bromochloroacetic-acid and Head-and-Neck-Neoplasms

In the past decade, several emerging bone and soft tissue neoplasms of the head and neck region have been described in the literature, including GLI1-altered mesenchymal tumors, (intraosseous) rhabdomyosarcoma with TFCP2 fusion, and adamantinoma-like Ewing sarcoma. This review provides a summary of the clinical features, histologic characteristics, immunoprofile, key diagnostic features, and differential diagnoses of these emerging entities. Notably, all three entities show epithelioid morphology and cytokeratin immunopositivity, highlighting the need to consider these mesenchymal neoplasms in the differential diagnoses of cytokeratin-positive epithelioid tumors in the head and neck region. Appropriate workups including detection of the characteristic molecular alterations are essential for the correct diagnosis.

Topics: Biomarkers, Tumor; Bone Neoplasms; DNA-Binding Proteins; Head and Neck Neoplasms; Humans; Keratins; Los Angeles; Sarcoma, Ewing; Soft Tissue Neoplasms; Transcription Factors

Atypical Fibroxanthoma (AFX) is a rare cutaneous neoplasm arising from myofibroblast or fibroblast-like cells that predominantly affects the head and neck region. It commonly mimics more invasive neoplasms and is a diagnostic challenge to clinicians. The aim of this study was to develop a better understanding of AFX, focusing on recent developments in diagnosis and management.. A retrospective case series and review of recent literature were carried out.. Over a 17-year period, seven cases were identified (six male, mean age at presentation was 75.9 years). Two patients underwent complete excision and five patients had curettage and cauterisation. Two patients developed local recurrence but none demonstrated signs suggestive of metastatic spread. Histologically all seven lesions displayed a spindle cell pattern. Where performed, immunohistochemical staining was positive for Vimentin, CD10, CD68 and actin, and negative for CAM 5.2, CD34, Melan-A, S100 protein, HMB45, Cytokeratin A1/A3.. Our patient demographics, histopathology and immunohistochemistry are comparable to previous studies. Although advances have been made in immunohistochemical analysis, we are yet to discover a specific diagnostic immunostain for AFX. Clinical findings should therefore be correlated with histology and a panel of immunohistochemical stains should be used. Given the potential for recurrence or metastases, Moh's Micrographic Surgery with regular follow-up may be the preferred management.

Topics: Actins; Aged; Aged, 80 and over; Antigens, CD; Antigens, CD34; Antigens, Differentiation, Myelomonocytic; Biomarkers; Cautery; Cohort Studies; Curettage; Female; Head and Neck Neoplasms; Humans; Keratins; Male; MART-1 Antigen; Middle Aged; Mohs Surgery; Neoplasm Recurrence, Local; Neoplasms, Connective Tissue; Neprilysin; Retrospective Studies; S100 Proteins; Scalp; Skin Neoplasms; Vimentin

Topics: Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cell Proliferation; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Melanoma; Nose Neoplasms; S100 Proteins; Skin Neoplasms; Squamous Cell Carcinoma of Head and Neck

This article summarizes research using cells derived from epidermis of the miniature pigs for use as a cell therapy for skin repair and as a model for squamous carcinoma of the head and neck. Stem cells are an important "tool" for biomedical research. Adult stem cells are defined functionally, as cells that have the capacity to self-renew as well as the ability to generate differentiated cells. They are present in defined tissue microenvironments called niches. Asymmetric mitosis allows them to produce one daughter cell with the properties of stem cells (self-renewal) and a second cell with characteristics of progenitor cells, or transit amplifying cells, which proliferate quickly but with a limited number of mitotic divisions. Porcine epidermal stem cells, located in the bulge region of the outer root sheath of hair follicles, migrate in vitro from hair sheaths and because they are resistant to anoikis (detachment induced apoptosis), survive in non-adhesive conditions to form spheroids. These cells express keratins, galectin-1 and their nuclei are rich in DeltaNp63alpha. Interestingly, the multiple phenotype analysis of the human tumor cells in squamous carcinoma of head and neck revealed similarities with epidermal stem cells. These cancer stem cells are usually located on the periphery of the tumor where the invasive front of the tumor responsible for its aggressive behavior is located. In contrast, extensive expression of markers of terminal differentiation such as expression of glycoligands reactive for the endogenous lectin, galectin-3, indicates better tumor prognosis.

Topics: Animals; Cell Survival; Epidermal Cells; Galectin 1; Hair Follicle; Head and Neck Neoplasms; Humans; Keratins; Mitosis; Stem Cell Transplantation; Stem Cells; Swine; Wound Healing

Merkel cell carcinoma (MCC) is an uncommon primarily dermal malignancy of relatively aggressive biologic course. Several presentations in the mucosa of the head and neck region have been reported in the literature, and 3 such patients have recently been seen at our institution. We review this recent experience and present the first reported primary lingual MCC in a 57-year-old caucasian man. We provide a review of oral mucosal MCC and guidelines for histopathologic and immunohistochemical diagnosis. Merkel cell carcinoma should be included in the differential diagnosis of head and neck mucosal lesions, especially if the tumor is submucosal, and MCC may involve the tongue. Mucosal MCC is aggressive, and there is a high risk for local recurrence and regional and distant metastasis. Fulminating courses are often seen. We discuss our treatment policies based on the current literature.

Topics: Aged, 80 and over; Biomarkers; Carcinoma, Merkel Cell; Chromogranin A; Chromogranins; Fatal Outcome; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratin-20; Keratins; Lip Neoplasms; Male; Middle Aged; Mouth Mucosa; Phosphopyruvate Hydratase; Synaptophysin; Tongue Neoplasms

Topics: Adenocarcinoma; Biomarkers, Tumor; Brain Neoplasms; Breast Neoplasms; Carcinoma, Hepatocellular; Carcinoma, Neuroendocrine; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Differentiation; Cytogenetic Analysis; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Liver Neoplasms; Lymphatic Metastasis; Magnetic Resonance Imaging; Mesothelioma; Neoplasm Metastasis; Neoplasms, Unknown Primary; Peritoneal Neoplasms; Positron-Emission Tomography; Prognosis; Rhabdomyosarcoma; Tomography, X-Ray Computed; Urinary Bladder Neoplasms

The increasing interest in the so-called sentinel node concept, which has recently been adapted to squamous cell carcinomas of the upper aerodigestive tract, can be explained by encouraging results in other tumor entities. Although the publications on this topic do not yet allow a final judgment on the significance of sentinel lymphadenectomy in head and neck squamous cell carcinoma, early results emphasize the importance of this new diagnostic and therapeutic concept. The basic prerequisite is a detailed knowledge of the existing method-specific limitations in this anatomic region. Critical and careful evaluation of the sentinel node concept is mandatory prior to its application to other tumor entities. Sentinel lymphadenectomy for head and neck cancer may prove helpful if the indications for its use are clearly defined.

Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lymph Nodes; Lymphatic Metastasis; Radionuclide Imaging; Sentinel Lymph Node Biopsy; Tonsillar Neoplasms

An extremely rare hamartomatous lesion of the juxtaoral organ of Chievitz (JOOC) in a 63 year old man is reported. The tumour appeared as a large mass in the infratemporal fossa with associated mandibular bone resorption; histologically, it was well encapsulated and composed of numerous tangled masses of benign squamous epithelial nests and mature fibrofatty tissue. There were no histological features suggestive of neoplastic transformation. A literature survey confirmed that this is the first adult case of JOOC presenting clinically as an extraoral tumour.

Topics: Biomarkers; Cheek; Hamartoma; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Magnetic Resonance Imaging; Male; Middle Aged; Tomography, X-Ray Computed

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

The malignant rhabdoid tumor (MRT) has been a controversial lesion since its seminal description. There is no consensus as to whether it represents a distinctive clinicopathological entity or, alternatively, a phenotypic pattern that is potentially common to several disparate neoplasms. MRT of the kidney is a childhood tumor that is associated with uniformly aggressive behavior, but it shows a wide spectrum of histologic, immunophenotypic, and cytogenetic findings. Malignant extrarenal rhabdoid tumors (MERTs) have been observed in pure form over a broader range of patient ages and anatomic locations, but they show substantial morphological and biological homology with renal MRT. Lastly, "composite" extrarenal rhabdoid tumors (CERTs)--in which recognizable "parent" neoplasms are admixed with MERTs--also have been recognized in several topographic sites. In aggregate, these observations suggest that "rhabdoid tumors" are a heterogeneous group of lesions with dissimilar lineages of differentiation. Particularly in CERTs, it is likely that the rhabdoid phenotype represents a common end point of clonal evolution in tumors of clearly different origins. Despite these caveats, the authors do support retention of the diagnosis of "rhabdoid tumor," because the affiliated morphological pattern is uniformly attended by aggressive biological behavior despite potential dissimilarities at a subcellular level.

Topics: Abdominal Neoplasms; Central Nervous System Neoplasms; Child, Preschool; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Infant; Infant, Newborn; Keratins; Kidney Neoplasms; Male; Mucin-1; Rhabdoid Tumor

Six cases of squamous cell carcinoma arising in the head and neck of patients infected with the human immunodeficiency virus are described. This article reports the first two cases of primary intraosseous squamous cell carcinoma associated with infection with human immunodeficiency virus. Clinical presentation, results of imaging studies, histologic characteristics, therapies applied, and the clinical follow-up are described in detail for each of the six cases. These data are evaluated through a review of the current literature.

Topics: Acquired Immunodeficiency Syndrome; Adult; Biomarkers, Tumor; Carcinoma, Squamous Cell; CD4-CD8 Ratio; Female; Head and Neck Neoplasms; HIV Infections; Humans; Immunocompromised Host; Keratins; Male; Mandibular Neoplasms; Middle Aged; Mouth Neoplasms

Immunocytochemical analysis in fine-needle aspirates were performer in 46 cases of head and neck tumors. Immunocytochemical studies helped to establish the differential diagnosis of all diagnostically difficult cases. The application of 3 monoclonal antibodies against keratin, vimentin and LCA permitted to differentiate basic types of malignant tumors.

Topics: Antibodies, Monoclonal; Desmin; Diagnosis, Differential; Glial Fibrillary Acidic Protein; Head and Neck Neoplasms; HLA Antigens; Humans; Immunohistochemistry; Keratins; Neurofilament Proteins

Topics: Adenoma, Pleomorphic; Adolescent; Choristoma; Cytoplasm; Head and Neck Neoplasms; Humans; Keratins; Male; Neoplasms, Multiple Primary; Salivary Glands; Vacuoles

Malignant dermal cylindromas are very rare. We present a case of multiple cylindromas of the scalp with metastasis to a cervical lymph node. The morphology of the tumour was unusual in that it contained eccrine spiradenoma-like areas and foci of squamous differentiation with keratin formation. The immunohistochemical phenotype of the eccrine spiradenoma-like areas and the metastatic tumour was similar, but different from the areas of typical cylindroma. Although alleged "malignant" cylindromas have been reported, none have been described to have metastasized, whereas metastatic eccrine spiradenoma is well-documented. We reiterate that overlaps between dermal cylindroma and eccrine spiradenoma are more common than has been documented. In the rare event of metastases, it is the eccrine spiradenomatous component that is metastatic. We contend that there is no evidence that pure dermal cylindromas have metastasized.

Topics: Carcinoembryonic Antigen; Carcinoma, Adenoid Cystic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; S100 Proteins; Scalp Dermatoses; Skin Neoplasms; Staining and Labeling

Intermediate filament proteins are distributed in a tissue specific manner throughout human tissues. Using monoclonal or polyclonal antibodies to cytokeratins, vimentin, desmin, neurofilament proteins or the glial fibrillary acidic protein, epithelial, mesenchymal, myogenic, nervous and glial tissues, respectively, can be distinguished by immunohistochemical techniques. Since tumour cells generally retain the intermediate filament proteins typical for their cells of origin, such antibodies can also be used to discriminate between different types of neoplasma, i.e. carcinoma, lymphoma, myosarcoma, etc. Furthermore, monoclonal antibodies to individual cytokeratin proteins can be used to distinguish between several types of epithelial tissues and different types of carcinomas. The application of such antibodies in the histopathology of head and neck tumours can be of great help in the characterization of tumours that cannot be identified on the basis of routine histological techniques.

Topics: Antibodies, Monoclonal; Head and Neck Neoplasms; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratins

In head and neck squamous cell carcinoma (HNSCC), salvage neck dissection (ND) is required after primary chemoradiation in case of residual nodal disease. Upon histopathological examination, viability of tumor cells is assessed but little is known about other prognostic histopathological features. In particular, the presence of swirled keratin debris and its prognostic value is controversial. The aim of this study is to examine histopathological parameters in ND specimens and correlate them with patient outcome to determine the relevant parameters for histopathological reporting.. Salvage ND specimen from a cohort of n = 75 HNSCC (oropharynx, larynx, hypopharynx) patients with prior (chemo) radiation were evaluated on H&E stains for the following parameters: viable tumor cells, necrosis, swirled keratin debris, foamy histiocytes, bleeding residues, fibrosis, elastosis, pyknotic cells, calcification, cholesterol crystals, multinucleated giant cells, perineural, and vascular invasion. Histological features were correlated with survival outcomes.. Only the presence / amount (area) of viable tumor cells correlated with a worse clinical outcome (local and regional recurrence-free survival, (LRRFS), distant metastasis-free survival, disease-specific survival, and overall survival, p < 0.05) in both the univariable and multivariable analyses.. We could confirm the presence of viable tumor cells as a relevant negative prognostic factor after (chemo) radiation. The amount (area) of viable tumor cells further substratified patients with worse LRRFS. None of the other parameters correlated with a distinctive worse outcome. Importantly, the presence of (swirled) keratin debris alone should not be considered viable tumor cells (ypN0).

Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Keratins; Lymphatic Metastasis; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; Retrospective Studies; Squamous Cell Carcinoma of Head and Neck

Oral squamous cell carcinoma (OSCC) is one of the most common malignancies affecting the head-and-neck region, regional lymph nodes being an important prognostication factor dictating the survival rate. Despite an array of modalities used, clinically, radiographically and routine histopathologically, the detection of micro-metastasis (2-3 mm tumour cell deposits) in the lymph nodes often escapes identification. The presence of few of these tumour epithelial cells in the lymph nodes drastically increases mortality and alters treatment plan. Hence, the identification of these cells is of major prognostic significance for a patient. Thus, the present study was aimed to evaluate and detect the efficacy of the immunohistochemical (IHC) marker [cytokeratin (CK) AE1/AE3] over routine Hematoxylin & eosin (H & E) staining in detecting micro-metastasis in the lymph nodes of OSCC cases.. The IHC marker CK cocktail (AE1/AE3) did not demonstrate any positive reactivity for the target antigen in all the 100 H & E stained lymph node sections evaluated in the present study.. This study was undertaken to check the efficacy of IHC (CK cocktail AE1/AE3) in the detection of micro-metastasis in lymph nodes that are found to be negative in routine H&E stained sections. The findings of this study suggest that the IHC marker AE1/AE3 did not prove to be useful to detect micro-metastasis in this study population.

Topics: Carcinoma, Squamous Cell; Eosine Yellowish-(YS); Head and Neck Neoplasms; Hematoxylin; Humans; Immunohistochemistry; Keratins; Lymph Nodes; Lymphatic Metastasis; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

Topics: Adamantinoma; Adolescent; Adult; Ameloblastoma; Biomarkers, Tumor; Child; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; RNA-Binding Protein EWS; Sarcoma, Ewing; Young Adult

The archetypal solitary fibrous tumor (SFT) features fibroblastic cells with varying cellularity without any particular architectural arrangement in a collagenous matrix, with staghorn vessels, CD34 and STAT6 expression, and NAB2::STAT6. To date, this fusion is thought to be specific for SFT. With more routine use of fusion gene panels, the histologic diversity of NAB2::STAT6-positive tumors is increasingly appreciated. Here we present four head and neck tumors harboring NAB2::STAT6 but exhibiting remarkably unusual morphologic features for SFT. All cases were pulled from the authors' consultation files. Immunohistochemistry was performed, along with targeted RNA sequencing in all cases, plus DNA next-generation sequencing on two. The cases arose in the nasal cavity (n = 2), retromolar trigone (n = 1) and parapharynx (n = 1), in patients ranging from 39 to 54 (mean, 44). Both nasal cases were biphasic, with a variably cellular collagenized stroma that resembled SFT but also interspersed malignant epithelial and neuroepithelial nests. One of the nasal cases also exhibited overt rhabdomyoblastic differentiation within both components. The two non-nasal cases were comprised of plump, epithelioid cells that were diffusely positive for pan-cytokeratin. One of these cases had prominent cystic change lined by overtly squamous epithelium. STAT6 immunostaining was positive in all cases, although the epithelial/neuroepithelial nests in the sinonasal cases were negative. All cases were confirmed to harbor NAB2::STAT6 by RNA sequencing. The two sinonasal cases were also found to harbor oncogenic mutations. The presented cases highlight a much broader histologic diversity than previously known for neoplasms with NAB2::STAT6. The biphasic nasal cases closely resemble teratocarcinosarcoma, while the epithelioid, cytokeratin-positive cases could be conceptualized as "adamantinoma-like," to borrow terminology already in use for Ewing sarcomas with complex epithelial differentiation. To identify similar cases, pathologists should have a low threshold for using STAT6 immunohistochemistry on any difficult-to-characterize head and neck tumor.

Topics: Adamantinoma; Ameloblastoma; Biomarkers, Tumor; Head and Neck Neoplasms; Humans; Keratins; Repressor Proteins; Solitary Fibrous Tumors; STAT6 Transcription Factor

We investigated whether in human head and neck squamous cell carcinoma (HNSCC) high levels of expression of stress keratin 17 (K17) are associated with poor survival and resistance to immunotherapy.. We investigated the role of K17 in regulating both the tumor microenvironment and immune responsiveness of HNSCC using a syngeneic mouse HNSCC model, MOC2. MOC2 gives rise to immunologically cold tumors that are resistant to immune-checkpoint blockade (ICB). We engineered multiple, independent K17 knockout (KO) MOC2 cell lines and monitored their growth and response to ICB. We also measured K17 expression in human HNSCC of patients undergoing ICB.. MOC2 tumors were found to express K17 at high levels. When knocked out for K17 (K17KO MOC2), these cells formed tumors that grew slowly or spontaneously regressed and had a high CD8+ T-cell infiltrate in immunocompetent syngeneic C57BL/6 mice compared with parental MOC2 tumors. This phenotype was reversed when we depleted mice for T cells. Whereas parental MOC2 tumors were resistant to ICB treatment, K17KO MOC2 tumors that did not spontaneously regress were eliminated upon ICB treatment. In a cohort of patients with HNSCC receiving pembrolizumab, high K17 expression correlated with poor response. Single-cell RNA-sequencing analysis revealed broad differences in the immune landscape of K17KO MOC2 tumors compared with parental MOC2 tumors, including differences in multiple lymphoid and myeloid cell types.. We demonstrate that K17 expression in HNSCC contributes to immune evasion and resistance to ICB treatment by broadly altering immune landscapes of tumors.

Topics: Animals; Head and Neck Neoplasms; Humans; Immune Checkpoint Inhibitors; Immune Evasion; Keratin-17; Keratins; Mice; Mice, Inbred C57BL; Squamous Cell Carcinoma of Head and Neck; Tumor Microenvironment

Perineural invasion (PNI), a common occurrence in oral squamous cell carcinomas, is associated with poor survival. Consequently, these tumors are treated aggressively. However, diagnostic criteria of PNI vary and its role as an independent predictor of prognosis has not been established. To address these knowledge gaps, we investigated spatial and transcriptomic profiles of PNI-positive and PNI-negative nerves.. Tissue sections from 142 patients were stained with S100 and cytokeratin antibodies. Nerves were identified in two distinct areas: tumor bulk and margin. Nerve diameter and nerve-to-tumor distance were assessed; survival analyses were performed. Spatial transcriptomic analysis of nerves at varying distances from tumor was performed with NanoString GeoMx Digital Spatial Profiler Transcriptomic Atlas.. PNI is an independent predictor of poor prognosis among patients with metastasis-free lymph nodes. Patients with close nerve-tumor distance have poor outcomes even if diagnosed as PNI negative using current criteria. Patients with large nerve(s) in the tumor bulk survive poorly, suggesting that even PNI-negative nerves facilitate tumor progression. Diagnostic criteria were supported by spatial transcriptomic analyses of >18,000 genes; nerves in proximity to cancer exhibit stress and growth response changes that diminish with increasing nerve-tumor distance. These findings were validated in vitro and in human tissue.. This is the first study in human cancer with high-throughput gene expression analysis in nerves with striking correlations between transcriptomic profile and clinical outcomes. Our work illuminates nerve-cancer interactions suggesting that cancer-induced injury modulates neuritogenesis, and supports reclassification of PNI based on nerve-tumor distance rather than current subjective criteria.

Topics: Head and Neck Neoplasms; Humans; Keratins; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Staging; Peripheral Nerves; Prognosis; Retrospective Studies; Transcriptome

Topics: Cadherins; Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Head and Neck Neoplasms; Humans; Keratins; Mitochondria; Oligomycins; Vimentin

Extracellular vesicles (EVs) have recently come into the spotlight as potential cancer biomarkers. Isolation of pure EVs is complex, so wider use requires reliable and time‑efficient isolation methods. In the present study, galectin‑based magnetic glycan recognition particles, EXÖBead

Topics: B7-H1 Antigen; Biomarkers, Tumor; Epithelial Cell Adhesion Molecule; Extracellular Vesicles; Galectins; Head and Neck Neoplasms; Humans; Keratins; Leukocytes, Mononuclear; Lipoproteins; Polyethylene Glycols; Polysaccharides; Squamous Cell Carcinoma of Head and Neck

Fanconi anemia (FA) patients frequently develop oral squamous cell carcinoma (OSCC). This cancer in FA patients is diagnosed within the first 3-4 decades of life, very often preceded by lesions that suffer a malignant transformation. In addition, they respond poorly to current treatments due to toxicity or multiple recurrences. Translational research on new chemopreventive agents and therapeutic strategies has been unsuccessful partly due to scarcity of disease models or failure to fully reproduce the disease. Here we report that Fanca gene knockout mice (Fanca

Topics: Animals; Carcinoma, Squamous Cell; Disease Models, Animal; Fanconi Anemia; Head and Neck Neoplasms; Keratins; Mice; Mice, Knockout; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

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

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

We describe a cytokeratin positive interstitial reticulum cell tumor (CPIRCT) as the cause of a large and defacing scalp tumor. Clinically these tumors manifest as progressive, painless swelling. Treatment usually consists of surgery with or without irradiation; chemotherapy is applied in metastatic disease.. A patient was referred after attempted removal of a large bump on the head. The tumor was initially noted about 12 months earlier. Assuming a benign lipoma, resection without prior imaging was attempted. During surgery, the underlying bone was found to be profoundly destroyed. Cranial magnetic resonance imaging revealed a large mass with an extracranial and intracranial component. Subsequent extensive resection finally led to the diagnosis of CPIRCT.. Most CPIRCTs manifest as progressive palpable or visible masses. Radical excision is usually the mainstay of treatment, although there is no generally accepted treatment strategy. A needle biopsy might not be diagnostic and can complicate future curative surgery. Especially in fast-growing lesions, imaging studies should be considered before surgery. Their potential recurrence and metastatic spread render CPIRCTs an interdisciplinary challenge and highlight the need for long-term follow-up.

