than 0.01). Findings of an identical cytological characteristic can cause some difficulty in determination of bronchial cancer.

Topics: Adenocarcinoma; Carcinoma, Bronchogenic; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Necrosis; World Health Organization

Flow cytometric DNA analysis using the anti-cytokeratin antibody was carried out in order to estimate more reliable measurement in single cell suspension obtained from solid tumors. It was difficult to detect a DNA aneuploidy with DI of 2.0 by one parameter analysis of DNA. Whereas it could be detected easily by using dual parameter analysis of cytokeratin and DNA. And also, the pattern of DNA multiploidy could be selected for cytokeratin positive cell population by gate analysis.

Topics: Antibodies; DNA, Neoplasm; Flow Cytometry; Humans; Keratins; Lung Neoplasms; Ploidies; Rectal Neoplasms; Tumor Cells, Cultured

In order to differentiate between malignant pleural mesothelioma and adenocarcinoma of the lung, the glycoconjugate profiles of 6 reactive mesothelial lesions, 23 mesotheliomas (17 epithelial, 1 desmoplastic, 2 biphasic, and 3 fibrous types), and 28 well-differentiated pulmonary adenocarcinomas were evaluated with the use of 8 lectins in addition to anti-carcinoembryonic, anti-keratin and anti-epithelial membrane antigen. Formalin-fixed, paraffin-embedded tissues were stained with the avidin-biotin peroxidase complex method. Reactions of wheat germ (WGA) and peanut (PNA) agglutinin with neuraminidase treatment lectins were positive in 5 of 6 (83%) and 3 of 6 (50%) cases, respectively, in reactive mesothelial lesions. Thirteen of 23 (57%) malignant mesotheliomas of the pleura showed a positive reaction for WGA and PNA with neuraminidase treatment; other lectins were low-positive, below 9%. In contrast, pulmonary adenocarcinomas showed positive reactions in 27 of 28 cases (96%) for PNA, 26 of 28 (93%) for Ricinus communis (RCA-I), 25 of 28 (89%) for WGA, and 22 of 28 (79%) for succinylated WGA (SucWGA). The findings suggest that malignant pleural mesothelioma and pulmonary adenocarcinoma have consistent and distinct glycoconjugate profiles, and that stains for RCA-I and SucWGA may be useful for differential diagnosis.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Diagnosis, Differential; Histocytochemistry; Humans; Keratins; Lectins; Lung Neoplasms; Membrane Glycoproteins; Mesothelioma; Mucin-1; Pleural Neoplasms

Eight primary carcinomas of the lung with a prominent spindle-cell sarcomatoid component were studied by immunocytochemical staining and electron microscopy. The eight tumors were indistinguishable by conventional light microscopy, with the exception of one unusual neoplasm that followed multiple pathways of differentiation with elements of squamous cell carcinoma, rhabdomyosarcoma, chondrosarcoma, and an undifferentiated spindle-cell population. Reticulin fiber production by individual spindle cells and a sharp demarcation of the carcinomatous and sarcomatoid domains by light microscopy were not useful differentiating features. Three of the eight tumors exhibited keratin expression in both the carcinomatous and spindle-cell components. Both immunocytochemical and electron microscopic analyses were required to detect epithelial differentiation, as in one case keratin was identified only by immunocytochemical staining and in another only by ultrastructural examination. Epithelial differentiation was undetectable in the sarcomatoid component of five tumors, and in one case immunoreactive myoglobin was identified in spindle cells; skeletal muscle differentiation was confirmed ultrastructurally. We propose that pulmonary carcinomas exhibiting evidence of epithelial differentiation in a sarcomatoid component be termed spindle-cell carcinomas and that those biphasic tumors exhibiting mesenchymal differentiation into specific tissues, such as neoplastic bone, cartilage, or striated muscle, or lacking epithelial differentiation by light microscopy, immunocytochemistry, and electron microscopy be classified as carcinosarcomas. This distinction may ultimately be unnecessary, because these two tumors may represent different points along a morphologic and biologic continuum.

Topics: Carcinoembryonic Antigen; Carcinoma; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Microscopy, Electron; Sarcoma

This report describes three cases of intrapulmonary fibromas which are histologically identical to localized fibrous tumors of pleura (localized fibrous mesothelioma). Morphologically these tumors are characterized by a haphazard proliferation of cytologically bland spindle cells separated by variable amounts of wavy hyalinized collagen. Entrapped bronchiolar and alveolar epithelium is common. These spindle cells lack expression of cytoplasmic keratin, S-100 protein, desmin, and epithelial membrane antigen, but are strongly decorated for intracellular vimentin. The clinical behavior, differential diagnosis, and histogenesis of these lesions are discussed.

Topics: Aged; Aged, 80 and over; Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Mesothelioma; Middle Aged; Necrosis; Periodic Acid-Schiff Reaction; Vimentin

In order to improve the accuracy of diagnosis and subtyping of pulmonary adenocarcinomas, immunohistochemical studies were carried out on 105 adenocarcinomas of the lung procured from both surgery and autopsy. Avidin-biotin-peroxidase complex methods were used for identifying keratin, vimentin, carcinoembryonic antigen (CEA), and secretory component (SC) on deparaffinized tissue sections. Keratin was positive in 29% of well differentiated adenocarcinoma, significantly lower than in moderately or poorly differentiated adenocarcinoma. Likely, vimentin was positive in 27% of well differentiated adenocarcinoma, significantly lower than in moderately or poorly differentiated adenocarcinoma. SC was positive in 66% of well differentiated adenocarcinoma, significantly higher than in moderately or poorly differentiated adenocarcinoma. In the subtyping of well differentiated adenocarcinomas, keratin showed higher positive results in the bronchial surface epithelial, goblet cell, and bronchial gland types than in the Clara cell or type II alveolar epithelial cell type. These findings suggest that immunoperoxidase stains for keratin, vimentin, and SC may be useful for determining the degree of differentiation of adenocarcinomas of the lung as well as for subtyping of well differentiated pulmonary adenocarcinomas.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoembryonic Antigen; Humans; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Lung Neoplasms; Secretory Component; Vimentin

An extremely rare solitary mast cell tumor of the lung was studied histologically, immunohistochemically, and ultrastructurally. The histologic features of the tumor included nodular growth of well-differentiated mast cells and clear cells with no granules. The current case is the third case of a solitary mast cell tumor (granuloma) of the lung in the literature. Clinicopathologic features of this tumor are compared with the other two cases reported previously in the international literature, and the nature of the clear cells is discussed.

Topics: alpha 1-Antitrypsin; Antigens, Neoplasm; Apoproteins; Diagnosis, Differential; Granuloma; Humans; Keratins; Lung Neoplasms; Male; Mast Cells; Mast-Cell Sarcoma; Membrane Glycoproteins; Middle Aged; Mucin-1; Muramidase; Neoplasm Proteins; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants

The molecular forms of keratin in small cell lung cancer (SCLC) cell lines and tumors were examined with antikeratin monoclonal antibodies. Immunostaining of SCLC by antikeratin antibody and examination by fluorescence microscopy indicates population heterogeneity in keratin content. Intensity of immunostaining is often weak. However, polyacrylamide gel electrophoresis and immunoblotting reproducibly demonstrate the presence of keratin and allow analysis of the keratin subtypes. The finding of keratin subtypes closely associated with the development of keratinizing epithelium (the 68 kDa basic keratin) in SCLC was unexpected in a tumor that is regarded as poorly differentiated. The cytoskeletal composition of SCLC suggests the presence of a heterogeneous population with a significant proportion of cells expressing highly differentiated epithelial properties.

Topics: Carcinoma, Small Cell; Electrophoresis; Fluorescent Antibody Technique; Humans; Keratins; Lung Neoplasms; Tumor Cells, Cultured; Vimentin

The expression of cytokeratins (CKs) in human lung cancer was studied using chain-specific monoclonal antibodies to CKs 4, 7, 8, 10, 13, 18, and 19. When applied to adenocarcinomas (ACs) of the lung, high levels of CKs 7, 8, 18, and 19 were detected in all tumors, while CK 4 was found in high concentrations in some ACs. CK 10 and 13 were completely absent, or only present in low numbers of cells. Small cell lung cancers (SCLCs) and lung carcinoids contained CK 18 and sometimes 8 and 19, but no CK 7 in most cases. Three out of four tumors, histologically classified as SCLC, and expressing CK 7 in a variable number of cells were found by electron microscopic studies to contain regions with AC and/or squamous cell carcinoma (SQC) differentiation. The monoclonal antibody specific for CK 7 can therefore possibly help to distinguish AC differentiation within SCLC. CKs 10 and 13 were completely absent in SCLCs and lung carcinoids, while few CK 4-positive cells were found in some SCLCs and in one lung carcinoid. Within SQCs the monoclonal antibodies revealed a pronounced heterogeneity in CK expression. CKs 4, 7, 8, 10, 13, 18, and 19 could be detected, although not evenly distributed among all tumor cells. Highly differentiated SQCs expressed high levels of the CKs specific for squamoid differentiation, i.e., CKs 4, 10, and 13 in variable numbers of cells. With decreasing histologically detectable SQC differentiation these markers were gradually lost, while the number of cells containing CKs 7, 8, 18, and 19 increased. Application of this panel of monoclonal antibodies can therefore distinguish not only the main subtypes of lung cancer, but can also indicate the degree of differentiation and the degree of heterogeneity. These findings can be used as a diagnostic aid in lung tumor pathology, which may have an impact on treatment and prognosis.

Topics: Antibodies, Monoclonal; Carcinoma; Cytoskeleton; Electrophoresis, Polyacrylamide Gel; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms

An in situ hybridization technique was applied to detect expression of keratin mRNAs in xenotransplanted human tracheobronchial epithelium and lung carcinomas. Tissues from eight tracheas repopulated with cells from five different noncancerous donors and 15 squamous cell carcinomas were used. Using a K6 (56 kd) human keratin cDNA (KA-1) and a K14 (50 kd) cDNA (KB-2) as probes, radiolabeled by nick-translation with 3H-dATP/TTP, the specificity and significant differences in the levels of silver grains on various epithelial lesions in formalin-fixed, paraffin-embedded tissue sections were demonstrated. In situ hybridization with either KA-1 or KB-2 probe showed similar localization of silver grains in all histologic types in consecutive tissue sections. In xenotransplanted tracheobronchial epithelia, very few grains were seen over cells of simple, pseudostratified, or stratified epithelia two to three cell layers thick. Nonkeratinizing stratified hyperplastic epithelia of more than three cell layers showed uniform localization of numerous grains throughout the lesions. In contrast, epidermoid metaplasias exhibited a dense and localized pattern of grains on the basal and parabasal cell layers with a decrease in grain density toward the surface layers. Carcinoma cells from bronchogenic squamous cell carcinomas showed a higher density and more uniform localization of grains. Well-differentiated carcinoma cells contained more keratin mRNAs than moderately to poorly differentiated carcinoma cells. This evidence obtained with the KA-1 and KB-2 probes demonstrates the different localization patterns of keratin mRNAs in different epithelial lesions. In addition, the levels of mRNA expressed show a positive correlation with the degree of squamous differentiation. It was of particular interest that an ordered program of keratin mRNA expression proportional to the level of cellular differentiation was observed in epidermoid metaplasias. Both of these probes serve as keratinization markers of human tracheobronchial epithelial lesions.

Topics: Animals; Bronchi; Carcinoma, Bronchogenic; Carcinoma, Squamous Cell; DNA; Epithelium; Humans; Keratins; Lung Neoplasms; Mice; Mice, Nude; Nucleic Acid Hybridization; RNA, Messenger; Trachea; Transplantation, Heterologous

N-Propyl-N-nitrosourea is a strong leukemogen that induces myelogenic leukemia in Donryu rats and thymic lymphoma in F344 rats when administered in drinking water. In the present study, a single or multiple doses of PNU (total 500 mg/kg body weight) was given to young male and female F344 rats via a stomach tube. The results demonstrated that the percentage of tumor-bearing rats was 100% in all PNU-treated male groups, while that of the control group was 46%. Predominant tumors induced by PNU in male rats were lung adenoma/adenocarcinoma followed by peritoneal mesothelioma, and forestomach papilloma. In females, the tumor incidence of PNU-treated groups varied between 58% and 92% while that of the control group was 42%. Although pituitary tumor was the most frequent tumor in PNU-treated female rats, it was thought to be spontaneous since its incidence in each experimental group was not statistically different from that of the control group. Lung tumors and forestomach papillomas were also induced by PNU in female rats. No thymic lymphoma, however, was found in any of the PNU-treated groups of either sex. Lung tumors developed in almost all PNU-treated male rats and in about one-third of PNU-treated female rats. Mesothelioma was induced only in male rats, and its incidence depended on the treatment schedule. Induced mesotheliomas were extensively examined histologically, histochemically, immunohistochemically, and electron microscopically.