Topics: Adult; Brain Neoplasms; Diagnostic Errors; Head and Neck Neoplasms; Humans; Keratins; Lipoma; Magnetic Resonance Imaging; Male; Scalp; Skull Neoplasms

Adamantinoma-like Ewing Sarcoma (ALES) is a rare subtype of Ewing sarcoma family of tumors (EFTs) which are defined by their EWSR1 gene rearrangements. We present a case of a 15-year old female with a swelling in her anterior neck of 4 months duration which had recently begun to rapidly grow in size. Fine needle aspiration showed a small blue round cell tumor with immunoreactivity for cytokeratin, CD99 and FLI1. Material for molecular testing was available on the resection specimen. Demonstration of t(11;22) (EWS-FLI1) was helpful in establishing the diagnosis.

Topics: 12E7 Antigen; Adamantinoma; Adolescent; Biomarkers, Tumor; Biopsy, Fine-Needle; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; In Situ Hybridization, Fluorescence; Keratins; Oncogene Proteins, Fusion; Proto-Oncogene Protein c-fli-1; RNA-Binding Protein EWS; Sarcoma, Ewing; Thyroid Gland; Thyroidectomy

Tumor budding (TB) is a promising prognostic marker in many cancers including oral squamous cell carcinoma. The evaluation of TB in preoperative diagnostic biopsies has been proven be possible; therefore, the association of TB with other morphological features can represent an important aid in the previous treatment decision. This study aims to evaluate TB in oral squamous cell carcinoma (OSCC) biopsies, assessing its association with other morphological characteristics of the sample. A total of 56 cases of OSCC were investigated. In hematoxylin and eosin-stained slides, morphological features including histopathological grading and mode of invasion were evaluated in the deep invasive front. Moreover, immunohistochemistry was performed with anti-multi-cytokeratin antibody helping in the identification of TB, which was graded as low-intensity or no TB and high-intensity TB. Descriptive and bivariate analyses were performed, and the level of significance was set at 5%. The tongue was the most-affected site with 29 (51.7 %) tumors. The predominant mode of invasion (27-48.2 %) was by groups of neoplastic cells without clear boundaries. Of the cases investigated, 37 (66.1 %) were high-intensity TB, which was associated with the mode of invasion of the tumors (p < 0.05). All cases with the worst mode of invasion showed high-intensity TB. Preliminary results showed the potential of morphological features, such as TB and mode of invasion, evaluated in diagnostic specimens of OSCC, aiding in the treatment decision to select patients who could benefit from more-aggressive treatments.

Topics: Adult; Biopsy; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Squamous Cell Carcinoma of Head and Neck

Desmoplastic melanoma (DM) is an uncommon variant of malignant melanoma (MM), histologically characterized by a mainly dermal proliferation of spindled cells within a desmoplastic stroma. Normally, involvement of deeper tissues by DM is the result of direct extension down from the overlying dermis. MM is widely known to harbor a striking potential for morphological and phenotypic variability; among MM morphological variants, pseudoglandular MM is characterized by extensive discohesion within cords and nests of malignant cells and ensuing formation of so-called pseudolumina, thus mimicking adenocarcinoma. We present an exceptional case of DM characterized by intrafascial origin, partly pseudoglandular differentiation, and aberrant experession of cytokeratins in the pseudoglandular component; genetic data from next-generation sequencing supported the final diagnosis of DM, as well as the ontogenetic identity of the pseudoglandular component. Prior to this report, pseudoglandular features had never been described in DM. Additionally, our case is unusual because of the deep origin of the tumor, arising below the subcutaneous fat of the scalp, as well as the aberrant experession of cytokeratins in the pseudoglandular component, thus posing a challenging differential diagnosis with several soft tissue tumors.

Topics: Aged; Biomarkers, Tumor; Cell Differentiation; Diagnosis, Differential; Head and Neck Neoplasms; Humans; Keratins; Male; Melanoma; Predictive Value of Tests; Scalp; Skin Neoplasms

Bone morphogenetic protein 6 (BMP6) has unique properties regarding structure and function in supporting bone formation during development and adult life. Despite its known role in various malignant tumors, the prognostic significance of BMP6 expression in oral squamous cell carcinoma (OSCC) remains unknown. The aim of the study was to investigate immunohistochemical expression of BMP6 in OSCC in correlation with clinical and pathological parameters, disease recurrence and survival. In addition, we investigated other parameters in order to identify prognosticators of neck metastases and final outcome. The study included 120 patients with clinically T1-3N0 OSCC who were primarily surgically treated between 2003 and 2008. There were 99 (82.5%) male and 21 (17.5%) female patients. The five-year disease-specific survival for the whole cohort was 79.7%. Tumors smaller than 2 cm in diameter showed higher incidence of strong BMP6 expression. No statistical correlation was observed between other clinico-pathological factors and BMP6 expression. Expression of BMP6 was not associated with disease recurrence and survival. BMP6 may not serve as prognosticator of final outcome or recurrence in clinically node-negative OSCC subjects. In multivariate analysis predictors of poorer survival were positive surgical margin, moderate tumor cell differentiation and pathological involvement of levels IV and/or V.

Topics: Adult; Aged; Aged, 80 and over; Bone Morphogenetic Protein 6; Carcinoma, Squamous Cell; Cohort Studies; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mouth Neoplasms; Multivariate Analysis; Neoplasm Metastasis; Neoplasm Recurrence, Local; Prognosis

Occult neck metastasis is an important prognostic factor in patients with tongue squamous cell carcinoma (TSCC) who are deemed clinically negative for neck metastasis. The purpose of this study was to identify predictive factors for occult neck metastasis arising from TSCC and to determine patient prognosis. Ninety-seven patients with cT2N0 TSCC who underwent surgical resection of their primary lesion as initial therapy were enrolled in this retrospective study. Cutoff values for depth of invasion (≥3.3 mm) and the tumor budding score (≥4) were determined using receiver operator characteristic analyses. Univariate and multivariate analyses revealed that a tumor budding score ≥4 is a significant independent predictive factor for the occurrence of occult neck metastasis, which in turn is a significant independent prognostic factor. When evaluating tumor budding, we demonstrated greater interobserver and intraobserver agreement when using immunohistochemical staining for cytokeratin AE1/AE3 than with hematoxylin and eosin staining (HE). We conclude that the evaluation of tumor budding is effective for identifying populations at high risk of occult neck metastasis, which will enable the planning of appropriate therapeutic strategies for patients with cT2N0 TSCC. Furthermore, cytokeratin staining is recommended over HE staining for simpler and more accurate evaluation of tumor budding.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Cell Movement; Female; Glossectomy; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Observer Variation; Predictive Value of Tests; Reproducibility of Results; Retrospective Studies; Squamous Cell Carcinoma of Head and Neck; Staining and Labeling; Tongue Neoplasms; Treatment Outcome

: TRIM29 (tripartite motif-containing protein 29) is a TRIM family protein that has been implicated in breast, colorectal, and pancreatic cancers. However, its role in stratified squamous epithelial cells and tumors has not been elucidated. Here, we investigate the expression of TRIM29 in cutaneous head and neck squamous cell carcinomas (SCC) and its functions in the tumorigenesis of such cancers. TRIM29 expression was lower in malignant SCC lesions than in adjacent normal epithelial tissue or benign tumors. Lower expression of TRIM29 was associated with higher SCC invasiveness. Primary tumors of cutaneous SCC showed aberrant hypermethylation of

Topics: Animals; Biomarkers, Tumor; Carcinogenesis; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Movement; Cell Proliferation; DNA Methylation; DNA-Binding Proteins; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Keratinocytes; Keratins; Mice; Mice, Nude; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasm Transplantation; Prognosis; Proteins; RNA, Small Interfering; Transcription Factors

The expression of the hair follicle stem cell marker CD34 was analyzed in five different head and neck squamous cell carcinoma (HNSCC) cell lines with different antibodies. All HNSCC cell lines expressed CD34 on their cell surface. After cell cycle synchronization via serum starvation, we observed cyclic CD34 expression in HNSCC cells dependent on cell cycle progression via immunofluorescent staining and flow cytometric analysis. Investigation of the CD34(+) and CD34(-) HNSCC populations revealed most of the cells in S-phase and G2/M-Phase in CD34(+) cells in contrast to CD34(-) cells. Knockdown of CD34 in HNSCC cells led to diminished clonal expansion in a colony forming assay after subjecting the cells to ionizing radiation. Furthermore, knockdown of CD34 after cell cycle synchronization induced high CK1, CK4, and CK5 gene expression and downregulation of CK10 gene expression as shown by Taqman

Topics: Antigens, CD34; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Gene Expression; Head and Neck Neoplasms; Humans; Keratins; Radiation Tolerance

To investigate the immunohistochemical expression (IHCE) of selected keratins in primary cutaneous and mucosal melanoma (pM), and metastatic melanoma (metsM) of the head and neck and to compare their expression to a group of undifferentiated/poorly differentiated tumors of the same anatomic region.. IHCE of K6, K7, K8, K14, K16, K18, and K19 were studied in 29 melanomas and 70 cases of non-melanoma tumors of the same anatomic region (neuroendocrine carcinoma, neuroblastoma, olfactory neuroblastoma, sinonasal undifferentiated carcinoma, undifferentiated nasopharyngeal carcinoma, anaplastic large cell lymphoma, poorly differentiated squamous cell carcinoma (PDSCC), and Ewing sarcoma). MNF-116 pancytokeratin was investigated in melanoma.. All studied keratins, except K6, were expressed in melanoma. IHCE of MNF-116, K8, and K18 was higher in metsM compared with pM. K14 and K16 expression was highest in PDSCC.. metsM expresses keratins more than pM, specifically K8, K18, and MNF-116. Keratin positivity in an undifferentiated or poorly differentiated neoplasm does not necessarily exclude the diagnosis of melanoma.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Male; Melanoma; Middle Aged

Patients with head and neck squamous cell carcinoma (HNSCC) present different responses to chemotherapy and radiotherapy. One explanation may be the differences in the individual rates of stem cell-like cells.. We included patients with HNSCC and tumor progression or relapse. Tumor samples were obtained before and after primary chemotherapy, and immunohistochemical analyses were performed for CD44, HLA class I (HLA-I), pancytokeratin, and phosphorylated epidermal growth factor receptor (p-EGFR). Differences in expression between the first and second specimens were assessed.. Expression between the first and second specimens varied as follows: CD44 increased by 14.67% (95% confidence interval, CI: 6.94 to 22.40; p < 0.01); HLA-I decreased by 16.72% (95% CI: -23.87 to -9.47; p < 0.01); pancytokeratin decreased by 24.91% (95% CI: -32.8 to -17.7; p < 0.01), and p-EFGR expression decreased by 12.30% (95% CI: -20.61 to -3.98; p < 0.005).. Among patients with HNSCC, there is an enrichment of cells with stem-like markers in relapsed tumors when compared with the primary tumor. This finding should be considered when developing treatment strategies.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease Progression; ErbB Receptors; Female; Head and Neck Neoplasms; Histocompatibility Antigens Class I; Humans; Hyaluronan Receptors; Keratins; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplastic Stem Cells

Squamous cell carcinoma (SSC) of the head and neck is the sixth most common cancer and is rarely diagnosed in early stages. The transcription factor Krϋppel-like factor 4 (Klf4) suppresses cell proliferation and promotes differentiation. Inducible mice carrying an oral-specific ablation of Klf4 (K14-CreER(tam) /Klf4(flox/flox) ) develop mild dysplastic lesions and abnormal differentiation in the tongue. Aiming to analyze whether Klf4 cooperate in oral chemical carcinogenesis,we applied 4-nitroquinoline 1-oxide (4NQO), a tobacco surrogate, to this conditional Klf4 knockout mice.. K14-CreER(tam) /Klf4(flox/flox) and control mice were treated with 4NQO for 16 weeks and monitored until week 30. Histopathological samples were used for diagnostic purposes and immunofluorescence detection of epithelial differentiation markers.. 4NQO-treated K14-CreER(tam) /Klf4(flox/flox) mice (Klf4KO 4NQO) showed a significant weight loss and developed more severe dysplastic lesions than control mice with 4NQO (P < 0.005). The Klf4KO 4NQO showed a tendency to higher incidence of oral SCC and a marked keratinization pattern in dysplasias, in situ carcinomas and SCC. Also, tongues derived from Klf4KO 4NQO mice exhibited reduced terminal differentiation as judged by cytokeratin 1 staining when compared with 4NQO-treated controls.. Klf4 ablation results in more severe dysplastic lesions in oral mucosa, with a tendency to higher incidence of SCC, after chemical carcinogenesis. We show here, in a context similar to the human carcinogenesis, that absence of Klf4 accelerates carcinogenesis and correlates with the absence of cytokeratin 1 expression. These results suggest a potential role for KLF4 as a tumor suppressor gene for the tongue epithelium.

Topics: 4-Nitroquinoline-1-oxide; Animals; Carcinogenesis; Carcinogens; Carcinoma, Squamous Cell; Cell Differentiation; Disease Models, Animal; Gene Expression; Head and Neck Neoplasms; Keratins; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Mice; Mice, Inbred C57BL; Mice, Knockout; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Squamous Cell Carcinoma of Head and Neck; Tongue Neoplasms

The invasion of cancer cells into the surrounding normal tissue is one of the defining features of cancer. While the phenomena of tumour budding, epithelial-mesenchymal transition and the presence of myofibroblasts have independently been shown to be related to a poor prognosis of oral carcinomas, their relationship has not been examined in detail.. Paraffin-embedded tissues from 28 patients with oral squamous cell carcinomas were stained with antibodies to cytokeratin, α-SMA, vimentin, E-cadherin, N-cadherin and Twist and evaluated for their expression in relation to invasive cancer cells and the surrounding tumour stroma.. A direct, histological relationship between invading, budding tumour cells and myofibroblasts was occasionally seen but was not a general feature. Most of the budding tumour cells at the invasive front had a decreased expression of E-cadherin, but we did not find that this was associated with a consistent or clear increase in either N-cadherin or vimentin. We therefore suggest that the budding of tumour cells is not dependent upon either myofibroblasts or a complete epithelial-mesenchymal transition and that these phenomena most likely represent separate processes in tumour progression.

Topics: Actins; Animals; Cadherins; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; Goats; Head and Neck Neoplasms; Humans; Keratins; Mice; Mouth Neoplasms; Muscle, Smooth; Myofibroblasts; Paraffin Embedding; Prognosis; Rabbits; Single-Cell Analysis; Squamous Cell Carcinoma of Head and Neck; Twist-Related Protein 1; Vimentin

Ewing sarcoma family tumors (EFTs) of the head and neck are rare and may be difficult to diagnose, as they display significant histologic overlap with other more common undifferentiated small blue round cell malignancies. Occasionally, EFTs may exhibit overt epithelial differentiation in the form of diffuse cytokeratin immunoexpression or squamous pearls, resembling the so-called adamantinoma-like EFTs and being challenging to distinguish from bona fide carcinomas. Furthermore, the presence of EWSR1 gene rearrangement correlated with strong keratin expression may suggest a myoepithelial carcinoma. Herein, we analyze a series of 7 adamantinoma-like EFTs of the head and neck, most of them being initially misdiagnosed as carcinomas because of their anatomic location and strong cytokeratin immunoexpression, and subsequently reclassified as EFT by molecular techniques. The tumors arose in the sinonasal tract (n=2), parotid gland (n=2), thyroid gland (n=2), and orbit (n=1), in patients ranging in age from 7 to 56 years (mean, 31 y). Microscopically, they departed from the typical EFT morphology by growing as nests with peripheral nuclear palisading and prominent interlobular fibrosis, imparting a distinctly basaloid appearance. Moreover, 2 cases exhibited overt keratinization in the form of squamous pearls, and 1 sinonasal tumor demonstrated areas of intraepithelial growth. All cases were positive for CD99, pancytokeratin, and p40. A subset of cases showed synaptophysin, S100 protein, and/or p16 reactivity, further confounding the diagnosis. Fluorescence in situ hybridization assays showed EWSR1 and FLI1 rearrangements in all cases. Our results reinforce that a subset of head and neck EFTs may show strong cytokeratin expression or focal keratinization, and are therefore histologically indistinguishable from more common true epithelial neoplasms. Thus, CD99 should be included in the immunopanel of a round cell malignancy regardless of strong cytokeratin expression or anatomic location, and a strong and diffuse CD99 positivity should prompt molecular testing for the presence of EWSR1 gene rearrangements.

Topics: 12E7 Antigen; Adamantinoma; Adolescent; Adult; Antigens, CD; Biomarkers, Tumor; Biopsy; Bone Neoplasms; Calmodulin-Binding Proteins; Cell Adhesion Molecules; Cell Differentiation; Child; Diagnosis, Differential; Female; Gene Rearrangement; Head and Neck Neoplasms; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratins; Male; Middle Aged; Myoepithelioma; Neuroectodermal Tumors, Primitive, Peripheral; Predictive Value of Tests; Proto-Oncogene Protein c-fli-1; RNA-Binding Protein EWS; RNA-Binding Proteins; Sarcoma, Ewing; Tissue Array Analysis; Young Adult

Sarcomatoid carcinoma (SC) is a variant of head and neck squamous cell carcinoma characterized by a prominent and sometimes exclusive spindle cell component. Distinction from a sarcoma or reactive stroma can be problematic, particularly in cases in which the conventional component is not obvious. The value of immunohistochemistry is limited because of the loss of cytokeratin expression in a sizable percentage of cases. Staining for p63 can enhance detection of epithelial differentiation, but its usefulness is offset by expression in various soft tissue proliferations. Staining for p40--a squamous-specific isoform of p63--could potentially improve diagnostic accuracy. Immunohistochemistry for pancytokeratin, p63, and p40 was performed on 37 head and neck SCs, 201 soft tissue neoplasms, and 40 reactive stromal proliferations. The SCs were also stained for p16 in the event that some of the tumors were human papillomavirus (HPV) related. HPV in situ hybridization was performed on p16-positive cases. Twenty-three of 37 (62%) SCs were positive for pancytokeratin, 23 of 37 (62%) were positive for p63, and 20 of 37 (54%) were positive for p40. Compared with p63, p40 staining was less likely to be observed in soft tissue tumors (5% vs. 30%) and reactive stromal proliferations (0% vs. 30%). HPV16 was detected in 3 of 10 (30%) SCs of the oropharynx but in none of the nonoropharyngeal SCs. p40 staining does not improve the sensitivity for diagnosing SC, but it does diminish the risk of misdiagnosing a sarcoma or reactive stroma as SC. The presence of a sarcomatoid variant of HPV-related oropharyngeal cancer points to HPV testing as a useful diagnostic tool for atypical spindle cell proliferations of the oropharynx.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p16; Diagnosis, Differential; DNA, Viral; Head and Neck Neoplasms; Human papillomavirus 16; Human Papillomavirus DNA Tests; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Oropharyngeal Neoplasms; Papillomavirus Infections; Predictive Value of Tests; Protein Isoforms; Stromal Cells; Transcription Factors; Tumor Suppressor Proteins

The extracapsular spread (ECS) of lymph node metastasis (LNM) reflects tumor aggressiveness and is associated with poor survival and risk of distant metastasis. In this study, we aimed to explore the prognostic significance of epithelial-mesenchymal transition (EMT) of ECS tumors in LNM of head and neck cancers.. We collected LNM samples from head and neck cancer patients (follow-up >2 years) and made 20 ECS(-): ECS(+) pairs (1:2) of LNM (N = 60), matched by the primary sites and by T and N classifications. Immunostaining of cytokeratin, E-cadherin, vimentin, and CD31 were performed and quantified to determine the epithelial-mesenchymal transition percent (EMT%), defined as vimentin(+)/cytokeratin(+) area of ECS. Univariate and multivariable analyses of clinic-pathologic factors, including EMT% of ECS, were conducted to identify the significant prognosticators. In addition, the anatomical relationship between CD31 vessels and ECS tumors was analyzed.. Rather than the presence of ECS in LNM, higher EMT% (>50 %) of ECS strongly correlated with the worse overall and disease-free survival and had more frequent recurrence and distant dissemination in their clinical courses. ECS tumors intermingled closely with Ki-67(-) CD31(+) non-proliferating perinodal blood vessels. Particularly, vimentin(+) ECS areas exhibited a higher density of CD31(+) perinodal vessels than did vimentin(-) ECS.. High EMT scores of ECS tumors in LNM predict an unfavorable prognosis and systemic dissemination more accurately than the simple presence of ECS in LNM in head and neck cancer patients.