Topics: Animals; Female; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Mesothelioma; Nitrosourea Compounds; Peritoneal Neoplasms; Rats; Rats, Inbred F344; Vimentin

Ten examples of giant cell carcinoma of the lung were examined by immunohistochemistry for expression of keratin and vimentin intermediate filaments and for epithelial membrane antigen (EMA). Six cases were also examined electron microscopically. Keratin expression and, to a lesser extent, EMA immunoreactivity were reduced in comparison with better differentiated forms of lung carcinoma. Vimentin expression was increased, often taking the form of strong paranuclear staining. This may correspond to dense paranuclear aggregates of intermediate filaments seen ultrastructurally. Desmosomes were absent or sparse in most tumours. We propose that giant cell carcinoma arises by a process of dedifferentiation. The resulting loss of epithelial features gives rise to neoplastic cells which have features in common with some forms of sarcoma.

Topics: Adenocarcinoma; Antigens; Carcinoma; Cell Differentiation; Cell Transformation, Neoplastic; Desmosomes; Humans; Keratins; Lung Neoplasms; Membrane Glycoproteins; Microscopy, Electron; Microvilli; Mucin-1; Vimentin

Topics: Adenocarcinoma; Humans; Keratins; Lung Neoplasms

A panel of human lung carcinoma lines was studied with respect to hormone production and intermediate filament expression to distinguish between endodermal and neuroendocrine differentiation. An index of the degree of neuroendocrine differentiation of each line was derived from the presence or absence of hormone production, cytokeratins, neurofilaments and an embryonic endodermal cell marker, which allowed identification of three groups showing high, intermediate or low neuroendocrine expression. This grouping correlated well with the in vitro radiosensitivity of the lines, those expressing pure neuroendocrine features being significantly more radiosensitive than those with an endodermal phenotype, with the intermediate group having intermediate sensitivity. Use of such an index might predict those patients likely to benefit from the use of radiotherapy in their management.

Topics: Adenocarcinoma; Adrenocorticotropic Hormone; Calcitonin; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Line; Humans; Intermediate Filaments; Keratins; Lung Neoplasms; Radiation Tolerance; Vasopressins

An immuno-histological study of metastatic adenocarcinoma has revealed the following results. Metastatic adenocarcinomas of the lymph-node of pulmonary and colonic origin were positive for CEA and negative for lysozymes, and those from gastric, pancreatic, and gallbladder tumors were positive CEA and lysozymes, and those from gastric and pancreatic tumors were positive for the secretory component. The prostate specific antigen was exclusively positive for metastatic prostatic adenocarcinoma with a low frequency and prostate acid phosphatase had many false positive results. Thyroglobulin was found to be positive only to colloid. Lactalbumin showed no specificity to metastatic breast adenocarcinoma. For achieving the final diagnosis of a primary tumor, its location in lymph nodes, the clinical history and the results of other examinations must also be taken into consideration.

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoembryonic Antigen; Colonic Neoplasms; Humans; Keratins; Lactalbumin; Lung Neoplasms; Lymphatic Metastasis; Muramidase; Neoplasms, Unknown Primary; Secretory Component

Sclerosing hemangioma of the lung (SHL) was investigated immunohistochemically, histochemically and ultrastructurally with reference to cellular components associated with the histologic pattern: cuboidal cells in the papillary type, round cells in the solid type, flat cells in the hemorrhagic type and stromal cells in the sclerotic type. Immunohistochemically, cuboidal cells were positive for CEA, cytokeratin and EMA, whereas other cells were positive for EMA and vimentin. Immunoreactive factor-VIII-related antigen was confined to endothelial cells. Histochemically, cuboidal cells displayed alkaline phosphatase activity, but round cells showed ATPase activity. However, in spite of these different histochemical and immunohistochemical properties, morphological continuity was clearly revealed in immunostained sections; direct connection of spaces lined by cuboidal and flat cells, direct contact between cuboidal and stromal cells, and EMA expression of round cells associated with luminal structures were evident. Ultrastructurally, cuboidal cells were like alveolar cells. Flat and stromal cells showed microvillous protrusions and a discontinuous basement membrane, but some cells contained lamellar bodies. Solid cellular sheets consisted of various cells intermediate between cuboidal and flat or stromal cells. Direct apposition among these cells was evident. This morphological continuum confirms that each of these cell types are components of SHL as a whole. SHL may thus be merely sclerotic hemorrhagic alveolar cell tumor.

Topics: Adenosine Triphosphatases; Aged; Alkaline Phosphatase; Carcinoembryonic Antigen; Factor VIII; Female; Histiocytoma, Benign Fibrous; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Membrane Glycoproteins; Microscopy, Electron; Middle Aged; Mucin-1; Vimentin

Malignant cell lines related to mouse lung alveologenic carcinoma have been established from urethane-induced tumors and after in vitro spontaneous transformation of preneoplastic cell lines. Both the chemically and spontaneously transformed cell lines formed invasive, poorly differentiated carcinomas with secondary lung deposits when implanted subcutaneously in immune-suppressed mice. They differed from the related preneoplastic cell line in coordinately exhibiting anchorage-independent growth, reduced epidermal growth factor receptor activity and absence of pericellular fibronectin. These data suggest that similar molecular events may occur in type 2 pneumocyte-related cells in order to generate mouse lung alveologenic adenomas and carcinomas by both spontaneous and chemical carcinogen induction mechanisms. A reduced level of pericellular fibronectin was also demonstrated in an in situ compressive urethane-induced mouse lung adenoma. Loss of pericellular fibronectin may therefore be an early and persistent phenotypic alteration during transformation to the alveologenic adenoma and carcinoma.

Topics: Adenoma; Animals; Cell Line, Transformed; Cell Transformation, Neoplastic; Female; Fibronectins; Keratins; Lung Neoplasms; Mice; Mice, Inbred BALB C; Pulmonary Alveoli; Urethane

Topics: Carcinoembryonic Antigen; Carcinoma, Small Cell; Humans; Keratins; Lung Neoplasms

The usefulness of cell lines in the study and prediction of the clinical behaviour of lung cancer is still a matter of debate. However, lung tumour cell cultures have been of value in investigations concerning molecular and cell biological aspects of these neoplasms. Especially in the examination of characteristics specific for the main types of differentiation (squamous cell carcinoma, adenocarcinoma, small cell carcinoma), in vitro studies have been most important. Twenty eight lung cancer cell lines were cultured for up to four years, and were examined at regular intervals for their intermediate filament protein (IFP) expression patterns using a panel of cytokeratin (CK) and neurofilament (NF) antibodies. These studies showed that the classic type of small cell lung cancer (SCLC) cell lines contain CKs 8, 18, and occasionally CK 19, while the variant-type SCLC cell lines generally express no CKs but can contain NFs. Non-SCLC cell lines, such as squamous cell carcinoma and adenocarcinoma cell lines, contain CKs 7 (in most cases), 8, 18 and 19. In one variant SCLC cell line and in one adenocarcinoma cell line CKs 4, 10 and 13, characteristic of squamous cell differentiation, were found. Although most cell lines have remained stable with respect to growth characteristics and IFP expression patterns, five lung cancer cultures exhibited a transition from one cell type to another, paralleled by changes in IFP expression. Progressions from classic to variant SCLC cell lines have been observed, next to conversions from variant SCLC to cell lines re-expressing cytokeratins. In some cases this resulted in a coexpression of CKs and NFs within a cell line and even within individual tumour cells. These results strongly support the earlier finding that CK expression in SCLC cell lines is a reliable marker for the classic type of differentiation, while the absence of CKs and the presence of NFs marks the variant type of differentiation. Our results are discussed in view of previous histological findings.

Topics: Carcinoma, Small Cell; Cell Line; Humans; Immunoblotting; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Microscopy, Fluorescence; Microscopy, Phase-Contrast; Tumor Cells, Cultured

A panel of seven monoclonal antiepithelial antibodies of different specificities, including anticytokeratin, human milk fat globule membrane, C, and carcinoembryonic antigen (CEA) were used with the alkaline phosphatase-antialkaline phosphatase (APAAP) immunostaining technique to determine their value in the differentiation between benign and malignant mesothelial cells and lung carcinoma in histological preparations. The anticytokeratin antibody reacted strongly with all cases of reactive mesothelium, mesothelioma, and lung carcinoma. Antibodies to human milk fat globule membrane and the Ca antigen stained mesothelioma and carcinoma and 43% of cases of reactive mesothelium. Staining for carcinoembryonic antigen was not detected in reactive mesothelium or mesothelioma, but was present in most of the lung carcinomas. CEA seemed to be the single most useful marker in distinguishing carcinoma from mesothelioma in that a positive reaction for CEA would indicate carcinoma rather than mesothelioma.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Carcinoembryonic Antigen; Carcinoma; Diagnosis, Differential; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Membrane Proteins; Mesothelioma; Mucin-1

Sections of primary lung carcinomas, lung metastases, mesotheliomas, and lung metastases of some rare mesenchymal tumors were incubated with different cytokeratin (CK), vimentin, desmin, and tissue polypeptide antigen (TPA) antibodies and with antibodies reactive with different hormones (ACTH, PTH, alpha-HCG, Calcitonin CT), CEA, carcinoma-associated antigen (CA1), secretory component (SC), neuron-specific enolase (NSE), alpha-1-antitrypsin (alpha-1-AT), lysozyme (lyso), and S-100 protein (S 100). CK antibodies derived from a 49 kD (reactive with simple epithelia [SE]) and a 67 kD CK polypeptide fraction (reaction with complex epithelia [CE] were useful differentiation markers for the four major groups of lung carcinomas. In one half of small cell carcinomas a positive reaction with NSE antibodies was found. S 100 and SC were good markers for papillary and bronchioloalveolar adenocarcinomas, whereas CEA was less important because of its reactivity with different types of lung carcinomas. To discern clear cell carcinomas of lung and renal origin a positive reaction with vimentin antibodies (some renal but not lung types) and with CA1 (no renal but all lung types) seemed to be useful. All hormone antibodies were of no importance as markers for difficult differential diagnosis, because positive reactivities were found in cases from every major carcinoma group. In addition, a Ca2+-activated adenosine triphosphatase (ATPase) was found in mesotheliomas but not in papillary adenocarcinomas.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Differentiation; Desmin; Diagnosis, Differential; Histocytochemistry; Hormones; Humans; Immunologic Techniques; Keratins; Kidney Neoplasms; Lung Neoplasms; Mesothelioma; Muramidase; Neoplasm Metastasis; Peptides; Phosphopyruvate Hydratase; Pleural Neoplasms; S100 Proteins; Secretory Component; Tissue Polypeptide Antigen; Vimentin

Topics: Adenocarcinoma; Breast Neoplasms; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cytoskeleton; Diagnosis, Differential; Epithelium; Female; Gastrointestinal Neoplasms; Humans; Intermediate Filament Proteins; Intermediate Filaments; Keratins; Lung Neoplasms; Neoplasms; Skin Neoplasms; Urinary Bladder Neoplasms; Uterine Cervical Neoplasms

In order to investigate the intermediate filament protein content of hormone producing lung tumor cell cultures a panel of 16 different cytokeratin antisera were tested using immunocytochemical and biochemical techniques on lung carcinoma cell cultures from different origin. These included three cell cultures derived from small cell lung carcinoma, two large cell carcinoma cell cultures, and two cell cultures derived from squamous cell carcinomas. Flow cytometric analysis of the cell cultures demonstrated that all cell lines examined were aneuploid with DNA-indices ranging from 1.7 to 3.1 rimes the DNA-content of normal human lymphocytes. In both immunofluorescence and immunoperoxidase techniques six out of seven cell cultures reacted with most of the cytokeratin antisera used in a filamentous manner, while a large cell carcinoma cell culture did not react with any of the cytokeratin antisera used. None of the cell cultures examined reacted with the antibodies to neurofilament proteins, suggesting that none of these (neuro)hormone producing cell cultures were of neural origin. All cell cultures which were growing as adherent cell cultures did express vimentin. The cell culture that grew with cells floating in aggregates did not express this intermediate filament protein while a subline which did attach, expressed vimentin. This findings strongly indicates the relation between growth pattern in vitro (floating vs. adherent) and the expression of vimentin. No reaction was found with antisera to desmin and GFAP. The presence of cytokeratins and vimentin in most cell cultures could be confirmed using one- and two-dimensional gel electrophoresis. Cytokeratins 7, 8, 18 and 19 were most commonly present.