Topics: Cadherins; Disease-Free Survival; Epithelial-Mesenchymal Transition; Female; Head and Neck Neoplasms; Humans; Keratins; Ki-67 Antigen; Lymph Nodes; Lymphatic Metastasis; Male; Neoplasm Staging; Platelet Endothelial Cell Adhesion Molecule-1; Survival Rate; Vimentin

The objective of this study was to evaluate the potential detection of circulating tumor cells (CTCs) using the CellSearch (CS) Assay™ in patients with locally advanced head and neck squamous cell carcinoma (HNSCC) and then to identify the clinical factors predictive of the presence of CTCs. The presence and number of CTCs were determined using the CS system before treatment, and in 10 healthy individuals (control group). The CS system was able to successfully identify the presence of CTCs in 8 of 49 patients (16 %) before therapy. No CTC was found in the control group. CTCs were detected before therapy in 1 of 19 patients (5 %) with N0 tumor and in 7 of 30 patients (23 %) with N1-2c tumor (p = 0.12; Fisher's exact test). CTCs were identified in a relatively low proportion of patients with locally advanced HNSCC.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Cell Adhesion Molecules; Cell Count; Epithelial Cell Adhesion Molecule; Female; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Neoplastic Cells, Circulating; Oropharyngeal Neoplasms; Prognosis; Squamous Cell Carcinoma of Head and Neck

The hedgehog signaling pathway (HH) is involved in tumorigenesis in a variety of human malignancies. In head and neck squamous cell carcinomas (HNSCC), Hh overexpression was associated with poor prognosis. Therefore, we analyzed the effect of Hh signaling blockade with cyclopamine on colony formation of cells from HNSCC samples.. HNSCC biopsies were cultured alone for reference or with serial dilutions of cyclopamine (5-5,000 nM), docetaxel (137.5-550 nM), or cisplatin (1,667-6,667 nM) and their binary combinations. Cytokeratin-positive colonies were counted after fluorescent staining.. Cyclopamine concentration-dependently inhibited HNSCC ex vivo [(IC50) at about 500 nM]. In binary combinations, cyclopamine additively enhanced the suppressive effects of cisplatin and docetaxel on HNSCC colony formation.. Our findings define SMO--a Hh component- as a potential target in HNSCC and suggest the utility of Hh targeting in future multimodal treatment regimens for HNSCC.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Carcinoma, Squamous Cell; Cisplatin; Docetaxel; Female; Head and Neck Neoplasms; Hedgehog Proteins; Humans; KB Cells; Keratins; Male; Molecular Targeted Therapy; Receptors, G-Protein-Coupled; Signal Transduction; Smoothened Receptor; Taxoids; Tumor Cells, Cultured; Tumor Stem Cell Assay; Veratrum Alkaloids

Keratins play a major role in several cellular functions. Each tissue type expresses a specific set of keratins. The immense potential of keratins as diagnostic and prognostic markers for different cancers is emerging. Oral cancer is the fifteenth most common cancer worldwide. However, comprehensive information on the profile of keratins in the oral cavity is not available. Several independent reports have identified keratins using antibody based techniques which have pitfalls due to the cross reactivity of the antibodies to this set of very homologous proteins. A few recent proteomic studies have reported the identification of keratins in head and neck cancer. Majority of the studies have used tissues from the head and neck region without specifying subsites. This study reports the analysis of enriched preparations of keratins from cancer of the gingivo buccal complex (GBC) using MS, 2DE, WB, silver staining of 2DE gels and IHC. Our study reveals the absence of K4 and K13 and presence of K14, K16, and K17, in cancers of the GBC and combination of these expression patterns in the cut margins. This report also shows that K13 is glycosylated. This well characterized profile of keratins may have potential to be used in clinics.. In recent years the immense potential of keratins as diagnostic and prognostic markers for different cancers is emerging. However, comprehensive information on the profile of keratins in the oral cavity is not available. Several independent reports have identified keratins using only antibody based techniques which have pitfalls due to the cross reactivity of the antibodies to this set of very homologous proteins. This study reports the analysis of enriched preparations of keratins from a subsite of the oral cavity, the gingivo buccal complex (GBC) using mass spectrometry, 2DE, western blotting, silver staining of 2DE gels and IHC. The proteomic analysis shows the absence of K4 and K13 and presence of K14, K16, and K17 in cancers of the GBC and combination of these expression patterns in the cut margins. This well characterized profile of keratins from the gingivo buccal complex provides defined markers which may have potential to be used in the clinics.

Topics: Adult; Aged; Biomarkers; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Gingiva; Glycosylation; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Neoplasm Metastasis; Proteomics

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

The objective of this study is to evaluate the incidence of occult nodal micrometastases in N0 laryngeal squamous cell carcinoma using cytokeratin immunohistochemical analysis (CKIHA) and its influence on the overall occult metastatic rate. This is a prospective cohort study. A total number of 30 patients with N0 stage laryngeal cancer underwent 46 selective neck dissections for elective treatment of the neck. Nodes found to be negative using routine histopathological examination were evaluated for the presence of micrometastasis using CKIHA. The occult micrometastasis rate for all cases was 26.7 % which significantly increased the overall occult metastasis rate to 50 % (P = 0.014). The micrometastasis rate was 30.8, 25 and 20 % for glottic, supraglottic and transglottic tumors, respectively, which increased the overall occult metastasis rate to 53.8, 50 and 40 % but without statistical impact. The micrometastasis rate was 35.7 and 23.1 % in T3 and T4 tumors, respectively, and this increased the overall occult metastasis rate to 50 and 61.5 % with statistical influence in T3 tumors (P = 0.046). Micrometastasis upstaged the neck in 7 (23.3 %) patients with statistical impact on the PN stage (P = 0.007). The overall occult nodal metastasis rate in N0 laryngeal cancer is underestimated. Nodal micrometastasis may be missed during routine histopathological examination and can be detected using CKIHA.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cohort Studies; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Neck Dissection; Neoplasm Micrometastasis; Neoplasm Staging; Prospective Studies; Sensitivity and Specificity; Squamous Cell Carcinoma of Head and Neck

The migratory ability of tumor cells requires cytoskeletal rearrangement processes. Epidermal growth factor receptor (EGFR)-signaling tightly correlates with tumor progression in head and neck squamous cell carcinomas (HNSCCs), and has previously been implicated in the regulation of cytokeratin (CK) expression. In this study, HNSCC cell lines were treated with EGF, and CK expression levels were monitored by Western blot analysis. Changes in cellular morphology were documented by fluorescence- and atomic force microscopy. Some of the cell lines demonstrated an EGF-dependent modulation of CK expression levels. Interestingly, regression of some CK subtypes or initial up-regulation followed by downregulation at higher EGF-levels could also be observed in the tested cell lines. Overall, the influence of EGF on CK expression levels appeared variable and cell-type-dependent. Real-time cellular analysis of EGF-treated and -untreated HNSCC cell lines demonstrated a rise over time in cellular impedance. In three of the EGF-treated HNSCC cell lines, this rise was markedly higher than in untreated controls, whereas in one of the cell lines the gain of cellular impedance was paradoxically reduced after EGF treatment, which was found to correlate with changes in cellular morphology rather than with relevant changes in cellular viability or proliferation. After treating HNSCC cells with EGF, CK filaments frequently appeared diffusely distributed throughout the cytoplasm, and in some cases were found in a perinuclear localization, the latter being reminiscent to observations by other groups. In summary, the data points to a possible role of EGFR in modulating HNSCC cell morphology.

Topics: Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Shape; Cell Survival; Epidermal Growth Factor; Head and Neck Neoplasms; Humans; Keratinocytes; Keratins; Microscopy, Atomic Force; Phenotype; Plakophilins; Squamous Cell Carcinoma of Head and Neck

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Bone Marrow; Carcinoma, Squamous Cell; Cell Adhesion Molecules; Epithelial Cell Adhesion Molecule; Female; Head and Neck Neoplasms; Humans; Keratins; Lymph; Lymph Nodes; Lymphatic Metastasis; Male; Microscopy, Fluorescence; Middle Aged; Polymerase Chain Reaction; Reproducibility of Results

To determine the feasibility of viable storage of head and neck squamous cell carcinoma (HNSCC) for regrowth of cells in culture.. Laboratory-based translational study.. Methods for intermediate-term frozen storage of viable HNSCC were explored using small pieces of primary tumor and dissociated HNSCC cells after short-term culture. Viable cells after freezing were confirmed by adherence to tissue culture plates, cell morphology, and increased cell or colony density. Two cultures were immunostained for cytokeratin to confirm epithelial origin of viable cultured cells after freezing.. Six primary HNSCCs (two oral cavity, three larynx, one oropharynx) and two HNSCCs that had been passaged through a xenograft (two oral cavity) were dissociated to single cells and grown in short-term cell culture for 0 to 12 passages. After short-term culture, cells were frozen for up to 8 months, thawed, and replated. Frozen cells derived from all tumors (six primary and two xenografts) were successfully replated with cultures lasting >7 days with seven of eight tumors presenting increased colony or cell density over 1 week of growth after freezing. In total, 15 of 15 tested samples derived from six primary and two xenografted HNSCCs were viable after freezing.. In the current study, we show that biopreservation of primary or xenografted HNSCC using short-term cell culture is feasible. Initial short-term cell culture was required for successful storage and viability of frozen cells. These proof-of-principle studies, if more widely implemented, could improve preclinical testing of new therapies for HNSCC.

Topics: Carcinoma, Squamous Cell; Cryopreservation; Feasibility Studies; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Mouth Neoplasms; Oropharyngeal Neoplasms; Squamous Cell Carcinoma of Head and Neck; Tissue Banks; Transplantation, Heterologous; Tumor Cells, Cultured

Sarcomatoid basal cell carcinoma (BCC) is rare. In the literature, data on the prognosis of such a variant is somewhat contradictory. D2-40 immunoexpression has been shown to have prognostic connotations in carcinomas of organs other than the skin. However, although D2-40 immunoexpression has been investigated in "common" (nonsarcomatoid) BCC, it has not yet been studied in the spindle cell component of a sarcomatoid BCC. We present a sarcomatoid BCC on the neck of an 87-year-old man that has grown rapidly over the last few months. The sarcomatoid component of the tumor expressed several types of cytokeratins, such as AE1/AE3, CK 5/6, 34betaE12, and CAM 5.2. It was also positive for p63 and for D2-40 in a diffuse pattern.

Topics: Aged, 80 and over; Antibodies, Monoclonal, Murine-Derived; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Basal Cell; Gene Expression; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Metaplasia

Although down-regulation of GNG7 in cancer was reported before, its role in carcinogenesis is poorly understood. It belongs to a family of large G-proteins that may be involved in cell-contact-induced growth arrest and function in tumor suppression. In the present study, we stained immunohistochemically 188 tumors derived from larynx or floor of the mouth for GNG7 protein and confronted it with clinicopathologic data. Moreover, we performed bisulfite pyrosequencing to analyze GNG7 promoter methylation. We identified recurrent loss of GNG7 protein expression in 68/188 (36%) cases and promoter hypermethylation in (42/98; 43%) primary tumors, predominantly in young patients (p < 0.001). Loss of GNG7 expression correlated with hypermethylation of GNG7 promoter region (p < 0.001). Moreover, loss of GNG7 protein expression correlated with tumor size (p = 0.012) and lack of cervical metastasis (p = 0.02) whereas sustained expression correlated with keratinization (p = 0.008). Taken together, loss of GNG7 protein expression is a frequent event in head and neck cancer. Moreover, our data suggest that hypermethylation of the promoter region of GNG7 is probably the mechanism of the observed inactivation.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; DNA Methylation; DNA, Neoplasm; Female; Gene Expression Regulation, Neoplastic; GTP-Binding Protein gamma Subunits; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Promoter Regions, Genetic; Sequence Analysis, DNA; Tumor Burden; Uterine Cervical Neoplasms

Atypical fibroxanthoma is a cutaneous dermal malignancy that presents on the sun-damaged skin of elderly people. It requires a definitive diagnosis, from a high-grade sarcoma to a nonmesenchymal neoplasm. The recommended treatment protocol differs from similar histologically related tumors; thus, a diagnosis of atypical fibroxanthoma should fulfill strict histologic and immunohistochemical stain criteria. The use of these standards will exclude other skin malignancies, including malignant fibrous histiocytoma, angiosarcoma, malignant melanoma, and squamous cell carcinoma. This study was performed with the aim of identifying key immunostains to develop diagnostic criteria.. Forty-two cases were studied retrospectively over a 10-year period using a panel of immunostains.. The average age at presentation was 78 years, with a male predominance. The scalp was found to be the most common site of occurrence, although other investigators have found the forehead, cheeks, nose, and ears as the prevailing sites of presentation.. An extensive panel of immunohistochemical stains can be used to prove a diagnosis of atypical fibroxanthoma.

Topics: Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Diagnosis, Differential; Female; Head and Neck Neoplasms; Histiocytoma, Benign Fibrous; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplasm Proteins; Retrospective Studies; Skin Neoplasms; Vimentin

A high lactate content in malignant head and neck cancer (Head and neck squamous cell carcinomas, HNSCC) is associated with a higher risk of metastatic spread and lower overall patient survival. However, until present, the underlying mechanisms are not clearly understood. Here, a systematic comparison of glucose metabolism in HNSCC and homologous normal tissue is presented for the first time.. The concentrations of glucose, lactate and ATP were measured in cryobiopsies of 29 human HNSCC and of 9 normal mucosa using bioluminescence imaging. The protein expression of lactate dehydrogenase (LDH) was analyzed by Western blotting.. Tumors own a higher content of lactate and LDH in comparison with normal tissues. However, within the tumor group, the grade of LDH expression shows substantially strong variation and overlap with normal values. Furthermore, LDH expression was not correlated with tumor lactate content. Investigating a small subpopulation, patients with a short-term survival had significantly higher tumor lactate levels compared to patients with long-term survival.. The data provide clear evidence of an enhanced glycolysis in tumors compared to normal tissue. This may partially but not completely attributable to an elevated expression of LDH. High tumor lactate levels may be predictive for restricted patient survival. In conclusion, lactate measurements, for example non-invasively with MRT, should be advanced for use in clinical routine as a supportive tool for tumor diagnosis and prognosis.

Topics: Adenosine Triphosphate; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Blotting, Western; Carcinoma; Carcinoma, Squamous Cell; Female; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Gingiva; Glucose; Glycolysis; Head and Neck Neoplasms; Humans; Keratins; L-Lactate Dehydrogenase; Lactic Acid; Luminescence; Male; Middle Aged; Neoplasms, Squamous Cell; Predictive Value of Tests; Prognosis; Squamous Cell Carcinoma of Head and Neck

To evaluate the use of diffusion-weighted magnetic resonance imaging (DW-MRI) for nodal staging and its impact on radiotherapy (RT) planning.. Twenty-two patients with locally advanced head and neck squamous cell carcinoma underwent contrast-enhanced computed tomography (CT), as well as MRI (with routine and DW sequences) prior to neck dissection. After topographic correlation, lymph nodes were evaluated microscopically with prekeratin immunostaining. Pathology results were correlated with imaging findings and an RT planning study was performed for these surgically treated patients. One set of target volumes was based on conventional imaging only, and another set was based on the corresponding DW-MRI images. A third reference set was contoured based solely on pathology results.. A sensitivity of 89% and a specificity of 97% per lymph node were found for DW-MRI. Nodal staging agreement between imaging and pathology was significantly stronger for DW-MRI (kappa = 0.97; 95% confidence interval [CI], 0.84-1.00) than for conventional imaging (kappa = 0.56; 95% CI, 0.16-0.96; p = 0.019, by McNemar's test). For both imaging modalities, the absolute differences between RT volumes and those obtained by pathology were calculated. Using an exact paired Wilcoxon test, the observed difference was significantly larger for conventional imaging than for DW-MRI for nodal gross tumor volume (p = 0.0013), as well as for nodal clinical target volume (p = 0.0415) delineation.. These results suggest that DW-MRI is superior to conventional imaging for preradiotherapy nodal staging of head and neck squamous cell carcinoma, and provides a potential impact on organsparing and tumor control.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Diffusion Magnetic Resonance Imaging; Female; Head and Neck Neoplasms; Humans; Keratins; Lymphatic Metastasis; Male; Middle Aged; Neck Dissection; Neoplasm Staging; Prospective Studies; Protein Precursors; Radiotherapy Planning, Computer-Assisted; Sensitivity and Specificity; Tomography, X-Ray Computed

Cervical lymph node metastases from unknown primary sites account for approximately 3% to 9% of all head and neck malignant lesions. Squamous cell carcinoma is the most common type of cervical metastatic carcinoma. Our aim was to investigate the possibility of determining the site of primary tumors using an immunohistochemical diagnostic panel in metastatic cervical lymph nodes. Expression profiles of cytokeratins, 5/6; 8/18; 10; 13; 14; and 19, p16, and pRb were evaluated in 101 consecutive patients with cervical nodal metastasis who had undergone neck dissection to treat known head and neck squamous cell carcinoma (primary sites: 16, oral cavity; 38, oropharynx; 26, hypopharynx; 21, larynx). Cytokeratin 10 was more frequently expressed in oral cavity primary tumors, whereas cytokeratin 19 staining was more frequently observed in tumors originated from the pharynx and larynx. The expression of p16 and altered pRb status (0% or >50%) were more frequently observed in oropharynx primary tumors. To select the best subset among the 8 antibodies tested, classification and regression tree analysis was performed. The analysis correctly classified the four primary sites (25.0% of oral cavity, 89.5% of oropharynx, 30.8% of hypopharynx, and 57.1% of larynx) using 5 variables (histologic subtype, p16, cytokeratins 10 and 19, and pRb). The p16 was the single best predictor. The classification tree method using immunostaining profiles of p16, cytokeratins 10 and 19, or pRb may be helpful in the identification of the primary site of metastatic squamous cell carcinoma with occult primary.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Chi-Square Distribution; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasms, Unknown Primary; Retrospective Studies; Tissue Array Analysis

to present and discuss a high-performance negative depletion method for the isolation of circulating tumor cells (CTCs) in the blood of patients with head and neck cancer and to determine the correlation between the presence of CTCs and early clinical outcome in these patients.. prospective clinical follow-up study of patients with squamous cell carcinoma of the head and neck (SCCHN) undergoing surgical intervention, who had peripheral blood examined for the presence of CTCs.. the study population comprised 48 patients diagnosed as having SCCHN and undergoing surgical intervention.. a negative depletion process to isolate and quantify CTCs from the blood of patients with SCCHN using immunomagnetic separation was developed and validated. Immunostaining for cytokeratin was performed on the enriched samples to determine the number of CTCs extracted from each patient's blood sample. Correlation of the presence of CTCs, tumor stage, nodal status, clinical characteristics, and outcome was made.. disease-free survival.. our initial data, that have a mean follow-up of 19.0 months, suggest that patients with no detectable CTCs per milliliter of blood had a significantly higher probability of disease-free survival (P = .01). There was no correlation between the presence of CTCs with regard to age, sex, tumor site, stage, or nodal involvement.. our enrichment technology, based on the removal of normal cells, has been used on the peripheral blood of patients with head and neck cancer for which follow-up data were collected. If no CTCs were present, a statistically significant improved disease-free survival was observed in SCCHN. A blood test with such a prognostic capability could have important implications in the treatment of patients with head and neck cancer.

Topics: Carcinoma, Squamous Cell; Disease-Free Survival; Female; Follow-Up Studies; Head and Neck Neoplasms; Humans; Immunohistochemistry; Immunomagnetic Separation; Keratins; Male; Middle Aged; Neoplasm Staging; Neoplastic Cells, Circulating; Prognosis; Prospective Studies

Topics: Aged, 80 and over; Biopsy, Fine-Needle; Cytological Techniques; Female; Head and Neck Neoplasms; Humans; Keratins; Lymph Nodes; Lymphoma, Non-Hodgkin

We investigated p16(INK4A) expression in branchial cleft cysts and its utility in distinguishing branchial cleft cysts from metastatic head and neck squamous cell carcinomas (SCCs) in fine-needle aspiration biopsies (FNABs).. A study set comprising 41 resections (15 SCC and 26 branchial cleft cysts) and a test set of 15 FNABs (11 SCC and 4 branchial cleft cysts) were analyzed with p16(INK4A) immunohistochemistry and human papillomavirus (HPV) polymerase chain reaction (PCR)/pyrosequencing. Cases with discrepant p16(INK4A) and PCR/pyrosequencing results were further evaluated with HPV in situ hybridization (ISH). SCCs were divided into keratinizing SCC and nonkeratinizing SCC groups and site of origin.. Metastatic oropharyngeal nonkeratinizing SCC in the study set exhibited diffuse, strong p16(INK4A) (7 of 7) and HPV16 DNA positivity (6 of 6), while keratinizing SCC from the larynx and oral cavity was negative for p16(INK4A). p16(INK4A) reactivity in the branchial cleft cyst study set was characterized by focal, strong staining (6 of 21) involving the superficial squamous epithelium. HPV DNA was identified in 7 of 19 branchial cleft cyst study set cases by PCR/pyrosequencing, but these cases were negative by HPV ISH. In the test set, oropharyngeal nonkeratinizing SCC exhibited diffuse, strong p16(INK4A) (3 of 3) and HPV16 DNA (2 of 2), while metastatic keratinizing SCC was negative for p16(INK4A) and HPV DNA. All 4 FNABs of branchial cleft cysts were negative for p16(INK4A). Diffuse, strong p16(INK4A) correlated with oropharyngeal origin (P=.001) and nonkeratinizing morphology (P=.0001).. Branchial cleft cysts can exhibit focal strong reactivity limited to the superficial squamous epithelium and glandular epithelium. Although p16(INK4A) immunohistochemistry may be helpful in distinguishing oropharyngeal nonkeratinizing SCC from branchial cleft cysts in FNAB specimens, it is not helpful in cases of keratinizing SCC because these cases are typically negative for p16(INK4A).

Topics: Adolescent; Aged; Biopsy, Fine-Needle; Branchioma; Carcinoma, Squamous Cell; Cyclin-Dependent Kinase Inhibitor p16; Cysts; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Oropharynx

A great deal of attention has been given to epithelial differentiation in Ewing family of tumors (EFTs) in recent years, with studies showing variable keratin expression in 20% to 32% of cases. A 29-year-old man presented with a right-sided Horner syndrome suggesting a sympathetic chain related lesion. No radiologic evidence of a mass was appreciated initially. Several months later, he developed right vocal cord palsy and a large right-sided neck mass. An open biopsy demonstrated a high-grade malignant neoplasm with sheets of undifferentiated round cells infiltrating soft tissues and a large peripheral branch of the vagus nerve. Focally, the tumor abruptly produced keratinizing cells and frank squamous pearls. The tumor showed diffuse expression of CD99, high molecular weight keratin, p63, cytokeratin (CK) 5/6, AE1/AE3, CAM5.2, CK19, and focal CK14. It was negative for muscle-specific actin, desmin, MyoD1, MYF-4, S100, and CK7. Ultrastructurally, abundant cytoplasmic tonofilaments and well-formed desmosomes were demonstrated. Initial diagnosis was a metastatic squamous cell carcinoma of probable upper aerodigestive origin. Subsequently, the tumor was shown to harbor the t(11;22) involving EWSR1 and FLI-1 by reverse transcription-polymerase chain reaction, characteristic of EFT's, which was confirmed by dual color break apart fluorescence in-situ hybridization analysis. This tumor is related to, if not an example of the recently described "adamantinoma-like" EFT and demonstrates a potential diagnostic pitfall. It seems to be the first EFT to arise within, or in close proximity to the cervical sympathetic chain of ganglia. It is also the first EFT with complex epithelial differentiation arising outside the extremities and with diffuse expression of high molecular weight keratin and p63.

Topics: Adult; Biomarkers, Tumor; Epithelium; Head and Neck Neoplasms; Horner Syndrome; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratins; Male; Membrane Proteins; Oncogene Proteins, Fusion; Proto-Oncogene Protein c-fli-1; Reverse Transcriptase Polymerase Chain Reaction; RNA-Binding Protein EWS; Sarcoma, Ewing; Translocation, Genetic; Vagus Nerve

At later stages of tumor progression, epithelial carcinogenesis is associated with transition to a mesenchymal phenotype, which may contribute to the more aggressive properties of cancer cells and may be stimulated by growth factors such as epidermal growth factor and transforming growth factor-beta. Previously, we found that cells derived from a nodal metastatic squamous cell carcinoma are highly proliferative and motile in vitro and tumorigenic in vivo. In the current study, we have investigated the role of vimentin in proliferation and motility. Cells derived from nodal metastasis express high levels of vimentin, which is undetectable in tumor cells derived from a synchronous primary lesion of tongue. Vimentin expression was enhanced by epidermal growth factor and transforming growth factor-beta both independently and in combination. Use of RNA interference resulted in the generation of stable cell lines that express constitutively low levels of vimentin. RNA interference-mediated vimentin knockdown reduced cellular proliferation, migration, and invasion through a basement membrane substitute by 3-fold compared with nontargeting controls. In addition, cells with reduced vimentin reexpressed differentiation-specific keratins K13, K14, and K15 as a result of increased gene transcription as judged by quantitative PCR and promoter-reporter assays. Furthermore, cells in which vimentin expression was reduced showed a greatly decreased tumorigenic potential, as tumors developing from these cells were 70% smaller than those from control cells. The data suggest that reversal of the mesenchymal phenotype by inhibiting vimentin expression results in reexpression of epithelial characteristics and reduced tumor aggressiveness.

Topics: Animals; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line, Tumor; Cell Movement; Cell Proliferation; Down-Regulation; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Intercellular Signaling Peptides and Proteins; Keratins; Mice; Mice, Nude; Neoplasm Metastasis; RNA Interference; RNA, Messenger; RNA, Small Interfering; Vimentin

High-grade neuroendocrine carcinomas of the head and neck overlap significantly in morphology with both basaloid squamous and solid-type adenoid cystic carcinomas. High-grade neuroendocrine carcinomas have sheets of small cells with scant cytoplasm, granular chromatin, and inconspicuous nucleoli. Basaloid squamous and adenoid cystic carcinomas are aggressive variants of their respective tumor types which both have nests of basaloid tumor cells with round nuclei, little cytoplasm, and inconspicuous nucleoli. As the management and prognosis of these tumors are very different, it is important to differentiate them. We performed high molecular weight cytokeratin (CK) and p63 immunohistochemistry on 19 neuroendocrine carcinomas, 18 basaloid squamous carcinomas, and 11 solid-type adenoid cystic carcinomas. All tumors were immunostained for p63, CK 34betaE12, CK 5/6, synaptophysin, chromogranin-A, S-100, and smooth muscle actin. All basaloid squamous and adenoid cystic carcinomas were positive for CK 5/6 and 34betaE12. Only 4 and 5 of the 19 neuroendocrine carcinomas, respectively, were positive for these markers. Staining was focal in the neuroendocrine cases when positive, whereas almost all basaloid squamous and adenoid cystic carcinomas showed strong staining. Almost all tumors of each type were positive for p63, including neuroendocrine carcinomas, but with different staining patterns. Basaloid squamous carcinomas were diffusely positive, neuroendocrine carcinomas were diffusely positive, but with weak staining, and adenoid cystic carcinomas showed a distinct pattern with staining at the periphery of the cell nests only. We conclude that high molecular weight cytokeratin immunostaining is helpful in distinguishing high-grade neuroendocrine carcinomas from similar tumor types.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Basosquamous; Carcinoma, Neuroendocrine; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratin-5; Keratin-6; Keratins; Male; Membrane Proteins; Middle Aged; Molecular Weight

An unusual tumor of the neck in a 56-year-old female is reported. The tumor was composed of tubules and small cords of epithelial cells dispersed in the fibromyxoid and adipose stroma. At the periphery, the tumor was not encapsulated and its border was intermingled with the subcutaneous fat. Lack of decapitation secretion and the absence of long tubules suggested an eccrine origin; however, in some of the tumor areas, the cells showed brightly eosinophilic copious cytoplasm that may indicate an apocrine differentiation. As an area of chondroid metaplasia was identified, the diagnosis of a mixed tumor was rendered. This unusual type of skin adnexal neoplasm with unique relation of the epithelial component to the surrounding adipose tissue requires differentiation with the primary cutaneous and metastatic carcinomas.

Topics: Adipocytes; Adipose Tissue; Biomarkers, Tumor; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lipoma; Middle Aged; Neoplasm Metastasis; Neoplasms, Complex and Mixed; Skin Neoplasms

Spindle cell carcinoma (SpCC) is a rare microscopic type of cancer of the mouth and oropharynx. Although SpCC is thought to arise from squamous cell carcinoma (SCC), it carries a worse prognosis.. To find out the difference in immunohistochemical expression of cytokeratin, vimentin and smooth-muscle actin, and mutational alterations in the K-ras oncogene between the two tumours, in an attempt to characterise SpCC.. Immunohistochemical analysis was performed by standard avidin-biotin complex method in 35 cases each of SpCCs and SCCs. DNA extracted from paraffin wax-embedded tumours was used for PCR followed by single-strand conformation polymorphism for mutational analysis of K-ras exon 1 and exon 2.. In the SpCC group, cytokeratin positivity was significantly higher in epithelial areas (52.2%) than in spindle cell areas (16.1%), whereas vimentin was more positive in spindle cell areas (18.7%) than epithelial areas (2.7%). Cells intermediate between epithelial and spindle cell areas were consistently positive for both cytokeratin and vimentin. Cytokeratin was found to be significantly more positive in SCC (72.6%) than the squamous component and spindle cell component of SpCC. In this study, no mutation was detected in the K-ras gene of either the SpCC or SCC group.. The spindle cell component of SpCC is intermixed with cells that are morphologically mesenchymal but express dual antigen-positivity characteristic of epithelial (cytokeratin) and mesenchymal (vimentin) cells. These, possibly, are cells in transition suggesting that SpCC may be a sarcomatous metaplasia of SCC.

Topics: Actins; Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; DNA Mutational Analysis; DNA, Neoplasm; Female; Genes, ras; Genetic Predisposition to Disease; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Neoplasm Proteins; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Prospective Studies; Vimentin

Merkel cells carcinoma (MCC) is an uncommon skin lesion, considered a malignancy of the neuroendocrine system, which is found mainly in elderly people. Its incidence is highly correlated with sun exposure or immunodeficiency syndromes. MCC is often an aggressive tumour with high tendency for local recurrence, lymph node involvement and distant metastasis. To our best knowledge 20 cases originated from the auricle have been described, 2 of them arising from external ear canal. The authors report a case of the ear canal characterized by two others synchronous tumours and the occurrence of a malignant high grade lymphoma, in which contribute of the pathologist was essential for a critical review. MCC diagnosis is not always easy for its pathological and clinical features and it should always be considered in presence of lymphoma. A multidisciplinary approach is basic.

Topics: Aged; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Biopsy, Fine-Needle; Carcinoma, Merkel Cell; Ear Neoplasms; Ear, External; Fatal Outcome; Head and Neck Neoplasms; Humans; Keratins; Lymphoma, Follicular; Magnetic Resonance Imaging; Male; Neoplasms, Second Primary; Parotid Gland; Skin Neoplasms

Topics: Aged; Biomarkers, Tumor; Facial Neoplasms; False Positive Reactions; Head and Neck Neoplasms; Histiocytoma, Benign Fibrous; Humans; Keratins; Male; Neoplasm Proteins; Scalp; Skin Neoplasms

To investigate the use of immunohistochemistry in distinguishing necrotizing sialometaplasia (NSM) from squamous cell (SCC) and mucoepidermoid carcinoma (MEC) by (i) the identification of myoepithelial cells and (ii) cytokeratin (CK) expression.. Thirteen cases with the histological changes of NSM, eight SCCs and eight MECs were examined with the following immunohistochemical markers: calponin, S100, smooth muscle actin (SMA), p63, CK7, CK5, CK6 and CAM5.2. The distribution and intensity of staining were recorded. Residual myoepithelial cells (best demonstrated by calponin and SMA) were identified at the periphery of the epithelial islands in all cases of NSM (although not in all islands), in contrast to MEC and SCC. S100 showed a similar pattern, although staining fewer cells. Moderate rather than extensive expression of CK7 may help to distinguish NSM from MEC.. Identification of myoepithelial cells and CK7 expression may help to distinguish NSM from its mimics.

Topics: Actins; Biomarkers; Biomarkers, Tumor; Calcium-Binding Proteins; Calponins; Carcinoma, Mucoepidermoid; Carcinoma, Squamous Cell; Diagnosis, Differential; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Keratins, Type II; Membrane Proteins; Microfilament Proteins; S100 Proteins; Sialometaplasia, Necrotizing

The human p63 gene codes for multiple protein isoforms and is commonly over-expressed in squamous cell carcinoma of head and neck (SCCHN). This expression is predominantly of the DeltaN- and beta-isoforms, the former lacking the p53-related transactivation domain. p63 can activate or repress transcription of p53 and p73 target genes, but also has unique transcriptional targets and, unlike other p53 family members, is required for normal development and differentiation of squamous epithelia. We have identified novel targets of p63, using microarray analysis of SCCHN cells that stably over-express individual DeltaNp63 isoforms. All three isoforms induced expression of the cancer stem cell marker, CD44, with the DeltaNp63beta isoform showing strongest induction. Using chromatin immunoprecipitation, we were unable to show direct binding of p63 to the CD44 promoter, but found that p63 specifically increased expression of CD44 lacking variant exon 2. Each of the DeltaNp63 isoforms up-regulated expression of keratins 6A and 14 and down-regulated expression of keratins 4 and 19, in keeping with their expression patterns in SCCHN. The data strengthen the idea that p63 has key roles in regulating normal and abnormal differentiation processes through both induction and repression of genes with opposite functions. The identification of up-regulation and differential splicing of CD44 following p63 over-expression indicates roles in the regulation of adhesion, metastasis and the cancer stem cell phenotype.

Topics: Carcinoma, Squamous Cell; DNA-Binding Proteins; Down-Regulation; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Hyaluronan Receptors; Keratins; Mouth Mucosa; Neoplasm Proteins; Oligonucleotide Array Sequence Analysis; Protein Isoforms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Trans-Activators; Transcription Factors; Tumor Cells, Cultured; Tumor Suppressor Proteins; Up-Regulation

To explore possible range of gene expression profiles in head and neck squamous cell carcinomas (HNSCC) and pairwised normal controls from Sudanese (n = 72) and Norwegian (n = 45) patients using a 15K cDNA microarray and to correlate the findings with clinicopathologic variables.. Samples from Sudan were grouped according to anatomic location/patients' habit of toombak (snuff) use, and 37 pools of 2 to 11 tumors matched to 37 pools of their normal controls from the same patients, respectively, were prepared. For Norway, eight pools of 3 to 11 tumors matched to eight pools of their normal controls from the same patients, respectively, were prepared according to anatomic location. Pools (n = 45) were hybridized to microarrays. For controls, 33 of the pools were hybridized against Human Reference RNA. Scanned array images were recorded, and data analysis was done in groups. For verification, results for selected genes were analyzed using quantitative real-time PCR/immunohistochemistry.. We identified 136 genes from Sudan and 154 from Norway as differentially expressed between tumors and controls. Changes of the genes found were confirmed in >70% of the pools by hybridization against Reference RNA. Seventy-three genes and three main pathways (signal transduction, cell communication, and ligand-receptor interaction) were of relevance to the HNSCCs from both countries. Hierarchical clustering of the 73 genes identified subclasses of mixed tumors from the two populations, two independent subgroups for Norwegian tumors by their anatomic sites, and five subgroups for Sudanese tumors by their toombak habits. Quantitative real-time PCR/immunohistochemistry validated the microarray-based data.. Differences in gene expression between tumor and nontumor tissues were identified in HNSCCs. Analysis of the two population groups revealed a common set of 73 genes within three main biological pathways. This indicates that the development of HNSCCs is mediated by similar biological pathways regardless of differences related to race, ethnicity, lifestyle, and/or exposure to environmental carcinogens. Of particular interest, however, was the valuable association of gene expression signature found with toombak use and anatomic site of the tumors.

Topics: Aged; Antigens, Nuclear; Black People; Carcinoma, Squamous Cell; Chemotactic Factors; Cluster Analysis; DNA-Binding Proteins; Female; Fibronectins; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Ku Autoantigen; Life Style; Male; Middle Aged; Norway; RNA, Messenger; S100 Proteins; Sudan; White People

Screening of head and neck carcinoma patients with the proteomics-based AMIDA technology yielded a set of tumour-associated antigens, including the intermediate filament protein cytokeratin 8 (CK8). The expression pattern and specificity of CK8 was compared with those of the established markers pan-cytokeratins and CK13, and with that of the proliferation marker Ki67. Expression of CK8 correlated positively with malignancies of the head and neck areas. CK8 was not expressed in healthy epithelium, except for some rare cases of cells of the basal layer and laryngeal tissue. In contrast, the vast majority of head and neck squamous cell carcinomas and metastases strongly expressed CK8. Interestingly, CK8 de novo expression correlated with dysplastic areas of oral leukoplakic lesions, while hyperplastic leukoplakia remained CK8-negative but strongly panCK and CK13 positive. Thus, CK8 is an attractive marker molecule for a differentiated diagnosis of leukoplakia and head and neck carcinomas, which possesses notedly improved specificity as compared with panCK and CK13.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Leukoplakia, Oral

A 64 year old man presented with progressive impairment of right sided cranial nerves. Chronic immunosuppression for renal transplantation had resulted in multiple squamous cell carcinomata of the head and neck. Magnetic resonance imaging and subsequent right facial nerve biopsy confirmed perineural spread of a squamous cell carcinoma as the cause of the multiple cranial neuropathies.

Topics: Carcinoma, Squamous Cell; Facial Nerve Diseases; Head and Neck Neoplasms; Humans; Keratins; Magnetic Resonance Imaging; Male; Middle Aged

The aim of the present study was to evaluate the occult lymph node carcinomatous diffusion in head and neck squamous cell carcinoma (HNSCC). A total of 1328 lymph nodes from 31 patients treated between 2004 and 2005 were prospectively evaluated by routine haematoxylin-eosin-safran (HES) staining, immunohistochemistry (IHC) and real-time Taqman reverse-transcriptase polymerase chain reaction (real-time RT-PCR) assay. Amplification of cytokeratin 19 (CK19) mRNA transcripts using real-time RT-PCR was used to quantify cervical micrometastatic burden. The cervical lymph node metastatic rates determined by routine HES staining and real-time RT-PCR assay were 16.3 and 36.0%, respectively (P<0.0001). A potential change in the nodal status was observed in 13 (42.0%) of the 31 patients and an atypical pattern of lymphatic spread was identified in four patients (12.9%). Moreover, CK19 mRNA expression values in histologically positive lymph nodes were significantly higher than those observed in histologically negative lymph nodes (P<0.0001). These results indicate that real-time RT-PCR assay for the detection of CK19 mRNA is a sensitive and reliable method for the detection of carcinomatous cells in lymph nodes. This type of method could be used to reassess lymph node status according to occult lymphatic spread in patients with HNSCC.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Prospective Studies; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity

Immunodetection of GLUT1, p63 and phospho-histone H1 in invasive head and neck squamous carcinoma: correlation of immunohistochemical staining patterns with keratinizationAims : To examine invasive head and neck squamous carcinomas for expression of GLUT1, a glucose transporter and marker of increased glucose uptake, glycolytic metabolism and response to tissue hypoxia; p63, a p53 homologue that is a marker of the undifferentiated proliferative basaloid phenotype; and phospho-histone H1, a marker of activation of the cell cycle-promoting cyclin-dependent kinases 1 and 2. Methods : Routinely processed slides from 34 invasive squamous carcinomas, including 25 with intraepithelial components, were immunostained with anti-GLUT1 (Chemicon), anti-p63 (4A4, Santa Cruz), and antiphospho-histone H1 (monoclonal 12D11). Results : In keratinizing carcinomas, all three markers were most commonly immunodetected peripherally, with loss of expression in central keratinized zones. In contrast, in non-keratinizing carcinomas, p63 and phospho-histone H1 expression was most commonly observed throughout tumour nests and anti-GLUT1 stained in a pattern suggestive of hypoxia-induced expression ('antistromal' staining), in which cells at the tumour-stromal interface were GLUT1- and cells in central, perinecrotic zones showed progressive induction of GLUT1. Intraepithelial components also displayed basal and 'antibasal' GLUT1 staining patterns, homologous to the pro- and antistromal patterns in invasive carcinoma; basal patterns in intraepithelial lesions appeared to be more predictive of keratinizing invasive carcinoma and antibasal intraepithelial staining more predictive of non-keratinizing poorly differentiated carcinomas. Conclusions : Keratinizing and non-keratinizing squamous carcinomas differ in expression patterns of GLUT1, p63 and phospho-histone H1. In the former, all three markers were typically suppressed in conjunction with keratinization; in the latter, GLUT1 expression was more likely to occur in a hypoxia-inducible pattern and expression of p63 and phospho-histone H1 was unsuppressed. GLUT1 expression patterns in intraepithelial lesions may be predictive of the differentiation status of the associated invasive carcinoma.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease Progression; DNA-Binding Proteins; Glucose Transporter Type 1; Head and Neck Neoplasms; Histones; Humans; Immunohistochemistry; Keratins; Neoplasm Invasiveness; Phosphoproteins; Phosphorylation; Trans-Activators; Transcription Factors; Tumor Suppressor Proteins

We experienced two cases of cutaneous dermoid cysts (DC). To elucidate the histogenesis of DC, we have studied cytokeratin (CK) expression in DC using ten different anti-keratin antibodies against CK1, 7, 8, 10, 14, 15, 16, 17, 18 and 19, and anti-filaggrin (filament aggregating protein) antibody. In the cyst wall of DC, CK1 and 10 were expressed in suprabasal layer, and CK14 was limited to the basal layer. In sebaceous gland-like structures, CK14 was detected in sebaceous acinus, and CK17 was detected in sebaceous duct. The other CKs were not detected. Filaggrin was intensely detected in the granular layer in the cyst wall of DC. CK expression profile of DC was similar to follicular infundibulum and sebaceous gland. These results suggested that DC differentiates towards follicular infundibulum and mature sebaceous gland.

Topics: Adult; Antibodies; Cell Differentiation; Dermoid Cyst; Female; Filaggrin Proteins; Head and Neck Neoplasms; Humans; Intermediate Filament Proteins; Keratins; Male; Middle Aged; Sebaceous Glands; Skin Neoplasms

To analyze the lymphatic distribution of metastatic carcinomatous cells in cervical lymph nodes in head and neck squamous cell carcinoma (HNSCC).. We retrospectively reviewed 119 patients treated in our hospital for HNSCC (1999-2004). Topography of the neck dissection specimens was prospectively classified according to the classification of Robbins. The 4000 lymph nodes were analyzed by optical microscopy using hematoxylin-eosin-safran (HES) staining. In cases of negative results in level II, cytokeratin (AE1/AE3) immunodetection was performed.. Metastases were visualized using HES in 6.4% of lymph nodes for oral cavity, and 4.7% of oropharyngeal, 4.4% of hypopharyngeal, and 1.3% of endolaryngeal cancers. The highest incidence of nodal metastasis was observed in level IIa (P < 0.01). In eight patients (6.7%) with lymph node metastases, level II was spared. In these patients, all 134 nodes histologically negative on HES were confirmed to be negative by IHC.. Level IIa is the main level involved in regional metastases of HNSCC, regardless of the primary site of cancer. However, in eight (6.7%) patients, level II was spared, as confirmed by IHC. In these cases, level II did not represent the first step of drainage from the tumor. The sentinel lymph node technique in HNSCC is discussed in light of these results.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Coloring Agents; Female; Fluorescent Dyes; Head and Neck Neoplasms; Humans; Keratins; Laryngeal Neoplasms; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neck; Neck Dissection; Neoplasm Staging; Pharyngeal Neoplasms; Prospective Studies; Retrospective Studies; Sentinel Lymph Node Biopsy

The identification of a reliable circulating tumor marker in squamous cell carcinoma of the head and neck (SCCHN) could assist in diagnosis and monitoring response to therapy.. The aim of this study was to investigate the effectiveness of the serum tumor markers CYFRA 21-1, TPA-M, SCCA, and CEA.. Serum levels of CYFRA 21-1, TPA-M, SCCA, and CEA were measured in 136 patients with a histologically proven SCCHN before and after treatment and in 125 healthy subjects, as controls. We evaluated the sensitivity and specificity of these tumor markers and to correlate their levels with tumor staging, grading, or performance status.. The study showed that none of the above markers presented satisfactory specificity and sensitivity in early diagnosis. In comparison with the other markers, TPA-M was the most effective of all markers and indicated a positive correlation with the grade of differentiation and nodal status. A remarkable correlation between high levels of TPA-M and CYFRA 21-1 in advanced stages (III, IV) of cancer has been shown.. All the tumor markers that were studied have significant limitations in the early diagnosis of cancer, but TPA-M and CYFRA 21-1 may have a role in monitoring the success of therapy and follow up of patients with SCCHN.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Male; Middle Aged; Neoplasm Staging; Prospective Studies; Sensitivity and Specificity; Serpins; Tissue Polypeptide Antigen

We investigated the relationship between cell growth and differentiation and COX-2 expression in oral squamous cell carcinoma (SCC) in vitro and in vivo. Treatment of SCC25 oral squamous carcinoma cells with sodium butyrate (SB) at 0.5-5 mM or all-trans retinoic acid (ATRA) at 3-300 microM inhibited cell growth and induced apoptosis in a dose-dependent manner with concomittant increases in expression of keratin 13, p21WAF1/Cip1 and p27Kip1 and decreases in expression of COX-2. These effects were more pronounced with SB than with ATRA. Injection of SB or ATRA near SCC25-derived tumors in nude mice resulted in inhibition of growth and elevation of differentiation of the tumor accompanied by marked keratinization and increased expression of keratin 13 and decreased expression of COX-2. These results show that the differentiation-inducing agents, particularly SB, suppress growth of oral squamous carcinoma cells through apoptosis and induce cell differentiation possibly through mechanisms involving COX-2, p27Kip1 and/or p21WAF1/Cip1 in vitro and in vivo.