Topics: Carcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Line; Cytoskeleton; Electrophoresis, Polyacrylamide Gel; Flow Cytometry; Fluorescent Antibody Technique; Histocytochemistry; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Intermediate Filaments; Keratins; Lung Neoplasms; Neurofilament Proteins

Lung cancers were investigated for their heterogeneity as expressed by their immunoreactivity for cytokeratins and neurofilament proteins, as well as for the neuroendocrine differentiation antigen MOC-1. Using broadly cross-reacting antibodies, cytokeratins were detected in nearly all cases of lung carcinomas. Keratinization could be detected only in cases of moderately to well-differentiated squamous cell carcinoma (SQC) using a monoclonal antibody to cytokeratin 10, while a monoclonal antibody reactive with cytokeratin 18, and specific for glandular epithelia, reacted with adenocarcinomas, small cell lung carcinomas (SCLC), and lung carcinoids. In SQC this antibody could detect non-squamous cell differentiation, showing increasing numbers of positive cells with decrease of histologically detectable SQC differentiation. Cells positive for neurofilaments were demonstrated in some of the poorly differentiated SQCs and in some of the cases of SCLC, possibly representing the variant type of SCLC. Also in some of the lung carcinoids neurofilament proteins were present, colocalizing with cytokeratins. MOC-1 was present in all SCLC and lung carcinoids. This antibody could also detect neuroendocrine differentiation in all combined small cell carcinomas, in one poorly differentiated adenocarcinoma, and in about 30% of the poorly differentiated SQCs. Therefore, lung cancer heterogeneity can be detected using a panel of well-defined antibodies to intermediate filaments in combination with the MOC-1 antibody. The use of these antibodies in diagnosis can have prognostic significance and can lead to a more selective therapeutic approach.

Topics: Adenocarcinoma; Antibodies; Antigens, Neoplasm; Carcinoid Tumor; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cross Reactions; Cytoskeleton; Histocytochemistry; Humans; Immunoenzyme Techniques; Intermediate Filaments; Keratins; Lung Neoplasms

A cultured small cell lung cancer cell line (Lu-134-B-S) established from a xenotransplanted tumor in a nude mouse, which had originated from a primary focus of small cell lung cancer, showed morphological changes when the medium was changed from RPMI 1640 supplemented with 10% fetal calf serum to RPMI 1640 supplemented with 10% delipidized fetal calf serum. That is, it consisted of "classic" small cells in the former medium, but after eight passages in the latter medium many cells became squamous cells, possessing abundant eosinophilic cytoplasm and intercellular bridges. Immunohistochemically, they reacted to antikeratin and antiinvolucrin antibodies. Electron microscopically, well developed desmosomes and associated tonofibrils were noted, and electrophoretically, the amount of medium (Mr 57,000 and 59,000) and large-sized (Mr 67,000) keratins were found to increase with the change of the medium. These changes reversed to the original small cell morphology within 4 weeks after addition of vitamin A (retinoic acid) to the medium. These findings suggested that deficiency of vitamin A caused the change of the cell from small to squamous cell and vice versa.

Topics: Aromatic-L-Amino-Acid Decarboxylases; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Line; Creatine Kinase; Culture Media; Humans; Isoenzymes; Keratins; Lung Neoplasms; Microscopy, Electron; Phosphopyruvate Hydratase; Vitamin A

Monoclonal antibody based immunohistochemistry is a very powerful tool for the establishment of a pathological diagnosis of lung cancer. Applying a panel of intermediate filament antisera and an antibody recognizing neuroendocrine differentiation we have tested about 240 human lung tumors and 15 human lung tumor cell lines. Our results can be summarized as follows: a differential diagnosis between neuroendocrine and non-neuroendocrine lung tumors can be obtained by the application of the monoclonal antibody MOC-1 directed against neuroendocrine antigens. Immunohistochemistry can lead to a better recognition of lung tumor heterogeneity within the established histologies. Examples of this phenomenon are: the presence of neuroendocrine and/or neural components within non-neuroendocrine tumors. The presence of squamous cell or adenocarcinomatous differentiation in non-SCLC can be detected by chain specific anti-cytokeratin antibodies. The degree of differentiation towards the variant type within SCLC can be detected by the monoclonal antibody directed against neurofilaments. lung cancer cell lines can serve as an in vitro model for immunohistochemical studies on different lung cancer subtypes.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Differentiation; Cells, Cultured; Diagnosis, Differential; Humans; Intermediate Filaments; Keratins; Lung Neoplasms

Pleural mesotheliomas and peripheral pulmonary adenocarcinomas were evaluated by immunohistochemistry at two institutions (Yale University School of Medicine and the City of Hope National Medical Center). Twenty-three mesotheliomas, 11 with ultrastructural verification, and 24 pulmonary adenocarcinomas were studied. Antikeratin, anticarcinoembryonic antigen, anti-human milk fat globule related antigen, MC-1, B72.3, and Leu M-1 antibodies were employed. The immunohistochemistry of each case prepared at one institution was reviewed at the other. As groups, the two tumor types were distinguishable using this antibody panel. Essentially, mesotheliomas were keratin positive only. The adenocarcinomas were positive for CEA, MFG, B72.3, and Leu M-1. However, there were some ambiguities presented by the immunohistochemistry. The higher molecular weight keratins were found in most but not all mesotheliomas and in only a few adenocarcinomas. Lower molecular weight keratins were positive not only in all the mesotheliomas, but also in nearly all the adenocarcinomas, but also in a tumor determined by electron microscopy to be mesothelioma. Preabsorption of CEA decreased the sensitivity for adenocarcinoma, but did not change the positivity of the putative mesothelioma. B72.3 and Leu M-1 were specific for adenocarcinoma, but were found in only half of them. The MFG was apparently specific for adenocarcinoma, but the findings were difficult to interpret. At the level of individual cases, greater sensitivity of separation can be achieved by combining the results of two or more antibodies, but the lack of a detectable specific antigen in mesothelioma continues to make some cases difficult to evaluate by immunohistochemistry alone.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoembryonic Antigen; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma; Microscopy, Electron; Pleural Neoplasms

The reaction patterns in 80 adenocarcinomas of the lung were examined with PAP-method using a monoclonal antibody against keratin and one against carcinoembryonic antigen (CEA) and a polyclonal antiserum against CEA. Almost all tumors showed a positive reaction to the antibodies which, however, varied quantitatively. Even though a reliable correlation of positive immunohistochemical reaction with the different light microscopical types was not possible according to WHO subtypes and degrees of differentiation, specific localization of the reaction within the tumor cells was seen with increasing differentiation. There was no correlation between the immunohistochemical reactions and 14 clinically measured plasma CEA levels. The plasma CEA level not only depends on CEA production by the tumor but also on other factors.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms

Three cases of "dedifferentiated" chordoma arising in the sacrococcygeal region are presented. In all three cases, the "dedifferentiated" component arose de novo in conjunction with conventional chordoma. Two of these patients, whose tumors had a prominent malignant fibrous histiocytoma (MFH) component, died within 6 months of diagnosis. Both patients had lung metastases, one of which was histologically documented to be MFH. The third patient, whose initial tumor contained osteosarcoma, died 76 months after diagnosis and multiple recurrences. Most notable in this case was the absence of the "dedifferentiated" component (in this instance, osteosarcoma) in all of the local recurrences as well as the lung metastases. These were composed exclusively of conventional chordoma. None of the patients had a previous history of radiation therapy. The immunohistochemical staining pattern of conventional chordoma was similar to that of previous reports, where the epithelial-like cells stained for cytokeratin and epithelial membrane antigen. In addition, they stained for alpha-1-anti-chymotrypsin and vimentin. These latter two markers were also identified in the "dedifferentiated" component. As with "dedifferentiated" chondrosarcomas and liposarcomas, "dedifferentiation" in a chordoma usually portends an accelerated clinical course.

Topics: Adult; Aged; Chordoma; Histiocytic Sarcoma; Histocytochemistry; Humans; Keratins; Lung Neoplasms; Male; Membrane Proteins; Mucin-1; Neoplasm Recurrence, Local; Osteosarcoma; Sacrococcygeal Region

A 26-year-old woman was operated on for a bulky tumor in the sacral region; she died of massive local tumor recurrence and pulmonary metastases 3 months later. Most of the original tumor showed a highly cellular spindle-cell sarcoma compatible with a fibrosarcoma of a high grade of malignancy. In a few small areas of the tumor, a chordoma-like pattern surrounded by growth of spindle-cell sarcoma was found. The spindle-cell component exhibited vimentin positivity in all tumor cells, but many cells were also cytokeratin-positive. The chordoma-like areas showed cytokeratin in all tumor cells. The chordoma-like areas, but not the spindle-cell areas also were positive for epithelial membrane antigen and S-100 protein. This case indicates that the sarcomatous change associated with chordoma may contain keratins as a sign of epithelial differentiation, and may thus represent sarcomatous transformation of chordoma cells, rather than a coincidental soft-tissue sarcoma or collision tumor.

Topics: Adult; Chordoma; Female; Fibrosarcoma; Humans; Keratins; Lung Neoplasms; Membrane Proteins; Mucin-1; Pelvic Neoplasms; S100 Proteins; Sacrococcygeal Region; Sarcoma; Vimentin

Synaptophysin is an integral membrane glycoprotein with an Mr of 38,000 that occurs in the small, clear vesicles present in neuronal cells and tumors as well as in pancreatic islet cells and various neuroendocrine (NE) carcinomas. We found that synaptophysin is also expressed in normal NE cells of the lungs of newborn rabbits and mice as well as of human fetuses. In bronchial ganglion cells and in nerves, synaptophysin is coexpressed with neurofilament proteins (NFPs), whereas in solitary NE cells and in at least some of the neuroepithelial bodies (NEBs) of the bronchial mucosal lining, synaptophysin coexists with cytokeratins. We also studied a series of NE neoplasms of the lung covering the entire spectrum of differentiation (i.e., from carcinoids to small-cell NE carcinomas), and found that synpatophysin was present in the majority of them. In these tumors, synaptophysin was invariably coexpressed with cytokeratin filaments and desmoplakin, as well as, occasionally, with NFP. Synaptophysin was identified throughout, the whole range of these NE neoplasms, i.e., from benign to low-grade to aggressive and rapidly metastasizing carcinomas; its presence was unaffected by the highly variable expression of serotonin and/or neuropeptides in these neoplasms, and was unrelated to the presence or absence of associated endocrine syndromes. Our findings indicate that synaptophysin occurs in the neural as well as in the epithelial components of the dispered NE system of the lung as well as in the majority of NE neoplasms of this organ, and that the expression of this protein is therefore independent of the cytoskeletal characteristics and other differentiation features of both normal and transformed NE cells of the lung. We emphasize the value of synaptophysin as an immunocytochemical marker of NE differentiation.

Topics: Animals; Animals, Newborn; Cell Differentiation; Epithelial Cells; Fluorescent Antibody Technique; Histocytochemistry; Humans; Immunoassay; Keratins; Lung; Lung Neoplasms; Membrane Proteins; Mice; Neurosecretory Systems; Rabbits; Synaptophysin

The histological diagnosis of malignant mesothelioma of the pleura, especially the distinction from peripheral adenocarcinoma of the lung, may be difficult. The immunohistochemical reports previously published on this subject show diverging results mainly because a variety of antibodies and staining techniques have been used by the different authors. To obtain comparable and reproducible results standard techniques and commercialized antibodies should be applied in routine pathology. In order to investigate the value of immunohistochemistry for the separation of the two entities formalin fixed and paraffin embedded blocks of 47 mesotheliomas and 22 adenocarcinomas were investigated with the PAP technique and commercially available antibodies to carcino-embryonic antigen (CEA), keratin, vimentin, epithelial membrane antigen (EMA), pregnancy specific antigen (SP1), S-100 protein and monoclonal antibody lu-5 (mAB lu-5). CEA positivity was found in all 22 adenocarcinomas examined, but only 2/47 (4%) of all mesotheliomas showed a positive result. SP1 was positive in 13/22 (59%) of the adenocarcinomas, whereas only 3/47 (6%) mesotheliomas were positive for this marker. No significant difference in the rate of positive cases in the adenocarcinoma and mesothelioma group could be found with the other above mentioned antigens. The results of our study indicate that especially CEA, but also SP1 are valuable markers in the diagnosis of malignant mesothelioma.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoembryonic Antigen; Diagnosis, Differential; Histocytochemistry; Humans; Immunochemistry; Keratins; Lung Neoplasms; Membrane Proteins; Mesothelioma; Mucin-1; Pleural Diseases; Pregnancy-Specific beta 1-Glycoproteins; Vimentin

Using a polyclonal antibody against human epidermal keratins and a monoclonal antibody against cytokeratins characteristic of simple epithelia, and the Avidin-Biotin system of immunohistochemistry, we have demonstrated cytokeratin expression in 46% and in 60% of small cell carcinomas of the lung at autopsy respectively. The latter gave a diffuse stronger reaction product than the polyclonal antibody. The results suggest that there is a cytokeratin rich and a cytokeratin poor type of small cell carcinoma. Neuron-specific enolase immunohistochemistry was positive in 60% of the cases. Coexpression with cytokeratin was seen in ten cases (30%). The expression of cytokeratin and neuron-specific enolase in small cell carcinomas strongly suggests that they are of an epithelial origin, but are capable of neuroendocrine differentiation.

Topics: Aged; Autopsy; Carcinoma, Small Cell; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Male; Middle Aged; Phosphopyruvate Hydratase; Retrospective Studies

The correct distinction between malignant mesothelioma of the pleura and adenocarcinoma of the lung has become increasingly complex, with a variety of histochemical, immunohistochemical, and ultrastructural studies to be performed on biopsy material. The reliability of immunohistochemical studies has been hampered by the use of polyclonal antisera to "carcinoembryonic antigen (CEA)" and keratin. Hybridoma technology now offers monoclonal antibodies (MAbs) in unlimited quantity and standardized quality to selective ranges of specific antigenic determinants. MAb B72.3, generated against a membrane-enriched fraction of human metastatic breast carcinoma, was used to distinguish malignant mesothelioma of the pleura from adenocarcinoma of the lung in tissue sections and was compared in terms of diagnostic utility with polyclonal anti-keratin and anti-CEA to make the same distinction. Reactivity with MAb B72.3 in at least 10% of tumor cells or more was noted in 19 of 22 adenocarcinomas of the lung (P greater than 0.0001), whereas none of the 20 cases of malignant mesothelioma demonstrated comparable reactivity. Furthermore, MAb B72.3 showed no reactivity with benign mesothelial proliferations. MAb B72.3 thus appears to be an appropriate diagnostic adjunct capable of discriminating between these malignancies.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Neoplasm; Carcinoembryonic Antigen; Diagnosis, Differential; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma; Pleural Neoplasms

Immunoperoxidase staining for human keratin proteins was performed cytologically on samples from 90 patients with malignant tumors, and histologically on samples from 164 patients with malignant tumors. At the cytological level, almost all tumor cells not only in squamous cell carcinoma but also in nonsquamous cell carcinoma were positive for keratin proteins, in contrast with the apparent abscence of keratin proteins in sarcoma. At the histological level, almost all neoplastic cells of squamous cell carcinoma were positive for keratin proteins, the same as at the cytological level. In contrast, among cases of nonsquamous cell carcinoma, the frequency of appearance of keratin proteins varied according to the organ; it tended to be low in tumors with relatively good prognosis, such as carcinomas in the digestive system or thyroid cancer, and to be high in tumor with poor prognosis, such as pulmonary cancer, gallbladder cancer and endometrial cancer. However, there was a marked difference between the frequency of appearance of keratin proteins at the cytological level and that at the histological level, particularly in the cases of gastric cancer.

Topics: Antibodies; Breast Neoplasms; Colonic Neoplasms; Female; Gallbladder Neoplasms; Histocytochemistry; Humans; Keratins; Lung Neoplasms; Neoplasms; Sarcoma; Stomach Neoplasms; Thyroid Neoplasms; Uterine Neoplasms

The phenotypic heterogeneity of epithelial cells during hematogenous metastasis from primary mammary carcinomas of BALB/cfC3H mice was examined. Antisera to keratins were used in immunofluorescence assays to identify epithelial and myoepithelial cells. Both cell types were located in frozen sections of not only the primary lesions but also pulmonary metastases. Tumor cells disseminating to the lungs were recovered from the bloodstream of tumor-bearing mice with the use of a quantitative density gradient centrifugation procedure. Carcinoma cells were detected in the circulation of 62% of mice at the time of assay, but all tumors examined had released cells into the circulation at some point in their history. Epithelial cells and myoepithelial cells were detected simultaneously in 21% of mice, whereas other mice had only epithelial cells (16%) or myoepithelial cells (25%) in their circulation at the time of assay. Single cells and multicellular emboli of each cell type were disseminated, together with similar numbers of heterogeneous emboli containing both epithelial and myoepithelial cells. The evidence shows that the presence of both mammary epithelial and myoepithelial cells in pulmonary metastases from mouse mammary carcinomas can be accounted for by their hematogenous delivery as differentiated cells.

Topics: Animals; Epithelium; Female; Keratins; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Neoplastic Cells, Circulating; Phenotype

To evaluate the usefulness of an immunohistologic approach to the differential diagnosis of mesothelioma and pulmonary adenocarcinoma, the authors studied paraffin-embedded, fixed tissue sections from 50 primary adenocarcinomas of the lung and 28 mesotheliomas of the pleura by using a panel of monoclonal antikeratin, antihuman milk fat globule (HMFG-2), anti-Leu M1, and monoclonal anticarcinoembryonic antigen (CEA) antibody; we also used a conventional heterologous anti-CEA antiserum with and without prior absorption with spleen powder to remove antibodies to nonspecific cross-reacting antigen (NCA). Keratin was present in both mesotheliomas and adenocarcinomas and did not help in distinguishing between these two neoplasms. HMFG-2 was detected in 48 (96%), and Leu M1 was positive in 47 (94%) of the adenocarcinomas, but not in any of the mesotheliomas. By using conventional rabbit antiserum, the authors detected CEA in the majority of adenocarcinomas (96%), but also in two cases of mesothelioma. When the anti-CEA antiserum was absorbed with NCA, the number of positively reacting adenocarcinomas decreased considerably to 76%; however, after this treatment, none of the mesotheliomas gave positive reactions. The monoclonal anti-CEA antibody was reactive in 36 of the adenocarcinomas (72%), but in none of the mesotheliomas. Our results indicate that, in addition to HMFG-2 and CEA, the expression of Leu M1 antigen by most primary pulmonary adenocarcinoma (94%) and its absence in mesothelioma could be used as a valuable marker for primary adenocarcinoma of the lung that involves the pleura and permits its differentiation from mesothelioma.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; Carcinoembryonic Antigen; Diagnosis, Differential; Humans; Keratins; Lung Neoplasms; Membrane Proteins; Mesothelioma; Mucin-1; Phenotype; Pleural Neoplasms

Paraffin sections of formalin-fixed tumor samples from 26 patients with neuroendocrine (Merkel cell) carcinoma of the skin (NECS) were studied immunohistochemically with three monoclonal antibodies to low molecular weight keratin (MAB-K) and with antibodies to leukocyte common antigen (LCA), neurofilament (NF), neuron-specific enolase (NSE), S100 protein (S100), and chromogranin (CGN), to investigate the relative diagnostic value of these antibodies. Samples from 20 lymphomas, 10 non-oat cell undifferentiated carcinomas, 10 oat cell carcinomas, and 10 melanomas served as controls. Keratin was found in 25 of the 26 NECS and in all undifferentiated and oat cell carcinomas. A ball-like immunostaining for keratins, resembling an inclusion body was seen only in cases of NECS and some carcinoids. Neurofilament, NSE, and CGN were expressed by fewer NECS than was keratin and all NECS were negative for LCA and S100. None of the lymphomas and melanomas contained detectable keratin, NF, NSE, or CGN. Only the lymphomas stained with LCA. Only the melanomas were S100-positive. It is concluded that keratin is the most useful single discriminating marker in the separation of neuroendocrine (Merkel cell) carcinoma of the skin from lymphoma, melanoma and, when the characteristic inclusion-like pattern is seen, from metastatic oat cell carcinoma.

Topics: Adult; Aged; Antibodies, Monoclonal; Carcinoid Tumor; Carcinoma; Carcinoma, Small Cell; Diagnosis, Differential; Female; Gastrointestinal Neoplasms; Histocytochemistry; Humans; Immunochemistry; Keratins; Lung Neoplasms; Lymphoma; Male; Melanoma; Microscopy, Electron; Middle Aged; Nerve Tissue Proteins; Skin Neoplasms

A clonal continuous cell line consisting of pre-differentiated lung epithelial cells was established from a Syrian golden hamster fetus on day 15 of gestation. Immunocytological techniques revealed cytoplasmic filaments positive to antikeratin antibody and positive to antivimentin antibody in these cells. Supplementation of the medium with epidermal growth factor, insulin, other types of hormones and vitamin A markedly enhanced cell growth and DNA synthesis. Simultaneously, the cell differentiation characterized by mucus-like glycoprotein production was greatly stimulated. Removal of vitamin A diminished cell growth and this differentiation. The cell system was found to be useful for studying the relationship between in vitro cell transformation and differentiation. Additionally, studies to define general chemical toxicities in this cell system by evaluating several parameters such as cell survival, sialic acid content and enzyme leakage into the medium are in progress.

Topics: Animals; Cell Differentiation; Cell Line; Cell Transformation, Neoplastic; Cricetinae; Cytoskeleton; DNA; Female; Fetus; Fluorescent Antibody Technique; Glucosamine; Keratins; Lung Neoplasms; Mesocricetus; Mucus; Vimentin; Vitamin A

The expression of vimentin in pulmonary carcinomas was studied in 285 cases of surgically resected lung cancer from our hospital files. Formalin fixed, paraffin-embedded sections were studied by immunoreactive staining techniques using two monoclonal antibodies against vimentin. Cases demonstrating vimentin positivity by the avidin-biotin-peroxidase method included 11 of 129 adenocarcinomas studied (8.5%), and 15 of 61 large cell carcinomas studied (24.6%). Vimentin expression was not seen in any of the 51 squamous cell carcinomas or 35 small cell carcinomas in our series. The positive cases of adenocarcinoma were in moderately and poorly differentiated cancers. Four of the eight giant cell carcinomas (50%) demonstrated vimentin expression. All cases that exhibited vimentin positivity were studied for cytokeratin expression. Coexpression of vimentin and cytokeratin was demonstrated not only within the same tumor but also within the same cells in some cases stained by double antibody technique, including both adenocarcinomas and large cell carcinomas. Similar immunoreactive methods were also applied to sections from human lung cancer transplants grown in the nude mouse. Of 28 tumors studied, four of 11 adenocarcinomas (36%) and all 4 large cell carcinomas demonstrated coexpression of vimentin and cytokeratin, while none of the five squamous cell carcinomas or eight small cell carcinomas expressed vimentin.

Topics: Adenocarcinoma; Animals; Antibodies, Monoclonal; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Histocytochemistry; Humans; Keratins; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Transplantation, Heterologous; Vimentin

Forty one cases of large cell anaplastic carcinoma of the lung (LCACL) were investigated by electron microscopy and immunoperoxidase studies for cytokeratin, enolase, and carcinoembryonic antigen. The results indicated that these neoplasias, grouped as an unique entity by ordinary histopathologic findings, may be further divided into five groups as follows: squamous, adenomatous, adenosquamous, neuroendocrine, and undifferentiated. The authors suggest that this subclassification may be useful in treatment orientation and in the prognostic evaluation of these neoplasias.

Topics: Carcinoembryonic Antigen; Carcinoma, Small Cell; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase

An enzyme-linked immunosorbent assay (ELISA) for anti-keratin antibodies was prepared by coating microplates with epidermal keratin purified from the stratum corneum from human foot. Naturally occurring auto-antibodies bind keratin via their f(ab')2 fragment. They were assayed in the serum from 65 healthy people. The serum titer increased significantly with aging. Auto-antibodies stained all the layers of a normal human epidermis whereas an immune serum, prepared by injecting a rabbit with pure human keratin proteins, stained more efficiently the outer layers of the human skin; pre-immune rabbit serum did not stain human skin at all. The lowest anti-keratin activities were observed in serum from patients with squamous cell lung carcinoma and with mesothelioma. The activity was lower in pleural fluid from patients with pleural mesothelioma than in pleural fluid from other types of cancer. This is possibly due to the fixation of autoantibodies onto the pleural tumor or on cell debris arising from the tumor.