Topics: Animals; Apoptosis; Blotting, Western; Butyrates; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Cyclooxygenase 2; DNA Primers; Female; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Isobutyrates; Keratins; Membrane Proteins; Mice; Mice, Inbred BALB C; Mice, Nude; Mouth Neoplasms; Prostaglandin-Endoperoxide Synthases; Reverse Transcriptase Polymerase Chain Reaction; RNA; Time Factors; Tretinoin; Tumor Suppressor Proteins

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

Although head and neck cancer is a common malignancy, investigations have not yet improved the poor prognosis of patients. Therefore, it is important to find new cancer treatment modalities. Recent studies showed that cyclooxygenase, especially its isoform cyclooxygenase-2, is involved in tumorigenesis, tumor growth and metastasis. Inhibition of this enzyme by cyclooxygenase inhibitors has been shown to be antiproliferative in numerous cancer cell lines. The aim of this study was to investigate if the selective cyclooxygenase-2 inhibitor nimesulide could enhance cytotoxicity of the standard chemotherapeutic agent cisplatin. Head and neck squamous cell cancer cells were incubated with nimesulide and/or cisplatin, and counted after 24, 48 and 72 h treatment. Visualization of apoptotic cells was done by immunohistochemistry. We demonstrated that nimesulide inhibits the cytotoxic effect of cisplatin in HNSCC cells. Therefore, COX-2 inhibitor nimesulide may not be a good partner for cisplatin in combination therapy for cancer.

Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cisplatin; Cyclooxygenase 2; Cyclooxygenase Inhibitors; Dose-Response Relationship, Drug; Drug Antagonism; Drug Synergism; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Sulfonamides; Time Factors

We performed a prospective study to determine the cutoff value and the prognostic value of Cyfra 21-1, a serum tumor marker, in head and neck squamous cell carcinoma (HNSCC).. The serum concentration of Cyfra 21-1 was measured in a group of 300 patients (group 1) with HNSCC, in a control group of 71 healthy subjects (group 2), and in a group of 73 patients with a nonmalignant tumor or inflammatory disease (group 3). The concentrations were compared between the various groups and subgroups; the cutoff value was calculated with a receiver operating characteristic curve. Furthermore, the serum concentrations of Cyfra 21-1 before treatment in the group of 300 patients were compared with the stage of the disease and with the evolution of the overall survival rate and the disease-free survival rate. Finally, to determine whether Cyfra 21-1 is an independent prognostic factor, we compared the concentrations, by a Cox model, with the classic prognostic factors of HNSCC.. At the cutoff value of 1 ng/mL, the specificity was 94% and the sensitivity was 72%. The serum concentrations of Cyfra 21-1 were statistically correlated with the stage of the disease. The overall survival rate and the disease-free survival rate were lower in patients with high serum concentrations, and these differences were statistically significant (p < .001). The Cox model allows us to conclude that Cyfra 21-1 is a prognostic marker that is independent of other classic prognostic factors.. Cyfra 21-1 is an interesting tumor marker that could be proposed for the early detection of HNSCC with a cutoff value of 1 ng/mL. Furthermore, Cyfra 21-1 can be considered an independent prognostic marker.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease-Free Survival; Female; Head and Neck Neoplasms; Humans; Hypopharyngeal Neoplasms; Keratin-19; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Multivariate Analysis; Oropharyngeal Neoplasms; Prognosis; ROC Curve; Sensitivity and Specificity

We report the case of an 85-year-old man who presented with an enlarging tumor on the right temple. Histologically, spindle-shaped cells were proliferating continuously within well-differentiated squamous cell carcinoma that was immunohistochemically negative for vimentin and alpha-smooth muscle actin (ASMA) but positive for cytokeratin (AE1/3). These spindle-shaped cells expressed vimentin and ASMA but not cytokeratin (AE1/3). Intermediate cells with round nuclei and small amounts of cytoplasm displayed slight expression of vimentin, ASMA, and cytokeratin. This case illustrated spindle cell squamous cell carcinoma with mesenchymal metaplasia of malignant epithelial cells.

Topics: Actins; Aged, 80 and over; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Mesoderm; Skin Neoplasms; Vimentin

Topics: Cheek; Diagnosis, Differential; Head and Neck Neoplasms; Humans; Immunophenotyping; Keratins; Sense Organs

To report three cases of primary carcinoma of the neck arising in multilocular thymic cysts (MTC).. The patients were three men aged 47, 50 and 52 years who presented with a painless neck mass of several weeks' duration. The patients had no history of previous surgical procedures or of malignancy elsewhere. The tumours in all three patients were located on the right lateral side of the neck; all patients underwent complete surgical resection of the mass. Grossly, the tumours were cystic and measured between 20 and 30 mm in greatest diameter. Histologically, the tumours showed cyst walls lined by squamous epithelium. The cyst walls contained prominent germinal centres with lymphoid hyperplasia, cholesterol cleft granulomas, and scattered keratinized structures reminiscent of Hassall's corpuscles. In addition, a neoplastic cellular proliferation composed of round to oval cells arranged in sheets and originating from the lining of the cystic structures was present. The neoplastic cells showed moderate amounts of eosinophilic cytoplasm, round nuclei, and, in some areas, prominent nucleoli. Mitotic figures were easily found, and cellular pleomorphism was present in several areas. In two cases the tumours showed features of basaloid carcinoma of the thymus, while in one case the pattern was that of squamous cell carcinoma. Immunohistochemical studies for keratin showed a strong positive reaction in the tumour cells, while leucocyte common antigen strongly stained the lymphoid background. Follow-up information obtained in two patients showed them to be alive 6 months after initial diagnosis. One patient was lost to follow-up.. The cases described here represent an unusual variant of carcinoma arising in multilocular thymic cyst in the neck region.

Topics: Adenocarcinoma; Biomarkers, Tumor; Disease-Free Survival; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Mediastinal Cyst; Middle Aged

The presence or absence of metastatic disease in cervical lymph nodes is the single most important determinant of therapy and prognosis for patients with head and neck squamous cell carcinoma (HNSCC). However, histologic examination fails to detect metastatic disease in a subset of neck dissection specimens. The accuracy of neck staging may be improved by the use of molecular techniques. Cytokeratins 5, 14, and 20 may be appropriate markers for HNSCC because they are expressed in HNSCC but not in lymphatic tissue.. To test the sensitivity of detection of cytokeratin 5, 14, and 20 messenger RNA by quantitative reverse transcription polymerase chain reaction (RT-PCR), full-length coding DNA sequences were cloned and transcribed. The expression of cytokeratin 5, 14, and 20 messenger RNA was quantified in 4 HNSCC cell lines and 11 tumors. A cell culture lymph node model was created.. As few as 32 molecules of cytokeratin 14 could be detected using quantitative RT-PCR. Cytokeratins 5 and 14 were easily detected in all 4 HNSCC cell lines and almost all tumors. Cytokeratin 20 was not a useful marker, as expression was absent or significantly reduced in cell lines and tumors. In the lymph node model, cytokeratin 14 quantitative RT-PCR was able to detect 1 cancer cell in a background of 10 million lymphatic cells.. Quantitative RT-PCR detection of cytokeratin 5 or 14 is a sensitive new molecular technique that may be used for detection of cervical micrometastases in head and neck cancer.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Keratins; Lymphatic Metastasis; Molecular Probes; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Tumor Cells, Cultured

Head and neck squamous cell cancer (HNSCC) requires precise characterization of their biological properties to better stratify treatment approaches. Some biological features of tumor cells are related to altered glycosylation of their surface. This study characterized alpha2,3/6-N-acetylneuraminic acid (NeuNAc) expression in adult squamous epithelia of primary and metastatic HNSCC in relation to expression of well established differentiation markers, such as cytokeratins. The expression of NeuNAc was assessed by plant lectins: Maackia amurensis agglutinin type 2 (MAA) and Sambucus nigra agglutinin (SNA) specific for alpha2,3NeuNAc and alpha2,6NeuNAc, respectively. Double labeling studies at the single-cell level were performed to compare alpha2,3/6NeuNAc linkage with expression of intermediate filaments. In studied normal adult epithelia, predominantly basal expression of alpha2,6NeuNAc and suprabasal expression of alpha2,3NeuNAc was seen, showing their differentiation-dependent linkage. The cells with markers of terminal differentiation (i.e. CK10+ alpha2,3NeuNAc+ alpha2,6NeuNAc-) prevailed in HNSCC, with increasing proportion from differentiation grade G1 to G3. High proportions of cytokeratin pCK37+ carcinoma cells occurred in all studied tumors and were accompanied by a hypersialylated phenotype (i.e. alpha2,3 NeuNAc+ alpha2,6NeuNAc+) not generally seen in non-transformed epithelia. It is hypothesized that these cells could be the pool for tumor spreading in the organism. Monitoring HNSCC using multiparameter phenotype analysis can produce new data important for the understanding of the biological behavior of HNSCC, and useful for the treatment rationalization.

Topics: Biotinylation; Carcinoma, Squamous Cell; Cell Differentiation; Cell Nucleus; Epithelium; Head and Neck Neoplasms; Humans; Keratins; Larynx; Lymphatic Metastasis; Mouth Mucosa; N-Acetylneuraminic Acid; Phenotype; Plant Lectins; Tissue Distribution; Tongue

To better detect occult cervical metastases.. RNA from 153 cervical lymph nodes was analyzed for the presence of squamous cell carcinoma using quantitative cytokeratin (CK) 14 real-time reverse transcription polymerase chain reaction (RT-PCR). Detection of CK RNA in pathologically negative nodes was further analyzed by semi-step sectioning and CK immunohistochemistry. Subjects Thirteen consecutive patients with head and neck squamons cell carcinoma (HNSCC) presenting to the Department of Otolaryngology/Head and Neck Surgery of the University of North Carolina at Chapel Hill for neck dissection.. Cytokeratin detection was deemed nonspecific if expressed at fewer than 50 molecules of CK 14 RNA per nanogram total RNA. Of 35 HNSCCs, 33 expressed CK 14 RNA, and 15 lymph nodes with routine pathologically positive metastasis were also positive for CK 14 RNA. Four lymph nodes that were pathologically negative nodes were positive for CK 14 RT-PCR, with 2 containing metastases detected by semi-step sectioning.. Cytokeratin 14 RT-PCR is very sensitive for detecting micrometastasis in lymph nodes that are negative by routine pathological examination, with a relatively high false-positive rate. Quantitative CK 14 RT-PCR could be used to identify nodes negative for tumor by standard pathological analysis that should be examined by step sectioning and CK immunohistochemistry. Identification of micrometastases in patients with HNSCC will allow for appropriate and aggressive treatment of patients with metastatic disease.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; False Positive Reactions; Female; Head and Neck Neoplasms; Humans; Keratins; Lymph Node Excision; Lymphatic Metastasis; Molecular Diagnostic Techniques; Neck Dissection; Neoplasm Staging; North Carolina; Prospective Studies; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm; Sensitivity and Specificity; Uterine Cervical Neoplasms; Women's Health

Our study was designed to find out the rate and the characteristics of micrometastasis in cervical lymph nodes using immunohistochemical staining.. From 69 patients, 1710 lymph nodes negative for metastasis on hematoxylin-eosin stain, were examined. Immunohistochemical stain was performed using pan-cytokeratin AE1/AE3 antibody.. In 13 cases, occult lymph node metastasis was detected by immunohistochemical method. On retrospective review of the hematoxylin-eosin stain by the pathologist, lymph node metastasis was detected in 4 of 13 patients.. Because the immunohistochemical method enhanced the detection rate of occult micrometastasis in cervical lymph nodes of head and neck squamous cell carcinoma patients, it may be recommended for routine diagnostic use in patient with negative for a lymph node metastasis on routine hematoxylin-eosin stain.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lymph Node Excision; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Retrospective Studies

We investigated the value of some tumor markers in the diagnosis and treatment follow-up of squamous cell carcinoma of the head and neck.. Ferritin, lipid-associated sialic acid (LSA), carcinoembryonic antigen (CEA), squamous cell carcinoma (SCC) antigen and CYFRA 21-1 levels were determined in 28 patients with squamous cell carcinoma and in a control group of 20 patients with benign lesions of the head and neck. The measurements were made before treatment and in the first and third months of treatment.. The sensitivity rates were as follows: 10.7% for ferritin, 89.3% for LSA, 21.4% for CEA, 42.9% for SCC antigen, and 14.3% for CYFRA 21-1. The specificity was 100% for all the markers. The sensitivity increased to 96.4% when CEA and LSA were used in combination. Declines in the levels after treatment were significant for ferritin, CEA, SCC antigen, and LSA. No significant relationship was found between the marker levels and lymph node metastasis, stage, and histologic differentiation of the tumors. Only ferritin and LSA levels were correlated with tumor size. Squamous cell carcinoma antigen was the only marker that manifested high levels in patients who developed locoregional recurrence and/or mortality.. Of the markers studied, LSA, CEA and SCC antigen may be of value in the evaluation of squamous cell carcinoma of the head and neck. Sensitivity and specificity rates may increase when they are used in combination.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Case-Control Studies; Female; Ferritins; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Lipids; Male; Middle Aged; N-Acetylneuraminic Acid; Predictive Value of Tests; Sensitivity and Specificity; Serpins

This paper attempts to evaluate the clinical usefulness of CYFRA 21-1 as a serum tumour marker in patients with head and neck squamous cell carcinoma (HNSCC). The serum concentration of CYFRA 21-1 was measured utilizing a new electrochemiluminescent immunoassay (ECLIA) in 142 patients with HNSCC before and after treatment, 68 patients with benign tumours of the head and neck, and 50 healthy controls. Serum levels of CYFRA 21-1 in patients with HNSCC were significantly higher than those of benign tumours and healthy controls (p < 0.001). The diagnostic sensitivity and specificity of CYFRA 21-1 for HNSCC were 62 per cent and 100 per cent, respectively. The positive rates of CYFRA 21-1 increased with progression of HNSCC, serum CYFRA 21-1 levels were related to the tumour stage expressed by primary tumour (T) and nodal status (N) (p < 0.001), but not related to patient age, gender, smoking and drinking habit, or histopathological grade (p > 0.05). Post-treatment levels of CYFRA 21-1 in HNSCC decreased significantly (p < 0.001). Among 38 patients with clinical or radiological evidence of a recurrence during follow-up, 78.9 per cent (30 of 38) showed an increase in CYFRA 21-1. The analytical ECLIA performance for serum CYFRA 21-1 provides a new means of clinical assessment for HNSCC. The results of ECLIA suggest that the serum marker CYFRA 21-1 is valuable not only for diagnosis but also for close monitoring of patients with HNSCC.

Topics: Adult; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunoassay; Keratin-19; Keratins; Luminescent Measurements; Male; Middle Aged

The frequency of amyloid deposits associated with squamous cell carcinoma (SCC) and dysplasia in the oral cavity, pharynx and larynx was examined. In addition, the origin of amyloid proteins by immunohistochemical staining with a panel of anticytokeratin monoclonal antibodies was investigated. Amyloid deposits were found in eight of 73 (11.0%) SCC and one of seven (14.3%) dysplasias in the oral cavity, in eight of 22 (36.4%) SCC and zero of two (0%) dysplasias in the pharynx, and in 22 of 37 (59.5%) SCC and four of 10 (40.0%) dysplasias in the larynx. Eight of 12 different cytokeratin (CK) antibodies reacted with these deposits: 34 beta E12 (CK1, -5, -10, -14) reacted with amyloid deposits in 19 of 19 cases (100%), LL002 (CK14) in eight of 18 cases (44.4%), MNF116 (CK5, -6, -8, -17) in eight of 19 cases (42.1%), D5/16B4 (CK5, -6) in five of 18 cases (27.8%), DE-K10 (CK10) in four of 17 cases (23.5%), RCK108 (CK19) in three of 18 cases (16.7%), 34 beta B4 (CK1) in three of 19 cases (15.8%) and AE8 (CK13) in two of 17 cases (11.8%). These antibodies always reacted with the cytoplasm of squamous cell lesions. Amyloid deposits in two cases contained a CK5 and CK14 pair, and in another two cases they contained both a CK5 and CK14 pair, and a CK1 and CK10 pair. Anti-CK antibodies, including OV-TL12/30 (CK7), c-51 (CK8), DC10 (CK18) and IT-Ks20.8 (CK20) did not react with the amyloid deposits. We conclude that the amyloid deposits associated with SCC or dysplasia in the oral cavity, pharynx or larynx were derived from CK of cancer cells and that some amyloid deposits might be assembled by two or more different CK.

Topics: Adult; Aged; Aged, 80 and over; Amyloid; Amyloidosis; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Pharyngeal Neoplasms; Precancerous Conditions

The incidence of occult nodal metastases associated with head and neck squamous cell carcinoma (HNSCC) and the clinical significance of nodal micrometastases by cytokeratin immunohistochemical analysis are examined.. In all, 1012 lymph nodes from 50 patients treated between 1992 and 2001 at the University of Colorado Health Sciences Center (Denver, CO) were evaluated retrospectively for micrometastases.. Serial sectioning in 5-to 6-microm interval specimens stained either with hematoxylin and eosin (H&E) or immunostaining for cytokeratins using the monoclonal antibody cocktail AE1/AE3 was performed in 21 N0, 11 N1, and 14 N2 patient cases. Cases that showed scattered cells with suspect staining qualities but without morphological features consistent with HNSCC were further evaluated by epithelial membrane antigen (EMA) immunohistochemical analysis.. H&E-stained and cytokeratin-stained sections revealed occult nodal micrometastases in 3.8% of N0 and 5% of N1 cases. Overall, 26 micrometastases were identified in N0 and N1 patients, causing 29% of N0 patients and 45% of N1 patients to be upstaged. Cytokeratin immunostaining detected micrometastases in eight cases that were negative on H&E serial sectioning. Serial sectioning by H&E alone identified three additional micrometastases. Negative EMA immunostaining confirmed the absence of malignant cells in lymph node sections that were equivocal on cytokeratin staining.. The use of serial sectioning with H&E and cytokeratin immunohistochemical analysis increases the detection of micrometastases that are often elusive by routine processing in patients with HNSCC. Improved methods of detecting micrometastases may provide a basis for improved planning of postoperative therapy for patients already at risk for tumor recurrence.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Culture Techniques; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Mucin-1; Neoplasm Metastasis; Neoplasm Staging; Spinal Neoplasms

Squamous cell carcinoma (SCC) is the most frequent malignant tumor of the upper aerodigestive tract. Cell lines of these tumors facilitate the investigation of various tumor biological parameters. This study was conducted to compare molecular biologic characteristics between cell lines and fresh tumor tissue.. In seven SCC-derived cell lines, cytokeratin 5/6 and cytokeratin 19 expression, DNA content, chromosome aberrations and tumorigenicity were assessed in nude rats. Unbalanced numerical and structural chromosomal aberrations were investigated by comparative genomic hybridization (CGH), and results were compared to those obtained in fresh tumor tissues of the same patients.. All cell lines expressed cytokeratins 5/6 and 19, indicating their epidermoid origin. Tumor growth after transplantation into nude rats occurred in five of seven cell lines. Routine histology and immunohistochemical examinations confirmed SCC. Aneuploidy was detected in all cell lines, with a 2c deviation index ranging from 1.9 through 9.5 and a 5c exceeding rate ranging from 2.6 through 36.7%. The most frequent chromosomal aberrations in cell lines were overrepresentations of chromosomal material on chromosomes 15q, 7p (5 cases each), 3q, 5p (4 cases each), and 11q and 17q (3 cases each) and losses of chromosomal material on chromosomes 3p, 18q (3 cases each), and 19p and 7q (2 cases each). Comparing these results to CGH analysis of fresh tumor tissue from the same patients, overrepresentations of chromosomal material on 10q, 20q and 21q, along with loss of chromosomal material on 4q was detected more frequently in primary tumors, whereas overrepresentations on 7p and loss of chromosomal material on 7q were more frequently detected in cell lines. Nevertheless, there was a high degree of similarity of chromosomal alterations in cell lines and corresponding fresh tumor tissue.. The data suggest a high degree of genetic similarity between tumor cells of cell lines and the tumors from which they were derived. Therefore, these cell lines can serve as an accurate model to investigate cell biology of SCC in vitro.

Topics: Animals; Biopsy; Carcinoma, Squamous Cell; Chromosome Aberrations; Chromosome Mapping; DNA, Neoplasm; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Nucleic Acid Hybridization; Papillomaviridae; Polymerase Chain Reaction; Rats; Rats, Nude; Transplantation, Heterologous; Tumor Cells, Cultured

This study was designed to help establish the most appropriate samples and tests to detect disseminated tumor cells (DTCs) for head and neck cancer patients.. Samples of bone marrow (BM) and central venous blood (CVB), collected preoperatively, and BM and peripheral venous blood, collected 3 months transcription postoperatively, were screened for the presence of DTCs using immunocytochemistry (ICC) with a pan-cytokeratin antibody and E48 reverse transcriptase-PCR. The molecular approach was also applied to intraoperative CVB.. The concordance between the molecular and ICC tests applied to preoperative samples, measured by Cohen's kappa, was not uniformly good, which likely reflected sampling errors, heterogeneity of E48 antigen expression, or stochastic effects. However, testing samples of BM and CVB preoperatively with the molecular or ICC approaches gave results that predicted disease-free survival and distant-metastases-free survival. Application of a single preoperative test predicted development of distant metastases, and the prediction could be improved by combining information derived from testing both CVB and BM. Applying two tests to the same sample of blood or BM served to validate the findings from a single assay but did not improve the prediction. Testing the intraoperative sample of CVB was also a sensitive predictor of distant metastases, but testing BM or blood 3 months postsurgery was not useful.. These findings suggest that detection of DTCs pre- or intraoperatively indicates a high risk of local and distant recurrence and reduced survival in head and neck cancer.