Topics: Animals; Antibodies; Autoantibodies; Carcinoma, Squamous Cell; Enzyme-Linked Immunosorbent Assay; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Diseases; Lung Neoplasms; Mesothelioma; Pleural Effusion; Rabbits

The results of neuron-specific enolase (NSE) and keratin immunodetection in cytological specimens of sputum secured from 41 patients with lung cancer are presented. All 19 cases of small-cell carcinoma showed intense immunoreactivity for NSE. No such immunoreactivity was found in 21 of 22 cases of non-small-cell carcinoma. The single positive result was from a case of large-cell undifferentiated carcinoma. All 10 cases of squamous-cell carcinoma showed immunoreactivity for keratin. The 19 cases of small-cell carcinoma showed no such reactivity. Our findings indicate that immunostaining for NSE and keratin is a valuable aid when a definite diagnosis of small-cell carcinoma of the lung can not be made on the basis of conventional cytologic features.

Topics: Adenocarcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Clinical Enzyme Tests; Diagnosis, Differential; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Phosphopyruvate Hydratase; Sputum

A case of benign clear cell tumor of the lung was investigated immunohistochemically and histochemically. The tumor cells showed a packed proliferation pattern composed of two cell types. The one had polygonal, abundant eosinophilic cytoplasm, and the other had clear cytoplasm. These cells contained glycogen in the cytoplasm, detected by PAS and diastase treatment. Immunohistochemically, the tumor was strongly and diffusely stained by anti-vimentin antibody. And anti-NSE and S-100 protein antibodies were also positive in part. But anit keratin, Factor VIII, desmin, myoglobin and myosin antibodies were entirely negative. These results suggest that this tumor is a special neoplasia originating from neural tissue.

Topics: Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged; Myoglobin; Phosphopyruvate Hydratase; S100 Proteins; Vimentin

Forty-seven surgically resected small cell lung carcinomas (SCLC) were immunohistochemically studied by using antibodies to various neuroendocrine and epithelial markers. SCLC was shown to be subdivided into two categories, with and without the immunoreactive neuroendocrine markers aromatic L-amino acid decarboxylase, gastrin-releasing peptide, serotonin, chromogranin A and neurofilament protein. Neuron-specific enolase (NSE) and creatine kinase BB isoenzyme (CK-BB), which are also considered to be neuroendocrine markers, had a tendency to be widely distributed in the SCLC with a neuroendocrine marker, but the immunoreactivity for both NSE and CK-BB varied in the SCLC without neuroendocrine markers. Therefore they were not included in the classification. Epithelial markers keratin, involucrin and epithelial membrane antigen were frequently observed in the SCLC with neuroendocrine markers, but less so in the SCLC without neuroendocrine markers. The data are discussed briefly in relation to "classic and variant" forms of SCLC in vitro and to a recently proposed histological classification of SCLC.

Topics: Aromatic-L-Amino-Acid Decarboxylases; Carcinoma, Small Cell; Creatine Kinase; Gastrin-Releasing Peptide; Histocytochemistry; Humans; Immunochemistry; Isoenzymes; Keratins; Lung Neoplasms; Neurosecretory Systems; Peptides; Phosphopyruvate Hydratase; Protein Precursors

The intermediate filament protein (IFP) characteristics of a panel of lung cancer cell lines including adenocarcinoma (two cell lines) and small cell lung cancer (SCLC, three classic and three variant cell lines) were examined using one- and two-dimensional gel electrophoretic techniques, immunocytochemical techniques and immunoblotting assays. A panel of 28 monoclonal and polyclonal antibodies to the five different types of IFP were used. The results of our studies indicate that these human lung adenocarcinoma, classic SCLC and variant SCLC cell lines can be differentiated on the basis of their pattern of IFP. The main conclusions from this study can be summarized as follows. The two adenocarcinoma cell lines contain cytokeratins 7, 8, 18, and sometimes 19, next to vimentin intermediate filament (IF). The three classic-type SCLC cell lines contain only cytokeratin IFs but not vimentin IF or neurofilaments (NFs). Cytokeratin polypeptides 7, 8, 18 and 19 could be detected. All three variant-type SCLC cell lines do not contain detectable amounts of cytokeratins. In contrast, two out of three variant SCLC cell lines contain neurofilament proteins. All three variant-type SCLC cell lines contain vimentin IF. Using immunoblotting assays with monoclonal and polyclonal antibodies to defined NF proteins the presence of the 68 X 10(3) Mr and the 160 X 10(3) Mr NF polypeptide could be demonstrated in two variant SCLC cell lines. As patients with SCLC-variant phenotype have a poorer prognosis after cytotoxic therapy than patients with 'pure' SCLC, the use of antibodies to IFP in staining fresh lung tumours, especially anaplastic ones, may differentiate the two subtypes of SCLC. Such a distinction would have a major impact on therapy selections and may be of prognostic importance.

Topics: Antibodies, Monoclonal; Carcinoma, Small Cell; Cell Line; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Humans; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Vimentin

The occurrence and coexpression of the cytoskeletal proteins vimentin and cytokeratins were studied in malignant mesotheliomas and pulmonary carcinomas. For this purpose a double immunoenzyme staining with monoclonal antibodies was developed which made it possible to visualize vimentin and cytokeratins simultaneously within the same cell. A clear distinction between stromal cells (vimentin only) and tumour cells was also obtained. A total of 12 mesotheliomas (six mixed type and six epithelioid type) and 13 carcinomas (eight adenocarcinomas and five large cell undifferentiated carcinomas) were studied. The results revealed a clear difference between mesotheliomas and adenocarcinomas: 11 of 12 mesotheliomas showed coexpression of vimentin and cytokeratins in at least 50% of the tumour cells, while in seven of the eight adenocarcinomas none or only a few cells could be seen with this coexpression. In the undifferentiated large cell carcinomas three of five expressed both components, but in less than 25% of the cells. It is concluded that a reliable double immunoenzyme staining of vimentin and cytokeratins can be used as an additional means to distinguish malignant mesothelioma from pulmonary adenocarcinoma.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma; Peritoneal Neoplasms; Pleural Neoplasms; Staining and Labeling; Vimentin

Topics: Antigens, Neoplasm; Carcinoma, Squamous Cell; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Pleural Neoplasms

The expression of intermediate filament proteins in classic and variant-type small-cell lung carcinoma (SCLC) cell lines was studied using immunocytochemical techniques, two-dimensional gel electrophoresis and immunoblotting assays. Classic SCLC cell lines contain cytokeratin proteins but no neurofilaments. In contrast, variant cell lines do not contain detectable amounts of cytokeratins but partly express neurofilaments and vimentin. These results explain apparent discrepancies on the intermediate filament content of SCLC described in the recent literature. The application of antibodies to fresh biopsy specimens of SCLC may in the future allow the identification of the variant type of cells in clinical SCLC specimens and may have a major impact on therapeutic strategy and prognosis in these patients.

Topics: Carcinoma, Small Cell; Cell Line; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Humans; Immunosorbent Techniques; Intermediate Filament Proteins; Isoelectric Focusing; Keratins; Lung Neoplasms; Vimentin

Lung tumors of various types, fixed in 4% formaldehyde-1% glutaraldehyde, were stained for keratin proteins. The results were compared with previous ultrastructural evidence of intermediate filament bundles (IFBs), presumed to be keratin. Electron and light microscopic methods were largely complimentary and the results were in agreement in 79% of cases. Light microscopy was superior for demonstrating keratinizing foci containing numerous well-developed IFBs, whereas electron microscopy was superior when keratin filaments were sparsely distributed in cells throughout a tumor. Fetal and adult bronchial specimens were also studied. Normal adult bronchus, fixed in aldehydes, was unreactive but keratin was observed in similarly fixed bronchi that showed epidermoid metaplasia and/or intraepithelial carcinoma. Keratin was demonstrated in normal adult bronchi fixed in ethanol. Keratin was not observed in fetal lung until the 14th week of gestation, when it appeared in basal cells and a few columnar cells of the larger bronchi. Thereafter, keratin progressively appeared in the more distal branches. As specimens from gestations of less than 14 weeks were fixed in aldehydes, the apparent lack of immunoreactivity may have been artifactual. Nevertheless, keratin was demonstrable in aldehyde-fixed fetal bronchi at 16 and 23 weeks' gestation.

Topics: Adult; Carcinoma, Bronchogenic; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Cytoskeleton; Fetus; Histocytochemistry; Humans; Infant, Newborn; Keratins; Lung Neoplasms; Phenotype

Twenty-seven cases of surgically resected large cell carcinoma of the lung including nine cases of giant cell carcinoma were examined ultrastructurally and immunohistochemically. Ultrastructurally, of 18 large cell carcinomas other than giant cell carcinoma eight showed characteristic differentiation toward adenocarcinoma, four toward adenosquamous carcinoma, and one each toward squamous cell carcinoma and neuroendocrine cell carcinoma, but the remaining four were undifferentiated. Six of the nine giant cell carcinomas also showed features of adenocarcinoma, two showed features of squamous cell carcinoma, and one was undifferentiated carcinoma. Immunohistochemically, secretory component (SC) was observed in seven of 14 cases with features of adenocarcinoma and two of four cases with features of adenosquamous carcinoma. Carcinomas with only squamous cell differentiation did not stain for SC. Keratin staining was positive in five of the 14 with features of adenocarcinoma, three of the four cases with features of adenosquamous carcinoma and two of the three cases with features of squamous cell carcinoma. The numbers of tumor cells positive for keratin and/or SC were small. One carcinoma with neurosecretory type granules was stained positively for calcitonin. These findings indicate that many large cell carcinomas showed differentiation toward glandular cells and/or squamous cells, and some did not show any differentiation ultrastructurally or immunohistochemically, indicating that the majority of large cell carcinomas are poorly differentiated form of either adenocarcinomas or squamous cell carcinomas.

Topics: Adenocarcinoma; Calcitonin; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cytoplasmic Granules; Gastrin-Releasing Peptide; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Peptides; Secretory Component; Staining and Labeling

To elucidate the histogenesis of sclerosis haemangioma of the lung, we examined 7 cases with the immunoperoxidase method using antibodies against several useful marker antigens; secretory component (SC), cytokeratins, epithelial membrane antigen (EMA) for epithelial cells, factor VIII related antigens (factor VIII) for endothelial cells, vimentin and desmin for mesenchymal cells. The results were compared with those of histologically normal lung tissues. Both the characteristic round cells arranged in sheets, which are present predominantly in the solid area and are reported to be neoplastic, and the flattened cells lining blood lakes show positive staining for EMA only, with negative staining for the other marker antigens. These observations suggest that these cells are derived from epithelium rather than mesothelium or from endothelium, and are analogous to type I pneumocytes. This conclusion is supported by their immunohistochemical characteristics, in comparison with the localization patterns of the marker antigens in normal lung tissues. However, the lining epithelial cells of papillary projections in the papillary area and of ducts in the solid area stained for SC and cytokeratins as well as EMA, and their immunohistochemical characteristics are analogous to those of bronchiolar epithelial cells or type II pneumocytes in normal lung tissues.

Topics: Adult; Antibodies, Monoclonal; Antigens, Surface; Female; Hemangioma; Histocytochemistry; Humans; Immunochemistry; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Middle Aged; Secretory Component

A monoclonal antibody, 44-3A6, was raised against the human pulmonary adenocarcinoma cell line A549. This antibody recognizes a protein antigen at the cell surface, which is preserved after formalin fixation and paraffin embedding. Immunohistochemical staining of lung tissue with this antibody revealed diffuse immunoreactivity of type II pneumocytes. Bronchial epithelial cells were also focally immunoreactive. Immunostaining of various bronchopulmonary carcinomas demonstrated characteristic patterns of reactivity. All of the 42 adenocarcinomas and 18 carcinoids were strongly immunoreactive either diffusely or focally. The immunoreaction occurred at the cell membrane and/or in the cytoplasm. None of the 39 squamous cell carcinomas, 12 bronchioloalveolar carcinomas, and 30 small cell neuroendocrine carcinomas was immunostained. Ten intermediate cell neuroendocrine carcinomas and 8 well-differentiated neuroendocrine carcinomas were relatively weakly immunoreactive, while 7 and 2 of them were negative. Six adenosquamous carcinomas were focally positive in glandular and "basaloid" areas, whereas squamous areas were negative. Twenty-one large cell carcinomas were focally immunoreactive, while 6 were negative. It appears that MCA 44-3A6 is an effective marker for certain features of "glandular" differentiation, which may be present even in tumors lacking obvious glands, and that it may be useful for the differential diagnosis of various bronchopulmonary carcinomas.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoma; Diagnosis, Differential; Histocytochemistry; Humans; Keratins; Lung Neoplasms

Fifty-four human lung tumours have been immunostained with a large panel of monoclonal antibodies including reagents against cytokeratins, prekeratins, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA) and neural antigens. These results have been compared with the histological types of tumour using the current WHO classification scheme. The most striking finding of this study was the considerable overlap of antigenic profile between different histological types of tumour. This suggests that there may be a greater underlying similarity between different histological categories of lung tumour than has hitherto been assumed. Secondly it was evident that immunostaining highlighted areas of different morphology within many tumours emphasizing the heterogeneous differentiation patterns seen in lung cancers. The present study supports the viewpoint that lung tumours arise from a common stem cell and that these neoplasms represent a single tumour with a tendency to differentiate along one or more pathways.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Carcinoembryonic Antigen; Histocompatibility Antigens Class II; Histocytochemistry; HLA-DR Antigens; Humans; Immunochemistry; Keratins; Lung Neoplasms; Membrane Proteins; Mucin-1; Nerve Tissue; Protein Precursors

The intermediate filament cytoskeleton of epithelial, biphasic, and fibrous malignant pleural mesotheliomas was studied by immunohistochemistry and gel electrophoresis. The results were compared with data similarly obtained from lung adenocarcinomas. All mesotheliomas immunostained with various monoclonal and polyclonal antibodies against cytokeratins. By double immunofluorescence microscopy, coexpression of cytokeratins and vimentin was found in the fusiform cells of biphasic and fibrous mesotheliomas. As determined by two-dimensional gel electrophoresis, lung adenocarcinomas exclusively expressed Cytokeratins 7, 8, 18, and 19, and the same polypeptides were found in the fibrous mesotheliomas. These four cytokeratins were also found in the epithelial and biphasic mesotheliomas, most of which, however, also expressed, additional cytokeratins, such as the basic Polypeptide 5 and, in some cases, Cytokeratins 4, 6, 14, and 17. The results demonstrate the epithelial nature of all types of malignant mesotheliomas and thus justify their classification as carcinomas. When epithelial morphology is evident, the pattern of cytokeratin expression is usually more complex, as indicated by the synthesis, in addition to the "simple epithelial" pattern (7, 8, 18, and 19), of certain cytokeratin polypeptides which hitherto have been presumed to be typical of stratified epithelia. This cytokeratin complexity and the coexpression of vimentin and cytokeratins in certain forms of mesotheliomas indicate that these tumors are a clearly distinct and complex group of carcinomas. Their special cytoskeletal filament protein expression should prove useful in differentiating mesotheliomas from other carcinomas, particularly from adenocarcinomas growing in the lung.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Cytoskeleton; Electrophoresis; Humans; Immunochemistry; Intermediate Filaments; Keratins; Lung Neoplasms; Mesothelioma; Microscopy, Fluorescence; Pleural Neoplasms; Vimentin

Topics: Bronchi; Histocytochemistry; Humans; Immunologic Techniques; Keratins; Lung Neoplasms; Mucous Membrane

We have examined 18 primary malignant lung tumours categorized as either carcinosarcoma, blastoma or spindle-cell carcinoma according to accepted criteria. Two monoclonal antibodies to keratins, CAM 5.2 and LP 34, were used to determine whether the non-epithelial or spindle-cell components of each tumour showed evidence of keratin expression. By this means the epithelial nature of the five tumours classified as spindle-cell carcinomas was confirmed. In all four pulmonary blastomas and in five of nine carcinosarcomas, the sarcomatous elements failed to stain for keratin but in the remaining four carcinosarcomas there was focal staining. The histogenesis of these tumours is discussed and it is suggested that the sarcomatous component of a carcinosarcoma may be derived from malignant epithelial cells by a process of mesenchymal metaplasia with a switch in intermediate filament type. It remains uncertain whether blastomas are derived from both endoderm and mesoderm, or from either one of these tissues, with one component representing complete metaplastic transformation.

Topics: Aged; Antibodies, Monoclonal; Carcinoma; Carcinosarcoma; Cell Transformation, Neoplastic; Female; Humans; Intermediate Filaments; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasms, Germ Cell and Embryonal

Topics: Carcinoid Tumor; Electrophoresis, Polyacrylamide Gel; Female; Histocytochemistry; Humans; Intermediate Filaments; Keratins; Lung Neoplasms; Microscopy, Electron; Middle Aged

Topics: Aged; Carcinoma, Adenoid Cystic; Carcinoma, Small Cell; Female; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Middle Aged

Human lung tumor cell lines established from the major histological types of lung cancer were examined by immunofluorescent staining techniques for their patterns of intermediate filament (keratin, vimentin, and neurofilament triplet protein) expression. All cell lines examined, both small cell lung carcinoma (SCLC) and non-SCLC (squamous cell carcinoma, adenocarcinoma, large cell carcinoma, and mesothelioma) contained keratin, consistent with their epithelial derivation. These lung carcinoma cell lines also expressed vimentin, the characteristic intermediate filament of mesenchymal cells in vivo. In light of the proposed neuroectodermal origin of SCLC, cell lines were also studied for neurofilament expression. Two of four SCLC tumor cell lines, as well as non-SCLC cell lines, showed no reactivity with antibodies to neurofilament triplet protein. Two of the SCLC cell lines stained weakly with anti-neurofilament antibody. Examination of specific keratin patterns in human lung tumor cell lines by selective immunoprecipitation with keratin antiserum and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that small-sized keratin proteins (Mr 44,000 to 52,000) were present in cell lines derived from SCLC and non-SCLC types of lung cancer. Tumor cell lines exhibiting squamous differentiation by light microscopic criteria (i.e., intracellular keratin, intercellular bridging, "pearl" formation, and/or individual cell keratinization) also displayed a preponderance of intermediate-sized keratins (Mr 57,000 and 59,000) and exhibited another feature of terminal keratinocyte differentiation (cross-linked envelope formation). Mesothelioma cell lines had varying keratin profiles. The presence of keratin proteins in all SCLC cell lines examined argues against a neuroectodermal origin for these tumors and is consistent with the notion that these tumors arise from a common bronchial "stem cell," similar to that from which other types of bronchogenic carcinomas arise.

Topics: Animals; Carcinoma, Squamous Cell; Cell Line; Cell Transformation, Neoplastic; Cytoskeleton; Fluorescent Antibody Technique; Humans; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Mesothelioma; Mice; Microfilament Proteins; Molecular Weight; Protein Precursors; Vimentin

Immunohistological methods were used to investigate the presence of carcinoembryonic antigen, beta 1 pregnancy specific glycoprotein, beta subunit of human chorionic gonadotrophin, human placental lactogen, calcitonin, and keratin in formalin fixed tissue from 29 malignant mesotheliomas and 27 pulmonary adenocarcinomas. Malignant mesotheliomas were negative for tumour markers except for the beta subunit of human chorionic gonadotrophin and keratin, one and 13 cases respectively being positive for these. Pulmonary adenocarcinomas, however, were frequently positive for tumour markers--namely, carcinoembryonic antigen, beta 1 pregnancy specific glycoprotein, beta subunit of human chorionic gonadotrophin, human placental lactogen, calcitonin, and keratin. The presence of carcinoembryonic antigen and beta 1 pregnancy specific glycoprotein within an intrathoracic tumour is strong evidence against its being of mesothelial origin.

Topics: Adenocarcinoma; Calcitonin; Carcinoembryonic Antigen; Chorionic Gonadotropin; Diagnosis, Differential; Humans; Keratins; Lung Neoplasms; Mesothelioma; Placental Lactogen; Pregnancy-Specific beta 1-Glycoproteins

The authors investigated the expression of keratin, carcinoembryonic antigen (CEA), and an epithelial marker derived from milk fat globule membranes in 12 mesotheliomas and 100 diverse adenocarcinomas with immunohistochemical methods. The authors employed a monoclonal antibody to keratin designated as AE1, as well as the following commercially available antisera: rabbit anti-whole human keratin, rabbit anti-CEA, and a monoclonal antibody to an epithelial factor designated as MFG-2. Expression of keratin was found in all the mesotheliomas and adenocarcinomas with antibody AE1 as well as with the rabbit antiserum; CEA was detectable in 65% of the adenocarcinomas but two mesotheliomas also reacted weakly. With antibody MFG-2, positive results were obtained in 85% of the adenocarcinomas and in none of the mesotheliomas. All of 64 (100%) breast-, lung- and ovary-derived adenocarcinomas immunostained positively with antibody MFG-2. This is of particular significance because pulmonary and ovarian adenocarcinoma frequently may be indistinguishable clinically and histologically from epithelial mesothelioma. The authors conclude that antikeratin antibodies are not useful in the distinction of adenocarcinoma from mesothelioma. Because of its greater sensitivity and specificity, MFG-2 is superior to CEA in this differential diagnosis.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Breast Neoplasms; Carcinoembryonic Antigen; Diagnosis, Differential; Female; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Mesothelioma; Ovarian Neoplasms; Staining and Labeling

Immunohistochemical staining utilizing a peroxidase-antiperoxidase (PAP) technique for keratin, carcinoembryonic antigen (CEA), human chorionic gonadotropin (HCG) and alpha-fetoprotein (AFP) was performed on paraffin sections from 72 cases of lung cancer obtained at autopsy. Positive reaction was shown in 44% of the cases for keratin, 77% for CEA, and 58% for HCG. AFP was positive in only one case of large cell carcinoma. Keratin was positive in 100% of squamous cell carcinoma, 53% of adenocarcinoma, 15% of small cell carcinoma and 45% of large cell carcinoma. CEA showed positive staining in 90% of squamous cell carcinoma, 88% of adenocarcinoma, 58% of small cell carcinoma and 69% of large cell carcinoma. CEA was the most useful tumor marker for detection of all types of lung cancer. HCG was positive in 30% of squamous cell carcinoma, 100% of adenocarcinoma, 23% of small cell carcinoma and 56% of large cell carcinoma.

Topics: Adenocarcinoma; alpha-Fetoproteins; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Chorionic Gonadotropin; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms

Small cell bronchial carcinoma (SCC) xenografts with differing sensitivity to cyclophosphamide (CY) were investigated using a variety of techniques. Two xenografts (HX78 and HX88) were relatively sensitive to CY, one xenograft (HX72) was inherently resistant to CY and a fourth xenograft (HX78Cy) was a CY induced resistant subline of HX78 and was unstable when maintained without CY exposure. Conventional light microscopy, cytology and electron microscopy examination of the xenografts revealed appearances consistent with SCC. An antikeratin antibody demonstrated the presence of keratin in the inherent and the induced resistant xenografts (and in the unstable revertant) but not in the two sensitive xenografts; the presence of keratin suggested squamous differentiation. Monolayer culture morphology of the sensitive HX78 and the unstable revertant was anchorage independent with the cells forming floating aggregates; the induced resistant subline of this xenograft (HX78Cy) showed, by contrast, flattened, angular adherent cells. Beta HCG production was detected in the monolayer culture supernatant of sensitive HX78 cells; the level of production of beta HCG was increased in the induced resistant HX78Cy cells. Karyotype and flow cytometry studies were also performed. The morphological responses of small cell lung cancer to treatment are discussed.

Topics: Animals; Carcinoma, Small Cell; Cell Transformation, Neoplastic; Cells, Cultured; Chorionic Gonadotropin; Cyclophosphamide; Drug Resistance; Flow Cytometry; Humans; Karyotyping; Keratins; Lung Neoplasms; Mice; Neoplasm Transplantation; Transplantation, Heterologous

Paraffin sections from fifteen cases of malignant diffuse mesothelioma of the pleura and five cases of bronchial adenocarcinoma infiltrating the pleura were examined with an antiserum specific for factor VIII related antigen and with antisera against various epithelial markers: keratin, carcinoembryonic antigen (CEA), fat globule membrane antigen and secretory component. In all adenocarcinomas all the epithelial markers were present whereas the factor VIII related antigen was absent. The distribution of the fat globule membrane antigens, keratin, secretory component and factor VIII related antigen varied from one mesothelioma to another. The mesotheliomas were generally negative for CEA. The three mesotheliomas which were positive for CEA were also positive for alcian blue after hyaluronidase treatment. Amongst the markers used, CEA seems the most useful for the differential diagnosis between carcinoma and mesothelioma. However, the simultaneous detection of several markers allows the characterization of various phenotypes. Some of them are close to the phenotypes of true adenocarcinoma. A relation between a given phenotype and the biological behaviour of the tumour has still to be demonstrated.