Topics: Bone Marrow; Bone Marrow Cells; Carcinoma, Squamous Cell; Cell Adhesion Molecules; Glycoproteins; GPI-Linked Proteins; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Neoplastic Cells, Circulating; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; Survival Rate; Treatment Failure; Tumor Cells, Cultured; Veins

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunoassay; Keratin-19; Keratins; Luminescent Measurements

Finding tumor markers for disease progression, and especially development of distant metastases, is desirable for patients with squamous cell carcinoma of the head and neck (SCCHN). Elevated serum levels of Cyfra 21-1 (cytokeratin fraction 21-1) have been frequently associated with disease progression in patients with lung cancer. In SCCHN, Cyfra 21-1 has not been established as a routine tumor marker yet, probably due to difficulties in finding the appropriate cut-off for the serum level. The aim of this study was to investigate whether assessment of changes in serum Cyfra 21-1 over time can predict distant metastases in patients with SCCHN, without attempting to establish an arbitrary cut-off for abnormal levels.. Cyfra 21-1 serum levels of 25 patients with SCCHN and distant metastases were evaluated by means of an ELISA test kit.. There was a wide range of Cyfra 21-1 serum levels at the time of primary diagnosis, without correlation with tumor size, lymph node status, time to recurrence, or presence of distant metastases. All patients had a clear increase of Cyfra 21-1 levels which preceded the appearance of distant metastases clinically.. Due to the wide range of Cyfra 21-1 levels at the time of primary tumor diagnosis, Cyfra-21-1 is neither a suitable screening marker for SCCHN, nor for diagnosis of distant metastases at the time of initial diagnosis of the tumor, but is of evident prognostic value for follow-up, especially for early detection of distant metastases.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease Progression; Enzyme-Linked Immunosorbent Assay; Follow-Up Studies; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Lymph Nodes; Neoplasm Metastasis; Neoplasm Recurrence, Local; Neoplasm Staging; Pharyngeal Neoplasms; Prognosis

A case of ectopic hamartomatous thymoma (EHT) arising in the supraclavicular region of a 52-year-old male is presented. The well-defined tumor measuring 1.7x1.5x0.7 cm consisted of three components: spindle cell (70%), epithelial (25%), and adipose (5%). The spindle cell component was characterized by sheet-like, haphazard and short fascicular arrangements of bland spindle cells. Neither mitotic figures nor cellular pleomorphism were found. Admixed with, and adjacent to, the spindle cell areas was an obviously epithelial component of variable appearance, ranging from glandular spaces lined by mainly cuboidal clear cells, irregularly anastomosing cords, and strands of epithelial cells to irregular solid nests of squamous epithelium with dark and clear cytoplasm. Myoepithelial cells were also observed. Immunohistochemically, the spindle cells were strongly and diffusely positive for cytokeratins and some of them were positive for BRST2, alpha-smooth muscle actin, and CD10. The tumor was negative for S-100 protein, glial fibrillary acidic protein, and CD34. Ultrastructurally, tonofilaments and desmosomes were observed in the spindle cells. The findings indicate an epithelial origin. The patient was well without recurrence or metastasis 8 months after excision. Pathologists and clinicians should be aware of the existence of ectopic hamartomatous thymoma in the supraclavicular or suprasternal region and should differentiate it from a high-grade sarcoma, such as biphasic synovial sarcoma or glandular malignant peripheral nerve sheath tumor.

Topics: Actins; Apolipoproteins; Apolipoproteins D; Carrier Proteins; Glycoproteins; Hamartoma; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Membrane Transport Proteins; Microscopy, Electron; Middle Aged; Neprilysin; Thymoma

To apply a new immunocytochemistry (ICC) assay to peripheral blood samples for micrometastatic circulating tumor cell detection in patients with head and neck squamous cell cancer (HNSCC).. The ICC assay uses established monoclonal antibodies that bind to tumor-associated antigens combined with an enrichment system that uses positive selection with anti-human epithelial antigen (EpCAM antibody) to detect circulating tumor cells.. Eighteen consecutive patients newly diagnosed as having HNSCC are described.. Of the 18 patients, 8 (44%) demonstrated circulating tumor cells using the ICC assay. The numbers of patients positive for circulating tumor cells per stage are as follows: stage I, 1 of 1; stage II, 0 of 2; stage III, 2 of 5; stage IV, 5 of 6; and unknown stage, 0 of 4. The numbers of patients positive for circulating tumor cells per location are as follows: oral cavity, 1 of 2; oropharynx, 3 of 4; glottic area, 3 of 5; supraglottic area, 1 of 3; and unknown primary 0 of 4.. Circulating tumor cells were identified in almost half of the patients using the ICC assay. In a literature review, we were not able to identify previous reports of circulating tumor cell detection in patients with HNSCC from peripheral blood samples using ICC or identify any study that has attempted to quantify circulating tumor cell levels. Although the clinical implications of circulating tumor cells in micrometastatic tumor detection in patients with HNSCC are still unknown, they may be significant. Long-term follow-up may help elucidate the patients in whom conventional treatment may fail and, thus, those who may benefit from different treatment; it may also assist with the detection of recurrence with a simple blood collection.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplastic Cells, Circulating

It has been previously shown that S100A2 is downregulated in tumor cells. The level of immunohistochemical S100A2 expression was therefore characterized in 424 normal and tumoral (benign and malignant) tissues of various origins, but mostly epithelial (with either glandular, squamous, respiratory or urothelial differentiation). We also investigated whether S100A2 could be co-localized with cytokeratin K14, an intermediate filament protein expressed in basal proliferative keratinocytes. Our data show that S100A2 has a low level of expression in non-epithelial tissue. In epithelial tissue S100A2 expression decreases remarkably in the tumors when compared to the normal specimens, and was correlated with the level of keratin K14. This decrease in S100A2 staining from normal to cancer cases is more pronounced in glandular than in squamous epithelial tissue. In addition, the patterns of S100A2 staining also differ between glandular and squamous tissue. These data suggest distinct functional roles for S100A2 in epithelial tissue of squamous or glandular origins.

Topics: Carcinoma, Squamous Cell; Chemotactic Factors; Cytoplasm; Epithelial Cells; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Neoplasms, Glandular and Epithelial; Paraffin Embedding; S100 Proteins; Tissue Fixation

Cyfra 21-1, measuring serum fragments of cytokeratin 19, has been found to be related to tumour stage and tumour size in patients with cervical cancer. It could be a promising marker in squamous lung cancer. We evaluated this new marker with carcinoembryonic antigen, (CEA) and squamous cell carcinoma antigen (SCC-Ag) in the monitoring of 27 patients with head and neck cancer.. The retrospective study group consisted of 27 patients, 17 not suited for surgery and 10 after laser resection. Patients were clinically staged according to the TNM-classification. The mean age of the patients was 53 years (range 37-70 years). Serum levels of each marker were studied in relation to tumour stage and clinical status of the patients during radiotherapy and 6 weeks after the end of the treatment. The clinical performance of the various assays to separate those patients with complete remission from those patients with the presence of tumour was assessed.. Pre-treatment serum Cyfra 21-1, CEA, and SCC-Ag levels were not related to stage of disease and were not found to be predictive of tumour response. The clinical performance of post-treatment serum SCC-Ag levels in predicting the presence of tumour was not better than the Cyfra 21-1 assays.. We could not conclude from this study that Cyfra 21-1 marker is an additional parameter in identifying patients at risk of residual tumour after treatment, recurrent or progressive disease. An elevation of cyfra 21-1 marker was not detectable in 70% of the cases with macroscopic tumour. Therefore, Cyfra 21-1 is not a reliable parameter for the monitoring of patients with head and neck cancer during radiotherapy.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Follow-Up Studies; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratin-19; Keratins; Middle Aged; Radiotherapy Dosage; Retrospective Studies; Risk Factors; Serpins; Time Factors

Topics: Adult; Diagnosis, Differential; Epidermal Cyst; Head and Neck Neoplasms; Humans; Keratins; Magnetic Resonance Imaging; Male; Skin Diseases

The definition of biological markers for oropharynx and larynx cancer is essential to predict their clinical behavior. Since cellular glycans play an important role in biological information transfer, we have employed an endogenous lectin, galectin-3, to examine in primary squamous carcinomas, lymph node metastases, and physiological squamous epithelia whether glycans recognized by this lectin are altered in relation to the state of differentiation. The expression of galectin-3 was concomitantly evaluated by immunohistochemistry using the A1D6 monoclonal antibody. In addition, other antibodies were used for the detection of cytokeratins and desmosomal proteins (desmoplakin-1 and desmoglein). The results show the expression of galectin-3-reactive ligands in moderately/highly differentiated carcinomas only in areas exhibiting a high level of keratinization. Except for one patient out of 14, metastatic cells in lymph nodes expressed no accessible binding sites for galectin-3. No galectin-3-reactivity was detected in the basal cell layer of all studied normal epithelia (which contains the proliferating cells). The suprabasal layers were positive in epidermis and epithelium of tongue and cornea and negative in epithelium of palatine tonsil. The tumor cells expressed galectin-3 with an intensity positively correlated with tumor differentiation. The position of galectin-3-reactive sites colocalized with the two tested desmosomal proteins. However, presence of these proteins was also detected in areas of tumor and suprabasal layers of tonsil epithelium where no binding reactivity for galectin-3 was found. The present study showed that expression of galectin-3-reactive glycoligands is differentiation-dependent in normal as well as malignant squamous cells. Colocalization of galectin-3-reactive sites with desmosomal proteins (desmoplakin-1 and desmoglein) suggests an association of the galectin-3 ligand(s) with the cell surface, pointing to a potential participation of galectin-3 in mediation of intercellular contacts in these tumor types.

Topics: Antigens, Differentiation; Binding Sites; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Cytoskeletal Proteins; Desmogleins; Desmoplakins; Epithelial Cells; Fluorescent Antibody Technique, Indirect; Fluorescent Dyes; Galectin 3; Head and Neck Neoplasms; Humans; Keratins; Ligands; Lymphatic Metastasis; Membrane Glycoproteins; Neoplasm Proteins; Neoplasm Staging; Prognosis

Glandular schwannoma is a rare variant of schwannoma characterized by the presence of glands in an otherwise typical schwannoma. We report a patient with benign glandular schwannoma occurring on the scalp, a site not previously reported. Histologically, a well-defined, encapsulated oval nodule was observed in the subcutaneous tissue. The nodule was composed of a spindle cell component and glandular structures. The spindle cell component stained positively for S-100 protein. All of the glandular epithelium stained with CAM 5.2 and epithelial membrane antigen but not with S-100 protein. The glandular epithelium was focally positive for carcinoembryonic antigen. The histogenesis of the glandular elements in these tumours is still debated. The variable size of the glandular structures in our case was evidence against an entrapped normal sweat gland origin. The glandular epithelium did not stain with S-100 protein at all, but stained with CAM 5.2, which did not support a direct metaplastic origin of the epithelial elements from the schwannian component. A few scattered CAM 5.2-positive cells and microglandular structures in our case may be the initial differentiating epithelial elements possibly derived from pluripotential neural crest cells.

Topics: Adult; Biomarkers; Biomarkers, Tumor; Female; Head and Neck Neoplasms; Humans; Keratins; Neoplasm Proteins; Neurilemmoma; Scalp; Skin Neoplasms

A 71-year-old woman had a dome-shaped, slightly erythematous nodule on the anterior scalp. The nodule histopathologically revealed sebaceoma based on the silhouette and cytology. A notable and unique finding was often observed in the aggregations of sebaceoma; an arrangement of small, monomorphous, cigar-shaped basaloid cells in linear rows parallel to one another, resembling the palisading of nuclei of Verocay bodies, namely a rippled-pattern. Although we are not certain that sebaceoma can be clearly separated from trichoblastoma with sebaceous differentiation in all cases, in the present case, the absence of an abundant and densely fibrotic stroma, of follicular differentiation, and of a palisading border in the neoplastic aggregations as well as the presence of many vacuolated cells and tiny duct-like spaces favors the diagnosis of sebaceoma rather than trichoblastoma with sebaceous differentiation. Based on the expression patterns of CKs as well as similar cytological features between germinative cells in our case and immature cells in the mantles of normal vellus follicles, we believe that rippled-pattern sebaceoma is composed of immature sebaceous germinative cells with some foci of advanced sebaceous differentiation (toward the sebaceous duct and sebaceous lobule).

Topics: Aged; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Scalp; Sebaceous Gland Neoplasms

Specimens from 17 head and neck tumor patients were immunohistochemically stained with monoclonal antibodies against HLA-DR, CD1a, RFD1, LAG, CD3, CD4, CD8, CD45RO, CD68, and cytokeratin to identify the nature and distribution of dendritic cells (DCs), T cells, and macrophages. Small numbers of DCs were present in all but 2 specimens. They were located between the tumor cells and in the stroma, especially in areas of inflammatory cell infiltration. Variable numbers of T lymphocytes (cytotoxic and memory type) occurred in the same locations. Numerous macrophages were found in the epithelium, in the stroma, and in the vicinity of tumor cells. The presence of DCs in head and neck tumors indicates that the organism has activated the immune surveillance system and is trying to present tumor antigens. Considering the sparsity of DCs in the malignant tissues, the T cell response can be only limited.

Topics: Antibodies, Monoclonal; Antigens, CD; Carcinoma; Dendritic Cells; Head and Neck Neoplasms; HLA-DR Antigens; Humans; Immunohistochemistry; Keratins; Langerhans Cells; Major Histocompatibility Complex

Remnants of the branchial apparatus can produce lesions in the head and neck region in later life, often amenable to fine-needle aspiration (FNA) diagnosis. Yet such remnants or rudimentary lesions can remain clinically undetected and can later interfere with the cytologic interpretation of other deep lesions of the neck, as the present case demonstrates. In this case the lesion, which by a subsequent resection turned out to be a neurilemmoma, had been adequately sampled by the FNA, yet the cytologic diagnosis was sidetracked by the presence in the specimen of immature squamous epithelial tissue fragments and other elements (multinucleated histiocytes, calcifications), on the basis of which the diagnosis of an epithelial lesion, likely malignant, was made. The neck surgery and a preceding endoscopic examination of the mouth, pharynx, and larynx did not identify such a lesion, but a detailed microscopic examination of the fibroadipose tissue between the tumor and the peripharyngeal region revealed the presence of epithelial microfragments with morphology partly corresponding to that of the FNA cytology, highly indicative of a branchiogenic lesion in the peripharyngeal region. The basic embryology of the branchial apparatus resulting in such defects is presented, as well as tentative guidelines for recognizing material deriving from accidental sampling of such lesions during FNA investigations of deep-seated masses of the neck. Diagn. Cytopathol. 2000;22:157-160.

Topics: Aged; Biopsy, Needle; Branchial Region; Diagnostic Errors; Epithelial Cells; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Neurilemmoma

To discover unique genes specific for squamous cell carcinoma of the head and neck for eventual development as tumor markers and vaccine candidates.. Molecular biological analysis of fresh-frozen head and neck squamous cell cancer (HNSCC).. A subtractive library was made from two HNSCC and six normal tissues using a polymerase chain reaction (PCR)-based approach. Genes from this library were PCR amplified and placed on a microarray glass slide. RNA was prepared or obtained from 16 fresh-frozen HNSCC and 22 normal tissue sources. Fluorescent probes were made from the polyA+ RNA derived from the tumor and normal tissues. The probes were hybridized to the glass slides and excited by a tuneable laser. One hundred seven of the genes showing the highest differential fluorescence value between tumor and normal tissue were identified by sequence analysis.. Thirteen independent genes were found to be overexpressed in tumor tissues. Of these, nine were previously known: keratins K6 and K16, laminin-5, plakophilin-1, matrix metalloproteinase-2 (MMP), vascular endothelial growth factor, connexin 26, 14-3-3 sigma, and CaN19. The level of polyA+ RNA of these genes in the tumors was significantly different from the levels in normal tissue (P < .05). Four previously unidentified genes were also discovered to have increased expression in tumor tissue. Comparing the total tumor group (n = 16) to the normal group (n = 22), only one of these genes showed significant overexpression.. We report the identification of nine known genes that are significantly overexpressed in HNSCC as compared to normal tissue using subtractive and microarray technology. In addition, we present four previously unidentified genes that are overexpressed in a subset of tumors. These genes will be developed as tumor markers and vaccine candidates.

Topics: 14-3-3 Proteins; Adolescent; Adult; Aged; Biomarkers, Tumor; Cancer Vaccines; Carcinoma, Squamous Cell; Cell Adhesion Molecules; Chemotactic Factors; Connexin 26; Connexins; DNA Probes; DNA, Complementary; Endothelial Growth Factors; Gene Expression Regulation, Neoplastic; Gene Library; Head and Neck Neoplasms; Humans; In Situ Hybridization, Fluorescence; Kalinin; Keratins; Lasers; Lymphokines; Matrix Metalloproteinase 2; Middle Aged; Molecular Biology; Plakophilins; Polymerase Chain Reaction; Proteins; RNA, Messenger; RNA, Neoplasm; S100 Proteins; Sequence Analysis, DNA; Tyrosine 3-Monooxygenase; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

The use and limitations of fine-needle aspiration (FNA) of lesions of the parotid gland are known, but those of nonparotid lesions of the head have been described only sporadically. We conducted this study to evaluate the utility of FNA and to analyze the causes of diagnostic discrepancies for these lesions. A total of 6,898 FNAs of different sites was performed at our institutions between January 1991-August 1998, and 214 (3.1%) of the cases were FNAs of nonparotid lesions of the head. The most common diagnosis of nonparotid lesions was squamous-cell carcinoma, in 22% (n = 48), and the most common site aspirated was the scalp, in 34% (n = 73). Lipomas and keratinous cysts comprised 5% (n = 9) of the total. A statistical analysis was conducted on 98 paired cytology and histology (n = 83) and cytology and flow cytometry (n = 15) specimens (70 malignant and 28 benign). FNA recognized the malignant and benign nature of the lesion in 60 and 26 cases, respectively with 86% sensitivity 93% specificity and 88% accuracy. Causes of false-negative FNA diagnoses (n = 10) included sampling error (n = 6), bloody smears with scant cellularity (n = 3), and bland cytomorphology (n = 1). Florid granulation tissue and a mucocele of the tongue accounted for the two false-positive cases. We conclude that FNA is an effective tool for triage of surgery candidates with nonparotid lesions of the head. Adequate samples with sufficient cellularity are required for avoiding false-negative diagnoses. Occasionally, tissue biopsy is needed for diagnosis of equivocal cases.

Topics: Adult; Aged; Aged, 80 and over; Biopsy, Needle; Carcinoma, Squamous Cell; Epidermal Cyst; Evaluation Studies as Topic; Female; Head and Neck Neoplasms; Humans; Keratins; Lipoma; Male; Middle Aged; Multiple Myeloma; Scalp; Sensitivity and Specificity

Oral ulcerative mucositis is a common toxicity associated with drug and radiation therapy for cancer. It impacts on quality of life and economic outcomes, as well as morbidity and mortality. Mucositis is often associated with dose limitations for chemotherapy or is a cause for dose interruption for radiation. The complexity of mucositis as a biological process has only been recently appreciated. It has been suggested that the condition represents a sequential interaction of oral mucosal cells and tissues, pro-inflammatory cytokines and local factors such as saliva and the oral microbiota. The recognition that the pathophysiology of mucositis is a multifactorial process was partially suggested by the observation that interleukin-11 (IL-11), a pleotropic cytokine, favorably altered the course of chemotherapy-induced mucositis in an animal model. In the current study, we evaluated a series of biologic and morphologic outcomes to determine their roles and sequence in the development of experimental radiation-induced mucositis and to evaluate the effects of IL-11 in attenuating them. Our results suggest that IL-11 favorably modulates acute radiation-induced mucositis by attenuating pro-inflammatory cytokine expression. Data are also presented which help define the pathobiological sequence of mucositis.