Topics: Adenocarcinoma; Antigens; Antigens, Neoplasm; Carcinoembryonic Antigen; Factor VIII; Humans; Immune Sera; Keratins; Lung Neoplasms; Membrane Proteins; Mesothelioma; Mucin-1; Pleural Neoplasms; Secretory Component; von Willebrand Factor

Light-microscopic immunocytochemistry and electron microscopy demonstrated that adenocarcinomas (AC) and squamous cell (epidermoid) carcinomas (SCCs) of human lung contained keratin proteins in the form of tonofilament bundles. However, moderately differentiated (md) SCCs contained abundant keratin, whereas poorly differentiated (pd) SCCs and all ACs contained lesser amounts. Lung tumors with the diagnosis of AC or SCC, as defined by WHO criteria, were also analyzed by immunoprecipitation techniques for the presence of keratin proteins. Regardless of the degree of tumor differentiation, SCCs contained a 44 kd keratin which was lacking in ACs. Interestingly, normal bronchial epithelium also contained the same 44 kd keratin. In addition, as SCCs became more differentiated, they exhibited even greater differences in the profile of synthesized keratins. Specifically, the relative abundance of the intermediate-sized keratins (57 and 59 kd) was increased in the md SCCs. Although keratin protein patterns appear to be a valuable adjunct in distinguishing AC from SCC, their usefulness as a diagnostic tool will require survey of a larger number of poorly differentiated tumors.

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Bronchi; Carcinoma; Carcinoma, Squamous Cell; Epidermis; Histocytochemistry; Humans; Immunochemistry; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Microscopy, Electron; Precipitin Tests

Keratin profiles of exfoliated mesothelial and adenocarcinoma cells were determined using antisera to different molecular weight keratins (45, 46, 55, 63 kdaltons) and the immunoperoxidase technic. Most metastatic adenocarcinomas in effusions stained for low (45, 46 kdaltons) and intermediate (55 kdaltons) molecular weight keratins but were negative for 63 kdalton keratin. In contrast, most reactive and malignant mesothelial cells in effusions stained strongly for 63 kdalton keratin and keratins of lower molecular weight. This is the first report of high molecular weight (greater than 60 kdaltons) keratin in exfoliated cells of nonepidermal origin. Differences in staining for 63 kdalton keratin between mesothelial and adenocarcinoma cells may help to distinguish these cells in effusions.

Topics: Adenocarcinoma; Breast Neoplasms; Female; Gastrointestinal Neoplasms; Genital Neoplasms, Female; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma; Molecular Weight

Using formalin-fixed, paraffin-embedded tissue and commercial antisera, we evaluated the usefulness of immunohistochemical staining for carcinoembryonic antigen (CEA) and keratin in the diagnosis of malignant mesothelioma. All 18 adenocarcinomas of lung examined stained for CEA, usually strongly, while only eight of 22 mesotheliomas stained for CEA and the staining was generally weak. Staining for keratin was observed in 10 of 22 mesotheliomas and 12 of 18 adenocarcinomas; there were no differences in intensity of staining between the groups. We conclude that strong diffuse staining for CEA favors a diagnosis of carcinoma, and negative staining for CEA is against a diagnosis of carcinoma, but these are relative and not absolute criteria. We find that staining for keratin is of no use in distinguishing these types of tumors.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma

One hundred and twenty seven cases of lung tumors were studied by the immunoperoxidase technique for the presence of CEA and beta-HCG. Twenty-nine of these tumors were additionally stained for keratin and SP1, CEA and SP1 could be demonstrated in 80% of the studied cases, while beta-HCG was found in only 9%. SP1 revealed an almost identical staining pattern to CEA and keratin was found only in squamous cell carcinomas. The tissue positivity of none of these three markers correlated with tumor size, lymphnodal involvement or histological type.

Topics: Adenocarcinoma; Aged; Bronchial Neoplasms; Carcinoembryonic Antigen; Carcinoid Tumor; Carcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Chorionic Gonadotropin; Female; Histocytochemistry; Humans; Keratins; Lung Neoplasms; Male; Middle Aged; Pregnancy Proteins; Pregnancy-Specific beta 1-Glycoproteins

The various epithelial cells of the lower respiratory tract and the carcinomas derived from them differ markedly in their differentiation characteristics. Using immunofluorescence microscopy and two-dimensional gel electrophoresis of cytoskeletal proteins from microdissected tissues we have considered whether cytokeratin polypeptides can serve as markers of cell differentiation in epithelia from various parts of the human and bovine lower respiratory tract. In addition , we have compared these protein patterns with those found in the two commonest types of human lung carcinoma and in several cultured lung carcinoma cell lines. By immunofluorescence microscopy, broad spectrum antibodies to cytokeratins stain all epithelial cells of the respiratory tract, including basal, ciliated, goblet, and alveolar cells as well as all tumor cells of adenocarcinomas and squamous cell carcinomas. However, in contrast, selective cytokeratin antibodies reveal cell type-related differences. Basal cells of the bronchial epithelium react with antibodies raised against a specific epidermal keratin polypeptide but not with antibodies derived from cytokeratins characteristic of simple epithelia. When examined by two-dimensional gel electrophoresis, the alveolar cells of human lung show cytokeratin polypeptides typical of simple epithelia (nos. 7, 8, 18 and 19) whereas the bronchial epithelium expresses, in addition, basic cytokeratins (no. 5, small amounts of no. 6) as well as the acidic polypeptides nos. 15 and 17. Bovine alveolar cells also differ from cells of the tracheal epithelium by the absence of a basic cytokeratin polypeptide. All adenocarcinomas of the lung reveal a "simple-epithelium-type" cytokeratin pattern (nos. 7, 8, 18 and 19). In contrast, squamous cell carcinomas of the lung contain an unusual complexity of cytokeratins. We have consistently found polypeptides nos. 5, 6, 8, 13, 17, 18 and 19 and, in some cases, variable amounts of cytokeratins nos. 4, 14 and 15. Several established cell lines derived from human lung carcinomas (SK-LU-1, Calu -1, SK-MES-1 and A-549) show a uniform pattern of cytokeratin polypeptides (nos. 7, 8, 18 and 19), similar to that found in adenocarcinomas. In addition, vimentin filaments are produced in all the cell lines examined, except for SK-LU-1.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: Adenocarcinoma; Animals; Carcinoma, Squamous Cell; Cattle; Cell Line; Cell Transformation, Neoplastic; Diagnosis, Differential; Humans; Keratins; Lung; Lung Neoplasms

A panel of established human pulmonary cancer cell lines, representing the major histopathologic groups according to the World Health Organization (WHO) classification (WHO 1, squamous cell carcinoma; WHO 2, small cell carcinoma; WHO 3, adenocarcinoma; WHO 4, large cell carcinoma) were examined for their expression of various types of intermediate filaments in order to determine their phenotypic differences and to attempt to disclose their histogenetic origin. The cells were investigated with antibodies specific for cytokeratin, vimentin, and neurofilament polypeptides with both immunofluorescence microscopy and immunoblotting techniques. Squamous cell carcinoma and adenocarcinomas expressed cytokeratin in accordance with the epithelial nature of these tumors but not neurofilament polypeptides. Small cell carcinomas, on the other hand, were positive for neurofilaments but negative for keratin. In contrast to small cell carcinoma, adenocarcinoma, and squamous cell carcinoma, one cell line derived from large cell carcinoma appeared to express both neurofilaments and keratin. All cell lines studied also contained variable amounts of vimentin, a phenotypic characteristic obtained by many cells under in vitro conditions. The results demonstrate, in accordance with our earlier observations in vivo, a distinctly divergent expression of intermediate filament proteins in different types of lung cancers. The persistence of this phenotypic heterogeneity in vitro consolidates the use of cell cultures as useful models to study the biologic behavior and interrelationships of lung cancers. Based on the present studies, and taking into account the occurrence of mixed forms of lung cancers, we present a hypothetical scheme of the histogenetic derivation of different types of lung cancers.

Topics: Adenocarcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line; Female; Fluorescent Antibody Technique; Humans; Immunochemistry; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Male; Microscopy, Fluorescence; Neoplasm Proteins; Phenotype; Vimentin

Three monoclonal antibodies, one directed against cytokeratin (clone 80) and two directed against neurofilament (clones 2F11 and 3G6), were used in the study of a series of 77 lung carcinomas by immunohistochemical staining. The anti-cytokeratin antibody, a very broadly reacting antibody directed against an antigenic determinant common to a great number of cytokeratins, was applicable on frozen sections. The two anti-neurofilament antibodies, directed against the 70 kD protein (clone 2F11) and the 160 kD and 200 kD proteins (clone 3G6) of neurofilament, were applicable on both frozen sections and paraffin sections. The staining results on the lung carcinomas indicate that all types of tumors studied, including small-cell anaplastic carcinoma, are markedly positive for cytokeratin. Frozen sections of five and formalin-fixed and paraffin-embedded sections of six other small-cell anaplastic carcinomas were negative with both anti-neurofilament monoclonal antibodies. One poorly differentiated squamous cell carcinoma positive with anti-neurofilament clone 2F11 but negative with clone 3G6. This distribution of cytoskeletal proteins demonstrates the epithelial differentiation of all types of lung carcinomas. Neuroendocrine differentiation of lung carcinomas as found in the small-cell anaplastic types does not result in expression of neurofilament proteins.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Antibodies, Monoclonal; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Histocytochemistry; Humans; Intermediate Filament Proteins; Keratins; Lung Neoplasms; Neoplasm Proteins; Neurofilament Proteins

A detailed ultrastructural study was made of seven cases of bronchiolo-alveolar carcinoma, and the findings were correlated with histochemical and immunohistochemical data. By electron microscopic examination all seven tumors displayed glandular differentiation, manifested by the presence of microvilli and intercellular junctions, with or without mucin production. Variable proportions of tumor cells retained ultrastructural characteristics of alveolar type II cells and Clara cells. In addition, some tumor cells revealed desmosomes and tonofilaments consistent with squamous differentiation. Immunohistochemical evaluation was carried out using a peroxidase-antiperoxidase technique and specific antibodies against surfactant high molecular weight glycoproteins, keratin proteins, IgA + secretory piece, carcinoembryonic antigen (CEA), human chorionic gonadotropin (HCG), and alpha-fetoprotein (AFP). Four tumors with type II cell-like differentiation stained with anti-surfactant glycoprotein sera. All seven tumors stained focally with anti-keratin and IgA + anti-secretory piece antibodies, and diffusely with CEA. These tumors failed to stain with antisera against HCG and AFP. It is concluded that bronciolo-alveolar carcinomas are primarily composed of cells with alveolar and bronchiolar cell differentiation. Adequate criteria were established for ultrastructural identification of tumor cells with differentiation to type II alveolar cell or Clara cell. Moreover, the findings of this study indicate that the surfactant glycoprotein marker, when present in a given tumor either diffusely or focally, is diagnostic of bronchiolo-alveolar carcinoma.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; alpha-Fetoproteins; Carcinoembryonic Antigen; Cell Differentiation; Chorionic Gonadotropin; Glycoproteins; Histocytochemistry; Humans; Immunoenzyme Techniques; Immunoglobulin A; Keratins; Lung Neoplasms; Male; Middle Aged; Proteolipids; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants; Secretory Component

Cigarette smoking is known to be an important etiologic factor in several lung diseases; however, the number of smokers who develop these diseases represents a small segment of the smoking population. It is possible that evidence of inhalation-induced injury to bronchial epithelial cells of smokers will be reflected in the proteinaceous products of these cells, thereby identifying a high-risk subgroup. We have tested this hypothesis by analysis of 2 proteins, free secretory component (FSC) and the keratins, in lavage fluids obtained from 4 groups of subjects: 30 normal nonsmokers, 15 asymptomatic smokers, 22 symptomatic smokers, and 40 carcinoma patients. Among symptomatic smokers, FSC relative to total protein (FSC/TP) was depressed compared with that in nonsmokers and asymptomatic smokers. The keratins were detected only in symptomatic smokers and correlated with pack/years of smoking history (p = 0.017). Carcinoma patients had depressed FSC/TP and detectable keratin (33 of 38 patients studied). Lung sections from carcinoma patients studied immunohistochemically revealed an apparent inverse relationship between tissue FSC and keratins. This inverse relationship was borne out by analysis of these proteins in the lavage fluid of cancer patients (r = -0.4, p = 0.04). Thus, in cancer patients, immunohistochemical evidence of airway injury correlates with bronchial lavage levels of mucosal epithelial cell proteins. It is possible that smokers with altered levels of these proteins may be the ones at increased risk of smoking-associated lung disease.

Topics: Adolescent; Adult; Aged; Bronchi; Carcinoma; Enzyme-Linked Immunosorbent Assay; Epithelium; Glycoproteins; Humans; Keratins; Lung Neoplasms; Middle Aged; Smoking; Therapeutic Irrigation

Immunoperoxidase staining for human keratin proteins (hKP) was performed cytochemically in samples from 79 cancer patients, and histochemically in samples from 134 cancer patients. Immunohistochemically, hKP was present in almost all patients with squamous cell carcinoma (lung), transitional cell carcinoma (urinary bladder), adenocarcinoma (lung, stomach, breast, ovary), bronchiole-alveolar carcinoma (lung), and large cell carcinoma (lung). It was detected in 40% of the patients with small cell carcinoma (lung). Histochemically, hKP was present in patients with squamous cell carcinoma (lung, uterine body), adenocarcinoma (lung, stomach, colon, gall bladder, thyroid, uterine body), adenosquamous cell carcinoma (gall bladder, uterine body), Signet-ring cell carcinoma (stomach), clear cell carcinoma (uterine body) and undifferentiated carcinoma (uterine body). However, it was not detected in patients with brochiole ++-alveola ++ carcinoma (lung) and mucinous carcinoma (gall bladder).

Topics: Breast Neoplasms; Colonic Neoplasms; Female; Histocytochemistry; Humans; Keratins; Lung Neoplasms; Male; Neoplasms; Stomach Neoplasms; Urinary Bladder Neoplasms; Uterine Neoplasms

Three systems--the indirect, peroxidase-antiperoxidase (PAP), and avidin-biotin peroxidase complex (ABC) detection system--were compared in quantitative ELISA titration and inhibition assays and tissue labeling with keratin antisera. The ABC system was found to be the most sensitive of the three in ELISA titration assays, being approximately 4-fold more sensitive than the indirect method and 2-fold more sensitive than the PAP method. In addition, the ABC method demonstrated increased sensitivity when compared on tissue sections. Furthermore, the secondary and tertiary components of the PAP and ABC systems were titrated and optimal concentration ranges determined for use in ELISA inhibition assays. In ELISA inhibition assays the broadest usable inhibitor range and maximal sensitivity were obtained using the ABC system as compared with indirect and PAP detection systems. Finally, the effects of varying microtiter plate-coating concentrations on range and sensitivity were examined, and possible explanations are discussed including the possibility of surface-bound antigen being a competitor for the antibody of the solution phase antibody-antigen complex.

Topics: Antibodies; Avidin; Biotin; Carcinoma, Squamous Cell; Chemical Phenomena; Chemistry; Enzyme-Linked Immunosorbent Assay; Humans; Immunoenzyme Techniques; Isoenzymes; Keratins; Lung Neoplasms; Peroxidase; Peroxidases

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Diagnosis, Differential; Histocytochemistry; Humans; Immunochemistry; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma

Fifty-two primary carcinomas of the lung were studied by electron microscopy and by an immunoperoxidase method, using an anti-human keratin antiserum. The results were compared with light microscopic observations. One-third of the carcinomas of the lung showed ultrastructural evidence of both glandular and squamous differentiation. The group of small cell carcinomas was found to be particularly heterogenous ultrastructurally with only three out of eight tumors showing neurosecretory-type granules. Indirect immunoperoxidase staining revealed presence of a keratin-type protein in the vast majority of carcinomas, including foci of small cell carcinomas. Our studies emphasize the heterogeneity and frequent intermixing of the four major categories of lung carcinomas: squamous cell carcinomas, adenocarcinomas, small cell carcinomas, and large cell carcinomas. It is suggested that all these tumors might be derived from pluripotential "reserve" bronchial or bronchioalveolar cells. The segregation of small cell carcinomas from other groups continues to be justified on pragmatic grounds because these carcinomas constitute a group of predominantly small fast-replicating cells amenable to chemotherapy.

Topics: Adenocarcinoma; Carcinoma; Histocytochemistry; Humans; Immune Sera; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Male; Microscopy; Middle Aged

In this immunohistochemical study, antiserums to different molecular weight keratin proteins (45kd, 46kd, 55kd, and 63kd) were utilized to determine the profiles of keratin proteins present in a variety of pulmonary neoplasms. Different histologic types of lung carcinoma exhibited different patterns of keratin staining. Squamous cell carcinomas stained strongly for 45K, 46K, and 55K keratin, with staining for 63K restricted to areas or individual cells with cytoplasmic keratinization. Adenocarcinomas showed variable, generally weak staining for 45K, 46K, and 55K keratin and were uniformly negative for 63K keratin both in frozen and paraffin sections. Mesotheliomas and reactive mesothelial cells, by contrast, stained positively for 63K keratin in addition to keratins of lower molecular weights. Differences in staining for 63K keratin between mesothelioma and adenocarcinoma may have diagnostic application. Moreover, individual cytokeratins may serve as markers of tumor differentiation and provide information as to the origin of neoplastic cells.

Topics: Adenocarcinoma; Carcinoma, Squamous Cell; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma; Molecular Weight

The purpose of the present study was to compare the relative merits of cold acetone and buffered formalin as fixatives for the detection of carcinoembryonic antigen and keratin in permanently embedded tissues using a peroxidase-antiperoxidase (PAP) immunohistochemical procedure. The effect of treatment with the proteolytic enzyme pronase also was examined in the formalin-fixed tissues. Cold acetone was found to be superior to formalin for the preservation of CEA and keratin antigenic activities in a variety of benign and malignant tissues. Pronase treatment markedly increased the staining intensities of both antigens in formalin-fixed tissues. For many tissues, however, superior results were obtained using the cold acetone method, and this technic is recommended for the optimum retention of antigenic activity in permanently embedded tissues.

Topics: Acetone; Adenocarcinoma; Carcinoembryonic Antigen; Formaldehyde; Histocytochemistry; Histological Techniques; Humans; Immunochemistry; Keratins; Lung Neoplasms; Pronase; Staining and Labeling

Immunohistochemical localisation of keratin was assessed on 45 diagnostic specimens of small cell carcinoma of the lung in patients who subsequently received combination chemotherapy. Nine out of 45 (20%) contained keratin immunoreactive cells. Six of these achieved a complete response to treatment compared to 12 of the tumours which did not show positive staining for keratin. For 2 patients the tumours were shown to contain nests of keratin immunoreactive cells. Both of these are alive and free of disease more than 5 yr after the initial diagnosis. The results indicate that the presence of keratin immunoreactive cells may not directly equate with squamous differentiation and therefore not constitute an adverse prognostic factor in terms of response to chemotherapy.

Topics: Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Small Cell; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Prognosis

Human fetal lung tissue was cultured on dead, sterile pigskin as a substrate and treated with 3-methylcholanthrene. Several cell lines were obtained. One of the lines produced a tumor in nude mouse and showed anchorage-independent growth in soft agar. The tumor cells were stained positive for keratin, indicating their epithelial origin.

Topics: Antibodies, Neoplasm; Cell Transformation, Neoplastic; Cells, Cultured; Female; Fetus; Humans; Keratins; Lung Neoplasms; Methylcholanthrene

Antisera against total keratin extracts of human callus have been used to identify keratins in lung tumours of different histological type. Forty-three were classified by the WHO scheme. Keratin immunoreactive cells were identified in all 8 epidermoid carcinomas; 6 out of 12 large cell carcinomas; 2 out of 6 adenocarcinomas; 2 out of 15 small cell carcinomas and in the only muco-epidermoid carcinoma. These cases demonstrate the heterogeneity of phenotypic expression in lung tumours not recognisable without the use of immunohistochemical techniques.

Topics: Adenocarcinoma; Bronchial Neoplasms; Carcinoma; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Humans; Immune Sera; Keratins; Lung Neoplasms; Phenotype

The distribution of intracellular keratins was investigated in normal bronchial epithelium and in several morphologically distinct forms of respiratory tract carcinomas. This study was performed with two different experimentally produced antisera against normal human stratum corneum keratin and against keratin protein of MW 67,000 dalton, using indirect immunofluorescence and immunoperoxidase methods on tissue sections and cell suspensions. In normal bronchial epithelium, the basal cells were strongly labelled by both antisera. The ciliated columnar cells appeared devoid of cytokeratins in tissue sections but were strongly labelled with both antisera in cell suspensions. The goblet cells remained negative in every case. In squamous metaplasia of the bronchus, all epithelial cells were unevenly stained with both antisera. Among tumours, only the squamous cell carcinomas were strongly labelled by both antisera. Primary lung adenocarcinoma appeared weakly positive, whereas metastatic lung carcinomas, undifferentiated lung carcinomas, oat cell tumours, carcinoid tumours were negative. The immunocytochemical determination of keratins appeared to be of value in the study of normal and abnormal epithelial differentiation, in the diagnosis of poorly differentiated carcinomas and in their distinction from metastatic tumours of the lung.

Topics: Adenocarcinoma; Bronchi; Bronchial Neoplasms; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Epithelium; Humans; Keratins; Lung Neoplasms; Metaplasia

The distribution of keratin proteins and carcinoembryonic antigen (CEA) in 20 diffuse pleural malignant mesotheliomas and 20 adenocarcinomas of the lung was determined with the use of an indirect immunoperoxidase method. Keratin proteins were identified in all of the mesotheliomas, with strong staining observed in 17 of the cases. Tumor cells of various histologic types (tubular, papillary, solid, and spindle) revealed staining for keratin proteins. A variety of staining patterns were observed, but the homogeneous pattern predominated, in either a diffuse (16 cases) or focal form (4 cases). CEA was usually absent (11 cases), but weak or equivocal staining was also observed (8 cases), and 1 case uniquely exhibited moderate staining for CEA. In contrast, adenocarcinomas of the lung usually stained weakly or negatively (18 cases) for keratin proteins and exhibited a predominantly peripheral staining pattern. All cases, however, stained strongly or moderately for CEA. The profile of strong keratin staining and weak or absent CEA staining appears characteristic of mesotheliomas and may be diagnostically useful in defining the epithelial element of these neoplasms and in distinguishing them from adenocarcinomas.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Cytoplasm; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma; Pleural Neoplasms; Pneumothorax

An immunoperoxidase technique employing antibody to prekeratin was used to study distribution and pattern of staining of prekeratin filaments in cytological smears obtained from 42 specimens of pleural and peritoneal effusions (27 benign, 15 malignant). The smears were either air-dried or ethanol-fixed. Both benign and malignant mesothelial cells showed distinctive peripheral or perinuclear staining patterns which differed from the characteristic arborizing pattern in adenocarcinoma cells. The ultrastructure of these 2 cell types studied in 27 body fluids (12 benign, 15 malignant) and in 13 malignant tumors (3 mesotheliomas, 10 adenocarcinomas) showed a distinctive localizaton of intermediate filaments which corresponded to and could explain the pattern of staining obtained using the immunoperoxidase technique. The immunohistochemical and ultrastructural findings appeared characteristic for benign and malignant mesothelial cells as well as for adenocarcinoma cells, and could be used as markers to differentiate mesothelial tumors and reactive mesothelial cells from adenocarcinomas.

Topics: Adenocarcinoma; Ascitic Fluid; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Lung Neoplasms; Mesothelioma; Pleural Effusion; Protein Precursors

A continuous cell line, U-1752, was established from a lung tumor originally diagnosed as a small cell carcinoma. The cell line has been in continuous in vitro passage for 29 months. The epithelial, rather than small cell nature of the U-1752 cells was demonstrated by the presence of desmosomes, prominent tonofilament bundles, by the reactivity with an anti-keratin antiserum and by the expression of cell surface receptors for epidermal growth factor (EGF). The U-1752 cells grow as monolayer cultures and have a population doubling time of around 36 hours at optimal growth in 20% calf serum. The most important neoplastic features of U-1752 were its aneuploidy, its capacity for colony formation in agarose and its tumorigenic potential subcutaneously in nude mice.

Topics: Adult; Animals; Carcinoma, Squamous Cell; Cell Line; Chromosomes; Hormones; Humans; Immune Sera; Keratins; Lung Neoplasms; Male; Mice; Neoplasm Transplantation

Topics: Animals; Carcinoma; Goats; Keratins; Lung Neoplasms; Metaplasia; Microscopy; Mucins; Retrospective Studies