Topics: Animals; Apoptosis; Cricetinae; Disease Progression; Head and Neck Neoplasms; Immunohistochemistry; Interleukin-1; Interleukin-11; Keratins; Male; Mast Cells; Mesocricetus; Mouth Mucosa; Oral Ulcer; Radiation Injuries, Experimental; Stomatitis

This study examines a new tumour marker, Cyfra 21-1, as a prognostic marker in predicting the survival of H&N cancer patients, and its correlation with clinical outcome during prolonged follow up of these patients. The study included 67 patients with primary detection of carcinoma of H&N. The survival of these patients was evaluated in correlation with the disease stage and Cyfra 21-1 levels at initial diagnosis. 38 patients were followed clinically and with serial assays for at least 12 months, or until recurrence was diagnosed. Cyfra 21-1 levels were determined periodically, using an Elisa kit. Patients with Cyfra 21-1 < 1.5 ng ml(-1)had a higher survival rate compared to patients with Cyfra 21-1 > or = 1.5 ng ml(-1)(63% vs. 20%, respectively). The risk ratio of Ln(Cyfra 21-1) is 1.62 (P = 0.028). In a Cox regression model that included the disease stage and Ln(Cyfra 21-1), Ln(Cyfra 21-1) was preferred as the main parameter for predicting patients survival. In 83% of the 12 patients with recurrent or residual disease, Cyfra 21-1 was elevated before or during clinical detection of the recurrence. Cyfra 21-1 was found to be a prognostic marker for carcinoma of H&N, unrelated to the stage of the disease. Elevated levels of Cyfra 21-1 without clinical evidence of disease can be attributed to the marker's mean lead-time as compared to the clinical appearance of the disease.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Predictive Value of Tests; Prognosis; Survival Analysis; Survival Rate

Serum concentrations of Cyfra 21-1, a soluble cytokeratin fragment, were evaluated in 157 healthy patients, 66 patients with benign ear, nose or throat disease, and 102 patients with untreated epidermoid carcinoma of the head and neck. The technique had a sensitivity of 53.9% and specificity of 98.4% for a cutoff value of 1.5 ng/ml. Cyfra 21-1 levels differed significantly with tumor location (p < 0. 005) and tumor stage (p < 0.005), but not with the degree of histological differentiation.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Sensitivity and Specificity

The purpose of this study is to better estimate the true incidence of occult regional metastases associated with stage I and II squamous cell carcinoma of the oral cavity. The clinical and prognostic significance of micrometastatic disease discovered by cytokeratin immunoperoxidase reactivity in the previously pathologically N0 neck is also evaluated.. Forty patients treated between 1985 and 1996 with T1 or T2 squamous cell carcinoma of the lip and oral cavity were studied. All had primary surgical treatment including functional neck dissection. No metastases were demonstrated on hematoxylin and eosin microscopy. All specimens were reexamined with immunoperoxidase staining for cytokeratin.. Five percent of patients had micrometastatic disease. Retrospective analysis of patients with a minimum follow-up of 2 years has failed to show a statistically significant association between a positive cytokeratin analysis and poor locoregional control or overall survival.. Results suggest that the true incidence of occult metastases with carcinoma of the oral cavity is significantly higher than previously documented. However, the prognostic significance of these findings remains unclear.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Incidence; Keratins; Male; Middle Aged; Mouth Neoplasms; Neck Dissection; Neoplasm Staging; Prognosis; Retrospective Studies; Survival Analysis

The preventive effects of retinoids on oral carcinogenesis may be related to their ability to modulate the growth and differentiation of human oral squamous epithelial cells. Nuclear retinoid receptors (RAR alpha, beta, and gamma, and RXR alpha, beta, and gamma) may mediate these effects by regulating gene transcription. The removal of serum from the growth medium of two head and neck squamous cell carcinoma lines 1483 and SqCC/Y1 resulted in a decrease in RAR beta mRNA level and concurrent increases in the expression of the keratin K1 and transglutaminase type I (TGase I), which are markers of differentiation of keratinizing squamous epithelial cells. All-trans-retinoic acid (tRA) or 13-cis-RA increased RAR beta and decreased K1 and TGase I mRNA levels in serum-free medium. Transcriptional activation of reporter genes by means of retinoid response elements (RARE and RXRE) indicated that the RXR-RAR pathway predominates over the RXR homodimer pathway in the 1483 cells. Among several synthetic retinoids with preference for binding to specific nuclear retinoid receptors, those that induced RAR beta also suppressed K1. The inverse association between RAR beta expression and K1 and TGase I levels implicates this receptor in suppression of keratinization in oral epithelial cells.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Differentiation; Cell Division; Collagenases; Dose-Response Relationship, Drug; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Keratins; Promoter Regions, Genetic; Receptors, Retinoic Acid; Response Elements; Retinoid X Receptors; Retinoids; Transcription Factor AP-1; Transcription Factors; Transcription, Genetic; Transcriptional Activation; Tretinoin; Tumor Cells, Cultured

The development of aneuploid clones from diploid progenitor cells is a regular characteristic of head and neck squamous cell carcinoma progression. While the significance of aneuploidy formation for the acquisition of invasive and metastatic behavior is well documented, little is known about the contribution of diploid tumor cells after aneuploid clones have emerged. To distinguish diploid cells of epithelial origin from benign cellular components, we applied multiparameter flow cytometry of DNA content and cytokeratin (CK) expression to 36 primary tumors. Twenty-seven carcinomas accommodated aneuploid cell lines that stained positive for CK. All diploid cell populations obtained from aneuploid carcinomas contained CK-positive subpopulations as did all of nine tumors that consisted exclusively of diploid cells. The proportions of CK-positive diploid cells ranged between 6% and 80%, independent of whether they were achieved from entirely diploid or from aneuploid carcinomas. CK-gated diploid and aneuploid cell populations showed largely identical S-phase fractions. These results emphasize that diploid tumor cells regularly persist after the development of aneuploid clones and significantly contribute to local tumor progression. Despite the presence of diploid epithelial cells in aneuploid primary tumors, exclusively the aneuploid clones of eight corresponding lymph node metastases were CK-positive. This provides further evidence of a largely reduced metastatic potential of diploid tumor cells.

Topics: Aneuploidy; Carcinoma, Squamous Cell; Clone Cells; Diploidy; Disease Progression; Epithelial Cells; Flow Cytometry; Head and Neck Neoplasms; Humans; In Vitro Techniques; Keratins

DNA ploidy and cell-cycle distribution were determined by flow cytometry in fresh tumor tissue of 27 epithelial head and neck carcinomas. Epithelial cells were labeled with a fluorescein-isothiocyanate-conjugated cytokeratin antibody to study the possible influence of contaminating stromal and inflammatory cells on the results of cell-cycle analysis of tumor cells. The patient sample included 26 men and 1 women with a mean age of 60 years. Without cytokeratin gating, 11/27 tumors (40.74%) were aneuploid. After selecting the cytokeratin population, 10 more aneuploid tumors were found that had not been detected when considering the total population. Therefore, aneuploid tumors increased from 40.74% to 77.74%. The remaining 6/27 (22.26%) tumors were diploid. In the tumors that were either aneuploid without cytokeratin gating or diploid, the S-phase and G2M phase were significantly higher after cytokeratin staining, specially in diploid tumors (24.2% versus 10% and 6.8% versus 3.2%, respectively, p < 0.01). Therefore, in head and neck tumors cytokeratin staining optimizes both the determination of DNA ploidy and cell-cycle analysis, which is advantageous for tumor staging and prognosis assessment in these patients.

Topics: Adult; Aged; Antibodies, Monoclonal; Antibodies, Neoplasm; Carcinoma, Squamous Cell; Diploidy; DNA, Neoplasm; Female; Flow Cytometry; Head and Neck Neoplasms; Humans; Interphase; Keratinocytes; Keratins; Male; Middle Aged; Quantitative Trait, Heritable

The serum concentrations of three separate tumor markers, squamous cell carcinoma antigen (SCC Ag), carcinoembryonic antigen (CEA) and Cyfra 21-1 were clinically correlated in 86 randomly selected patients with squamous cell carcinoma involving the head and neck. Positive findings for each tumor marker were totalled and statistically analysed. The upper limits of normal for SCC Ag, CEA and Cyfra 21-1 were set at 1.5, 2.5 and 2.0 ng/ml, respectively. Positivity rates were 20.6% for SCC Ag, 14.0% for CEA and 41.7% for Cyfra 21-1. Elevated Cyfra 21-1 concentrations correlated somewhat with age, whereas elevated CEA levels correlated with the site of tumor involvement. Overall, Cyfra 21-1 appeared to be the most useful marker in head and neck squamous cell carcinoma.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratin-19; Keratins; Male; Middle Aged; Radioimmunoassay; Random Allocation; Serpins

This study is designed to assess gene expression of squamous cell carcinoma antigen (SCCA) mRNA to detect micrometastases in cervical lymph nodes (LNs) of head and neck cancer. We examined the expression of SCCA mRNA in 12 primary tumors and 212 cervical LNs (101 LNs taken from 8 patients with tongue cancer, 71 from 7 patients with gingival cancer, 19 from 2 patients with laryngeal cancer, 9 from 2 patients with pharyngeal cancer, 7 from 1 patient with cancer of the buccal mucosa, and 5 from 1 patient with cancer of floor of the mouth). Detectability of metastatic LNs by nested and single reverse transcriptase-polymerase chain reaction (RT-PCR) was compared with semiserial sections (hematoxylin-eosin staining and keratin immunostaining). All primary tumors expressed SCCA mRNA. Of 198 histologically metastasis-negative nodes, SCCA mRNA was detected in 37 (18.7%) by nested PCR. Eleven micrometastatic foci in 9 LNs (4.6%) were discovered by semiserial sectioning. This suggests that SCCA mRNA is a promising tumor marker for detecting the micrometastases in cervical LNs of head and neck cancer.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Biopsy; Carcinoma, Squamous Cell; False Negative Reactions; Gingival Neoplasms; Head and Neck Neoplasms; Humans; Keratins; Laryngeal Neoplasms; Lymphatic Metastasis; Microtomy; Mouth Mucosa; Neck; Neoplasm Proteins; Pharyngeal Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Sensitivity and Specificity; Serpins; Staining and Labeling; Tongue Neoplasms

We report the development of a reverse-transcriptase polymerase chain reaction assay that detects (in paraffin-embedded, formalin-fixed tissue) the SYT-SSXchimeric RNA transcript resulting from the t(X;18) of synovial sarcoma. The primers chosen detect both of the SSX1 and SSX2 partners, and the target sequence is small enough (87 base pairs) to be reliably detected in archival and variably processed consultation material. To demonstrate its usefulness, we applied it to 14 problematic cases, including spindle cell tumors of the thoracic region, of the neck, and of subcutaneous tissue. For instance, we show that, depending on the location, synovial sarcoma can mimic malignant solitary fibrous tumor, the spindle epithelial tumor with thymus-like differentiation, or skin adnexal tumors. Molecular detection of the SYT-SSX chimeric RNA should allow the reclassification of difficult cases in which the morphologic features overlap different entities or in which tumor nosology is still evolving.

Topics: Adult; Aged; Chromosomes, Human, Pair 18; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Molecular Probe Techniques; Neoplasm Proteins; Paraffin Embedding; Polymerase Chain Reaction; Proteins; Proto-Oncogene Proteins; Recombinant Fusion Proteins; Repressor Proteins; RNA, Neoplasm; Sarcoma, Synovial; Skin Neoplasms; Thoracic Neoplasms; Transcription, Genetic; Translocation, Genetic; X Chromosome

CYFRA 21-1 (CYFRA) is a newly developed tumour marker which is useful in evaluating large cell lung carcinoma, especially the squamous cell type. The purpose of this study was to assess the clinical value of CYFRA for squamous cell carcinoma of the head and neck and compare the results with squamous cell carcinoma antigen (SCCA). Serum levels of CYFRA were measured in 168 patients with a newly diagnosed head and neck squamous carcinoma. In addition, 77 patients without evidence of neoplasm were included as controls. At the same time, SCCA was also determined. The cut-off values of CYFRA and SCCA, determined at the 95th percentile of the standard Gaussian variate of controls, were 2.48 ng/ml and 1.49 ng/ml respectively. The diagnostic sensitivity of CYFRA was superior to that of SCCA, especially for nasopharyngeal carcinoma. The sensitivity of CYFRA for nasopharyngeal carcinoma was much higher (58.3%) than that of SCCA (15.5%). However, the sensitivity of CYFRA is not satisfactory in all types of squamous carcinoma. For oral cancer, the sensitivity is only 25.6%. CYFRA is a useful serum marker for patients with certain types of head and neck squamous carcinoma, such as nasopharyngeal carcinoma and hypopharyngeal carcinoma. In addition, CYFRA may be also useful in monitoring recurrence of certain types of SCCHN, which are sometimes difficult to detect.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Case-Control Studies; Female; Head and Neck Neoplasms; Humans; Immunoradiometric Assay; Keratin-19; Keratins; Male; Middle Aged; Sensitivity and Specificity; Serpins

Metastatic spread to cervical lymph nodes is a major determinant of outcome in head and neck cancer. One hundred and ninety-six lymph nodes dissected from fresh surgical specimens from 24 patients with primary head and neck squamous cell carcinoma (SCC) were bisected. Messenger RNA (mRNA) extracted from one half and from a segment of the primary tumour was amplified by reverse transcriptase (RT)-polymerase chain reaction (PCR) with primers flanking the fifth intron of human type II keratin K5. DNA bands resolved by agarose gel electrophoresis were confirmed as specific transcripts by sequencing. The other half of each node was fixed in formalin for histology and, in selected nodes, for immunohistology for cytokeratins. Of 153 nodes suitable for analysis, 14 nodes contained metastatic tumour detected by light microscopy and also tested positive for K5 mRNA by RT-PCR. Fifty-six nodes were histologically negative for tumour but positive for K5 mRNA, and 83 nodes were negative for both histology and K5 mRNA. Extracts of the primary tumour always reacted positively for K5 by RT-PCR, whereas lymph nodes from patients without malignancies, and blood lymphocytes from a healthy volunteer reacted negatively. RT-PCR designed to detect the presence of processed transcripts of type II keratin K5 in stratified squamous epithelial cells may be a sensitive technique to detect the presence of viable and potentially metastatic carcinoma cells in lymph nodes draining head and neck SCC.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lymphatic Metastasis; Neck; Polymerase Chain Reaction; RNA Splicing; RNA, Messenger; RNA, Neoplasm; Sensitivity and Specificity; Sequence Analysis, RNA

To investigate the utility of primer extension preamplification (PEP) in the genetic analysis of head and neck squamous tumorigenesis, microsatellite analysis was performed on matched deoxyribonucleic acid (DNA) samples extracted from 32 flow-sorted and microdissected specimens before and after PEP. Eighteen fresh and nine archival specimens were taken from invasive carcinomas, and five specimens were obtained from microdissected archival premalignant squamous epithelial lesions. Identical microsatellite patterns were observed in 276 (87%) of the 319 paired PEP and non-PEP genotypes with sufficient DNA. Overall, 13 (4%) of the PEP and 28 (8.8%) of the non-PEP fresh tissue samples failed specific microsatellite amplification. All 14 PEP-archival specimens were successfully amplified. Sorted cells showed a higher incidence (42.8%) of loss of heterozygosity (LOH) in both PEP and non-PEP samples compared with their unsorted counterparts. The results of this study indicate that (a) PEP is a simple and reliable technique for enhancing the DNA yield from small specimens; (b) flow sorting, in certain cases, improves the interpretation of genetic results; and (c) PEP may be used to compensate for PCR failure of unamplified DNA specimens in these lesions.

Topics: Carcinoma, Squamous Cell; DNA Mutational Analysis; DNA Primers; DNA, Neoplasm; Flow Cytometry; Genome, Human; Head and Neck Neoplasms; Humans; Keratins; Loss of Heterozygosity; Microsatellite Repeats; Molecular Probe Techniques; Neck Dissection; Polymerase Chain Reaction; Respiratory Tract Neoplasms

Topics: Biomarkers, Tumor; Cytodiagnosis; Head and Neck Neoplasms; Humans; Keratins; Neoplasms, Second Primary

We report an 85-year-old man with squamous cell carcinoma on the right pinna. Two years after the excision of the lesion, metastatic foci were found extending from the right retromandibular to the mastoid region and into the parapharyngeal space. Histopathologically, the primary tumor showed interconnecting nests of atypical cells invading into the dermis from multiple epidermal and infundibular foci. The tumor had both squamous and glandular differentiation. A peculiar finding was the presence of decapitation secretion in the glandular foci. To our knowledge, definite apocrine differentiation in squamous cell carcinoma has not previously been reported.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Apocrine Glands; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Cell Differentiation; Ear Neoplasms; Ear, External; Head and Neck Neoplasms; Humans; Keratins; Male; Mastoid; Neoplasm Invasiveness; Pharyngeal Neoplasms; Skin Neoplasms; Skull Neoplasms

Soluble intercellular adhesion molecule-1 (s-ICAM-1) was measured in the sera of 131 patients with primary and 50 patients with recurrent squamous cell cancer of the head and neck (HNSCC). 30 patients with benign ear, nose and throat diseases served as controls. s-ICAM-1 levels in serum are high in patients with HNSCC, particularly in the advanced tumor stages (UICC IV). Highest levels can be measured at the time of tumor recurrence and locoregional lymph node metastases. The sensitivity (95% specificity) of s-ICAM-1 (cutoff-level: 473 ng/ml) is 4% at primary diagnosis and 12% for recurrent disease. A coefficient of correlation for s-ICAM-1 in combination with SCC, carcinoembryonic antigen and CYFRA 21-1 indicates that no correlation can be found of s-ICAM-1 compared with traditional tumor markers. Due to overlapping values in control and patient groups s-ICAM-1 is not suitable for a specific clinical use in HNSCC.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Intercellular Adhesion Molecule-1; Keratins; Lymphatic Metastasis; Prognosis; Serpins

A case of heterotopic salivary gland adenocarcinoma (HSGA) in the right cervical region is presented. The carcinoma cells were positive for alpha-amylase, carcinoembryonic antigen, epithelial membrane antigen, cytokeratin as well as for expression of human salivary alpha-amylase messenger ribonucleic acid. The possibility of HSGA should be considered when an adenocarcinoma producing human salivary alpha-amylase is diagnosed away from sites where major and minor salivary glands normally are found.

Topics: Adenocarcinoma; Aged; alpha-Amylases; Carcinoembryonic Antigen; Choristoma; Diagnosis, Differential; Fatal Outcome; Head and Neck Neoplasms; Humans; Keratins; Male; Mucin-1; Neck; Neoplasms, Unknown Primary; RNA, Messenger; Salivary Gland Neoplasms; Salivary Glands

It has been demonstrated that CYFRA 21-1 (ELISA kit), which recognizes the soluble cytokeratin 19 fragment, is useful for assessing circulating tumor antigens in sera of patients with lung cancer. In this study, we compared the clinical significance of this new marker with the established squamous cell carcinoma antigen (SCC Ag), using sera from patients with head and neck malignant disease, healthy controls, and supernatants of established cell lines derived from squamous cell carcinomas and adenocarcinomas. The subjects were: Group A, 39 patients with malignant disease of the head and neck. Group B, 11 patients considered to be tumor-free after treatment. Group C, 67 patients with benign disease or healthy volunteers. Culture supernatants: 11 cell lines established from squamous cell carcinomas and adenocarcinomas. Serum levels of CYFRA 21-1 and SCC Ag of group A were significantly higher than those of group C. This finding suggests that CYFRA 21-1 is useful as a tumor marker as well as SCC Ag. CYFRA 21-1 and SCC Ag levels of patients in group A at the early and progressive stages of disease were comparable to the levels in group C. Both tumor markers are therefore useful for diagnosis of in the early stage of cancer. We attempted to set a cut-off level of CYFRA 21-1. The sensitivity of CYFRA 21-1 is higher than that of SCC Ag, especially in patients in the early stage of the disease. This finding indicates that the CYFRA 21-1 is preferable to SCC Ag as a tumor marker for the diagnosis of patients even in the early stages of malignant disease. The levels of CYFRA 21-1 in culture supernatants derived from tumor cell lines are higher than those of SCC Ag in all cell lines. The levels of CYFRA 21-1 are measurable, with levels varying with the cell line. There appears to be no correlation between the level of CYFRA 21-1 and the character of the cell lines, but this issue remains to be further investigated.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Keratin-19; Keratins; Male; Middle Aged

Theories regarding the origin of lateral neck cysts (LNC) range from congenital branchial pouch abnormalities to acquired salivary gland inclusions within lymph nodes. We analyzed 97 LNC histologically and evaluated their cytokeratin (CK) profile in a search for their derivation. 77/97 LNC were located in soft tissues, 20/97 within lymph nodes. LNC of young patients and of recent symptomatic presentation in older patients were lined by respiratory epithelium with scant lymphoid tissue, with expression of "simple epithelial" CK in ciliated cells and bimodal expression of "simple" and "stratified-epithelial-type" CK in basal cells. In longer standing symptomatic LNC, respiratory epithelium alternated with transitional-type pseudostratified epithelium with intraepithelial Langerhans cells and lymphoid hyperplasia, or consisted exclusively of squamous epithelium. We propose that respiratory epithelium is the "native" epithelium of LNC and squamous metaplasia results from inflammation induced stem cell hyperplasia in respiratory epithelium, evidenced by co-expression of "simple" and "stratified-epithelial-type" CK in all cells of transitional-type pseudostratified epithelium, the early stage in metaplastic transformation. Respiratory epithelium predominates in early LNC, lines pharyngeal tonsils and the recessus tonsillo-tubalis, and is a minor constituent of palatine tonsil but is not present in salivary glands. None of the LNC contained dysplasia, atypia, or carcinoma, or were associated with a primary carcinoma of tonsils or head and neck. We demonstrate that LNC arise from developmental remnants (congenital) of the 2nd branchial pouch, which may lie dormant for many years. Symptomatic enlargement, squamous metaplasia and lymphoid hyperplasia ensue as a consequence of immunologic stimulation a development reflected in hyperplastic palatine tonsils.

Topics: Adolescent; Adult; Aged; Biomarkers, Tumor; Branchioma; Child; Child, Preschool; Epithelium; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Infant; Keratins; Male; Middle Aged; Models, Biological

Retinoids are natural and synthetic analogues of vitamin A and have proven activity in various types of cancer. As for head and neck squamous cell cancer (HNSCC), retinoids are especially active in leukoplakia and in preventing second primary cancers. The aim of this study was to assess the growth inhibiting activity of all-trans retinoic acid (all-trans RA) in a panel of six head and neck squamous cell cancer cell lines and to correlate this response to the mRNA expression of factors related to differentiation and receptor mediated signal transduction. Three lines showed minimal, two moderate and one strong growth inhibition after 72 h exposure to all-trans RA. Three lines with a dissimilar response were selected for further studies, the measurement of mRNA expression by northern blotting. It was found that neither the expression nor the induction of retinoic acid receptor (RAR)-alpha and -gamma and retinoic X receptor-alpha mRNA war related to sensitivity. The mRNA expression of RAR-beta was too low to be measured in the three cell lines. The most sensitive cell line was, however, the only one that expressed mRNA of squamous differentiation markers. These data suggest a relationship between the retinoid sensitivity profile and the degree of cellular differentiation.

Topics: Carcinoma, Squamous Cell; Cell Division; Cornified Envelope Proline-Rich Proteins; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Keratins; Membrane Proteins; Neoplasm Proteins; Proteins; Receptors, Retinoic Acid; RNA, Messenger; RNA, Neoplasm; Tretinoin; Tumor Cells, Cultured

The prognostic weight of histological and biological factors was compared with that of known clinical prognostic factors in a population of 108 consecutive previously untreated patients with head and neck squamous cell carcinoma. Parameters studied were: tumour vascularisation, mitotic index, histological differentiation, nuclear grade, keratinisation, desmoplasia, growth pattern, inflammation, tumour emboli in peripheral vessels, keratins 6, 13, 19 immunohistochemical expression, cytofluorometric ploidy and S-phase. In multivariate analysis (Cox), only age and nodal status had a significant impact on the overall survival, whereas T stage was the only significant factor associated with locoregional failure. The cumulative incidence of metastases was correlated not only with age, T and N stage, but also with histological differentiation. All the other histological and biological factors studied failed to provide further prognostic information. These findings may help to select patients with high metastatic risk.

Topics: Age Factors; Aged; Analysis of Variance; Biopsy; Carcinoma, Squamous Cell; Confidence Intervals; Disease-Free Survival; Female; Flow Cytometry; Follow-Up Studies; Head and Neck Neoplasms; Humans; Inflammation; Keratins; Male; Middle Aged; Mitotic Index; Multivariate Analysis; Ploidies; Predictive Value of Tests; Prognosis; Prospective Studies; Retrospective Studies; S Phase; Survival Rate; Time Factors; Treatment Outcome

Retinoic acid (RA) modulates the growth and differentiation of various normal and malignant cells. These effects are most likely mediated by changes in gene expression. Genes whose expression is modulated by RA may be useful as markers of growth responsiveness to retinoids. Using differential cDNA cloning we identified 10 genes regulated by RA in the head and neck squamous cell carcinoma cell line MDA886Ln. Keratin (K) 13 gene expression was the gene expression most related to the degree of sensitivity of growth to RA, as K13 was not expressed in a series of RA-resistant cell lines. Our data suggest that low K13 expression may be mechanistically related to resistance to RA-induced growth inhibition.

Topics: Carcinoma, Squamous Cell; Cell Division; DNA, Complementary; Epithelium; Gene Expression; Head and Neck Neoplasms; Humans; Keratins; RNA, Messenger; Tretinoin; Tumor Cells, Cultured

Cytokeratin (CK) expression was studied in squamous cell carcinomas of different subsites in the head and neck by using cryostat sections from 27 head and neck squamous cell carcinomas (HNSCCs) and 6 cell lines established from HNSCC. All tissues were analyzed immunohistochemically with a panel of monospecific anti-keratin monoclonal antibodies. Most carcinomas recapitulated the expression pattern of keratins present in the basal layer of normal epithelium from the site of tumor origin. Regional differences in the expression of simple-epithelial type of keratins in stratified (pseudostratified) epithelia were to a large extent repeated in corresponding carcinomas. In the present study, localization of various keratins were surveyed and CK 18 specific monoclonal antibodies were specifically used to distinguish SCCs of the larynx or hypopharynx from SCCs of the oral cavity. CK 18 staining of almost all tumor cells was detected in 11 of 12 SCCs of the larynx and hypopharynx, but was only detected sporadically in 3 of 9 SCCs of the oral cavity. The present results show that CK 18 typing might be useful for distinguishing sites of origin of various HNSCCs. Findings also indicate that CK 18 expression in SCC might be modulated by microenvironmental factors.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Cell Line; Diagnosis, Differential; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Hypopharyngeal Neoplasms; Hypopharynx; Immunoenzyme Techniques; Keratins; Laryngeal Neoplasms; Larynx; Mouth Mucosa; Mouth Neoplasms

Patients with head and neck squamous cell carcinoma (HNSCC) who are thought to be cured are at high risk of development of a secondary primary tumour in the mucosa of the upper aerodigestive tract and the lungs. This phenomenon is in agreement with the concept of 'field cancerization', which implies that the whole mucosa is potentially condemned to the development of neoplasia. The hypothesis advanced in this study was that early markers of carcinogenesis should therefore be present in all cells of the mucosa of patients with HNSCC. The expression of cytokeratin 16, cytokeratin 19, and histo-blood group antigen H (ABH), type 2 chain was analysed by means of immunocytochemistry on exfoliated cells taken from six sites of the upper aerodigestive tract of the 'healthy' mucosa of previously untreated HNSCC patients (n = 25) and controls (n = 10). Statistically significant differences were found in the mucosal expression of these markers between patients and controls. Since no overlap in ABH type 2 chain expression existed between patients and controls and the expression between sites in a given individual was highly correlated, this marker was considered the most promising of those tested. These data suggest that cytokeratin 16, cytokeratin 19, and ABH type 2 chain are markers of field cancerization in easily available exfoliated cells, which may be applied to monitor and/or predict the occurrence of second primary tumours.

Topics: ABO Blood-Group System; Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mucous Membrane; Neoplasms, Second Primary

We investigated the incidence of micrometastases from squamous cell carcinomas of the head and neck in neck dissection specimens originally staged as pNO. A total of 76 dissection specimens from 60 patients were evaluated using serial microscopic sectioning in 10-microns intervals, H & E staining, and immunostaining with an antibody to pan-cytokeratin. Examination of 1,020 lymph nodes from 76 neck dissection specimens revealed 8 micrometastases (7.9%) in 6 specimens from 6 patients with oral and pharyngeal primaries, resulting in upstaging. Six micrometastases were located in lymph nodes 3 to 6 mm in diameter. The surgeon should be aware of the relatively high incidence of micrometastases from oral and pharyngeal carcinomas, which are undetectable preoperatively or by routine histopathologic examination. Primary tumor site (oral cavity and pharynx) and certain features of the primary can delineate a group of patients with a higher risk of harboring occult metastases who may benefit from elective treatment of the neck.

Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Histological Techniques; Humans; Immunohistochemistry; Keratins; Laryngeal Neoplasms; Lymph Node Excision; Lymphatic Metastasis; Mouth Neoplasms; Neoplasm Staging; Pharyngeal Neoplasms; Prognosis; Retrospective Studies

The authors performed a retrospective study in ultrasound to investigate new aspects in the sonomorphology of lymph node metastases of the neck. In this study, it could be demonstrated the first time that the histologic characteristics of the metastases determine the sonographic appearance. In addition to criteria such as the longitudinal/ transversal quotient, sonomorphology could support a more precise differential diagnosis of neck lymph nodes.. In 105 of 145 patients with histologically proved head and neck carcinomas, 187 lymph node metastases were detected by ultrasound. Sonomorphology was compared with the corresponding histology.. Five sonomorphologic groups could be differentiated. (1) Thirty-one percent of the metastases were homogenous. (2) Concerning the more complex morphology of lymph node metastases in ultrasound, echolucent forms could be differentiated from echogenic textures: low- or nondifferentiated and nonkeratinizing metastases appeared echolucent and cyst-like, with dorsal signal amplification. (3) Nonkeratinizing lymphomas with necrosis showed single or multiple echolucent intranodal lesions. (4) In correlation with an increasing keratinization, the echogenecity of the lymph nodes increased and intranodal echogenic inclusions appeared. (5) An extended keratinization correlated with a central echogenecity.. The morphologic assessment of lymph nodes in ultrasound allows for primary histologic and prognostic evaluation of lymph node metastases.

Topics: Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Neck; Necrosis; Ultrasonography

To determine the feasibility of using cytokeratin antibodies to distinguish normal and malignant cells in human tumors using flow cytometry. The goal was ultimately to increase the accuracy of cell kinetic measurements on human tumor biopsies.. A panel of four antibodies was screened on a series of 48 tumors from two centres; 22 head and neck tumors (Amsterdam) and 26 esophagus carcinomas (Leuven). First, screening was carried out by immunohistochemistry on frozen sections to test intensity of staining and the fraction of cytokeratin-positive tumor cells. The antibody showing the most positive staining was then used for flow cytometry on the same tumor.. The two broadest spectrum antibodies (AE1/AE3, E3/C4) showed overall the best results with immunohistochemical staining, being positive in over 95% of tumors. Good cell suspensions for DNA flow cytometry could be made from frozen material by a mechanical method, whereas enzymatic methods with trypsin or collagenase were judged failures in almost all cases. From fresh material, both collagenase and trypsin produced good suspensions for flow cytometry, although the fraction of tumor cells, judged by proportion aneuploid cells, was markedly higher for trypsin. Using the best cytokeratin antibody for each tumor, two parameter flow cytometry was done (cytokeratin versus DNA content). Enrichment of tumor cells was then tested by measuring the fraction of aneuploid cells (the presumed malignant population) of cytokeratin-positive cells versus all cells. An enrichment factor ranging between 0 (no enrichment) and 1 (perfect enrichment, tumor cells only) was then calculated. The average enrichment was 0.60 for head and neck tumors and 0.59 for esophagus tumors.. We conclude that this method can substantially enrich the proportion of tumor cells in biopsies from carcinomas. Application of this method could significantly enhance accuracy of tumor cell kinetic measurements.

Topics: Aneuploidy; Antibodies, Monoclonal; Biopsy; Carcinoma, Squamous Cell; Cell Cycle; DNA, Neoplasm; Esophageal Neoplasms; Feasibility Studies; Flow Cytometry; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins

Tumor of the follicular infundibulum is a rare proliferation of cells and its histogenesis or differentiation at the morphological level has been the subject of some controversy. In recent years cytokeratins have been recognized as important markers of epithelial differentiation, and of late new retrieval methods have meant it is possible to detect them in formalin-fixed and paraffin-embedded tissue. Four patients were studied, the ages ranging between the 2nd and 7th decade. The tumors were all located in the head and neck and in one case the lesion developed in a pre-existing sebaceous nevus. The morphological investigation revealed a flat, proliferation of polygonal, pale eosinophilic cells connected to epidermis or follicular infundibulum and with a centrally located nucleus. In addition, a few ductal structures resembling sebaceous ducts were seen and in one case a hair germ papilla and a follicular papilla was noted. Immunohistochemical investigations with antibodies against cytokeratins revealed differentiation comparable to that in the fetal follicular isthmus and, in one case, also differentiation in keeping with the fetal follicular infundibulum.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Basal Cell; Cell Transformation, Neoplastic; Cytoplasm; Female; Hair Follicle; Hamartoma; Head and Neck Neoplasms; Humans; Keratinocytes; Keratins; Male; Middle Aged; Precancerous Conditions; Sebaceous Glands; Skin Neoplasms

This study was designed to investigate the presence of nitric oxide in human squamous cell carcinoma of the head and neck. We localized the activity of nitric oxide synthase in these tumors through immunohistochemical analysis using antibodies to L-citrulline (a byproduct of nitric oxide synthase), to inducible nitric oxide synthase, and to constitutive nitric oxide synthase. We found presence of inducible enzyme in squamous cells throughout these tumors, with the highest intensity staining occurring directly around keratin pearls. Our findings suggest that inducible nitric oxide synthase activity is present in squamous cell carcinomas of the head and neck, leading us to conclude that inducible nitric oxide synthase may play a significant role in tumor growth.

Topics: Awards and Prizes; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Mouth Mucosa; Nitric Oxide; Nitric Oxide Synthase

Evaluation of Cyfra 21-1 (cytokeratin fraction 21-1) in squamous cell carcinoma of the head and neck.. Prospective study.. Serum Cyfra 21-1 concentration was measured in 250 samples from patients with squamous cell carcinoma of head and neck, patients with benign tumors of head and neck, healthy control subjects, and patients in remission from squamous cell carcinoma of head and neck.. Cyfra 21-1 concentration was elevated in 60% of the new patients with squamous cell carcinoma but only in 8% of patients with benign tumors and 3.5% of the healthy controls. At a cutoff of 1.3 ng/mL, the sensitivity of the test was 60%, the specificity was 94%, positive predictive value was 75%, and negative predictive value was 89%. The marker levels tended to follow the clinical course of the disease and were useful for therapy monitoring. Cyfra 21-1 levels were in good correlation with the tumor stage expressed by the local (T) and the lymphatic spread (N) and were inversely correlated with histologic grade, eg, higher in poorly differentiated carcinoma than in well-differentiated squamous cell carcinoma.. Cyfra 21-1 evaluation in head and neck squamous cell carcinoma is worthwhile for performance of an ample study that will prove and establish its routine use.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity

In head and neck squamous cell carcinoma a reliable serum marker of carcinogenesis should be of predictive value for the development of recurrent disease or a second primary tumour. At the moment, such a tumour marker is not available. Recently, elevated levels of cytokeratin 19-fragments (Cyfra 21-1) have been detected in the serum of patients with lung cancer, in particular with squamous cell carcinoma. Cytokeratin 19 is an intermediate cell filament protein expressed in simple epithelia and their malignant counterparts. Therefore, in this prospective study, a standardized sandwich enzyme-linked immunosorbent assay for soluble cytokeratin 19 fragments was tested in the serum of 20 patients with a previously untreated head and neck squamous cell carcinoma. The results were compared with that of 20 control individuals. Our results showed significantly higher Cyfra 21-1 concentrations in the serum of patients with cancer (10.21 +/- 3.03 ng/ml) than the controls (7.2 +/- 2.63 ng/ml). After radical treatment the marker levels dropped significantly to 1.65 +/- 1.07 ng/ml. Cyfra 21-1 appears to be of value as a circulating tumour marker for head and neck squamous cell carcinoma especially in monitoring disease control.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Enzyme-Linked Immunosorbent Assay; Epithelium; Female; Follow-Up Studies; Forecasting; Gene Expression Regulation, Neoplastic; Head and Neck Neoplasms; Humans; Keratins; Linear Models; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Neoplasms, Second Primary; Peptide Fragments; Prospective Studies

Malignant transformation is often associated with alterations in the expression of normal differentiation markers, which may serve as intermediate end points in carcinogenesis and cancer prevention. To identify early changes in differentiation markers during head and neck cancer development, we examined the expression of cytokeratins (CK1, CK8, CK13, and CK19) and involucrin by immuohistochemical methods in surgical specimens from 29 head and neck squamous cell carcinoma patients that included, in addition to carcinoma, adjacent dysplastic lesions (17 cases), hyperplastic lesions (21 cases), and adjacent histologically normal tissues (15 cases) and in specimens from 31 subjects with premalignant oral lesions (e.g, oral leukoplakia) without cancer, CK13 and involucrin were detected in all specimens from the cancer patients, and no differences in their expression were found among the different histopathological group. CK8 was detected in only 2.7% (1 of 36) of adjacent normal and hyperplastic tissues but in 58.8% (10 of 17) and 75.9% (22 of 29) of dysplastic and carcinoma tissues. The corresponding figures for CK19 expression in adjacent normal, hyperplastic, dysplastic, and carcinoma tissues were 13.3, 70, 71.4, and 82.1%, respectively. The expression of CK1 was not related to the progression from normal to malignant. In the leukoplakia lesions, CK8 CK13, CK19 and involucrin were detected in 13.8, 100, 74.2 and 100% of the specimens, respectively. These results demonstrate that CK19 expression increases in hyperplastic lesions and continues to be expressed in dyplastic and malignant lesions, whereas CK8 expression in low in adjacent normal and hyperplastic tissues and increases only in dysplastic and malignant lesions. Thus, CK19 and CK8 could be markers of sequential premalignant changes in head and neck carcinogenesis.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Culture Techniques; Disease Progression; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Sensitivity and Specificity

Since in vitro derived tumor cell lines usually correspond to their tumors of origin, a potential biological difference between a primary tumor and its derivative metastases and recurrent tumors should be reflected in established tumor cell lines. The aim of this study was to determine useful cellular markers in permanent tumor cell lines of head and neck squamous cell carcinomas (SCC) and to evaluate a possible relationship between these markers and the origin of selected cell lines. The cell lines, established in the laboratory of T. Carey at the University of Michigan (UM) (Ann Arbor, Mich., USA), were derived from primary tumor and its metastases (UM-SCC 10A, 10B), primary tumor and its recurrent tumors (UM-SCC 14A, 14B, 14C) and single tumors (UM-SCC 11B, 17A, 22B). An additional tumor cell line (HLac 79) was isolated by H.-P. Zenner (Tubingen, Germany) and a clone (8029 NA) with its cisplatin-resistant subline (8029 DDP4) was established in our laboratory. As markers we chose three groups known to be related to growth behavior and/or tumor differentiation: cytoskeletal proteins, oncogene products and membrane-associated antigens. These markers were detected by immunohistochemical methods using commercially available monoclonal antibodies. The "metastatic" and "recurrent" cell lines showed changes in comparison to the corresponding "parental" lines, which could be associated with a higher degree of de-differentiation, such as the occurrence of vimentin and neuroectodermal proteins, loss of HLA-ABC or HLA-DR and increased expression of epidermal growth factor receptor. The expression of cytokeratins was more stable and dissociation of the classical cytokeratin pairs was observed only in a few cases. Oncogene products were practically identical in cell lines from parental and recurrent or metastatic tumors. These data serve not only as a basis for further experiments with these cell lines but also provide information about the biological significance of various markers in newly established cell lines from primary tumors.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cisplatin; Clone Cells; Cytoskeletal Proteins; Drug Resistance, Neoplasm; ErbB Receptors; Head and Neck Neoplasms; HLA Antigens; HLA-A Antigens; HLA-DR Antigens; Humans; Immunohistochemistry; Keratins; Neoplasm Recurrence, Local; Oncogene Proteins; Pilot Projects; Tumor Cells, Cultured; Vimentin

The accumulation of keratinizing epithelium in the middle ear cavity is a crucial factor in the pathogenesis of cholesteatoma. We hypothesize that keratinocytes from the skin of the ear canal migrate and hyperproliferate in response to inflammation in the middle ear cavity to cause accumulation of keratin debris. In the present study, we investigated the expression of specific cytokeratins (CKs) in the cholesteatoma matrix to determine whether cholesteatoma is a hyperproliferative disease. Cytokeratin expression was examined in cholesteatoma, meatal skin, and tympanic membrane with two monoclonal antibodies, one for both cytokeratins 13 and 16 (antibody K8.12), and another for cytokeratin 13 only (antibody KS-1A3). CK 13 (MW 51 KD) is a marker of differentiation and CK 16 (MW 48 KD) is a marker of hyperproliferation of keratinocytes. The use of immunoblot probes showed that CKs 13 and 16 were present in cholesteatoma. Immunofluorescent staining showed the presence of CK 16 in the suprabasal layer of cholesteatoma, which was located near the external ear canal. CK 13 was localized in the suprabasal layer of meatal skin and tympanic membrane. CK 13 was localized in the basal layer of the cholesteatoma, distal to the external ear canal, but not in the meatal skin and tympanic membrane. Taken together, the present data suggest that cholesteatoma is a hyperproliferative disease and that cholesteatoma expresses CK 16 near the external ear canal and transforms to express CK 13 during growth distally.

Topics: Cholesteatoma; Ear Canal; Ear Diseases; Ear, Middle; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Gene Expression; Head and Neck Neoplasms; Humans; Immunoblotting; Keratins; Skin; Sodium Dodecyl Sulfate; Tympanic Membrane

Individual disseminated epithelial tumour cells were detected in bone marrow aspirates in 41 of 108 patients (37%) with squamous cell cancer of the head and neck region by an immunocytochemical technique based on monoclonal antibodies raised against the cytokeratin No. 19. In the clinical stage I (T1N0M0) tumour cells were detected only in 26.3% of the patients, whereas in stage IV (T4N0M0, T(all)N2-3M0, T(all)N(all)M1) almost twice as many patients (47.7%) presented with tumour cells in the bone marrow. Apparently, grade of differentiation of the tumour (grading) had no influence on the spread of single tumour cells. An influence of the different localisations of the primary tumour on tumour cell spread or the rate of tumour recurrence cannot as yet be discovered. Cytokeratin No. 19 expressing cells were not detectable in the bone marrow of 18 patients with non-malignant disease. Seventy-three patients were included in a follow-up study with a mean observation time of 25 months (range: 4-52 months). The presence of epithelial cells at the time of primary treatment appears to indicate a significantly higher risk of development of local or distant tumour recurrences (p = 0.01). Of 46 patients initially exhibiting no tumour cells in the bone marrow, only 14 had a clinical recurrence. Whereas 17 of 27 patients who presented with tumour cells in the bone marrow developed either a local tumour recurrence or distant metastases in different organs. Patients presenting with bone marrow tumour cells showed a significantly shorter disease-free survival than those without (p = 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Biomarkers, Tumor; Biopsy, Needle; Bone Marrow; Bone Neoplasms; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Leukocyte Common Antigens; Neoplasm Recurrence, Local; Prognosis

In a comparison of flow cytometric DNA measurements on fresh and paraffin-embedded material from primary squamous cell carcinomas of the head and neck region, we discovered that previously undetected aneuploid clones could be detected by dual parameter analysis of cytokeratin and DNA applied to disintegrated cells from paraffin sections. Using this new approach the correlation coefficient between DNA-indices from fresh and paraffin-embedded material increased from 0.423 to 0.904.

Topics: Aneuploidy; Carcinoma, Squamous Cell; DNA, Neoplasm; Epithelium; Fixatives; Flow Cytometry; Formaldehyde; G1 Phase; Head and Neck Neoplasms; Humans; Keratins; Paraffin Embedding; Resting Phase, Cell Cycle

The ability of all-trans-retinoic acid (RA) to modulate the growth and squamous differentiation of four head and neck squamous cell carcinoma cell lines (183, 886, 1483, and SqCC/Y1) was examined, and the relationship of their state of squamous differentiation and RA responsiveness to the expression of cytosolic RA-binding proteins (CRABPs), nuclear RA receptors (RARs), and retinoid X receptors (RXRs) was investigated. RA inhibited proliferation of all but the 183 cell line and suppressed squamous differentiation markers K1 keratin, type 1 transglutaminase, and involucrin mRNAs and proteins to varying degrees in 183, 1483, and SqCC/Y1 cells. Traces of CRABP-I mRNA were detected only in the 886 cells, whereas CRABP-II mRNA was detected in the other three cell lines. RA suppressed CRABP-II expression in SqCC/Y1 cells but had no effect on its expression in the other cell lines. All cell lines expressed mRNAs for RAR-alpha, RAR-beta, RAR-gamma, and RXR-alpha. The RAR-beta mRNA level was lowest in the SqCC/Y1 cells, and RXR-beta and RXR-gamma were not detected in any of the cell lines. RA treatment increased the levels of the three RAR mRNAs in most of the cell lines but had no effect on the RXR mRNAs. The CRABP-II mRNA level in SqCC/Y1 cells was lowest in cells grown in serum-free medium and increased when the cells were grown in medium with 5 or 10% serum. In contrast, the RXR-alpha mRNA level was inversely related to serum concentration. The results show that, in head and neck squamous cell carcinoma cells, there are no simple relationships among the expression of CRABPs, RARs, and RXRs and either squamous differentiation or response to RA-induced growth inhibition or suppression of squamous differentiation.

Topics: Carcinoma, Squamous Cell; Cell Differentiation; Head and Neck Neoplasms; Humans; Keratins; Protein Precursors; Receptors, Retinoic Acid; RNA, Messenger; Transglutaminases; Tretinoin; Tumor Cells, Cultured

One hundred forty-one head and neck squamous cell carcinomas were analyzed for keratin (K) 6, 13 and 19. Staining was evaluated by light microscopy (with or without grading) and image analyzer and expressed as a percentage of positive versus all tumor surface (PSA). Both techniques rendered strongly correlated results. Strong expression was noted in 108 carcinomas (76.1%) for K6, in 18 (12.7%) for K19 and in 21 (14.8%) for K13 (P = .001). One hundred thirty-six (96%) tumors were positive for K6, and their PSA ranged from 0.6% to 48.8%; K19, 48 cases (0.2-44%); K13, 59 (0.2-38.1%). Expression of K6 was related to differentiation. K19 was expressed mainly in moderately and poorly differentiated tumors, and K13 was manifest more in well-differentiated carcinomas or in keratinized areas of less-differentiated ones. Nineteen (13.38%) tumors were positive for both K13 and K19. K19 thus was related to tumor progression and K13 to differentiation. There was no correlation with tumor site or TNM category.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Discriminant Analysis; Female; Gene Expression; Head and Neck Neoplasms; Humans; Image Processing, Computer-Assisted; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplasm Metastasis; Prognosis

Cytokeratins are cytoskeletal components and constitute the intermediate filaments of epithelial cells. They are twenty in number and their distribution characterizes a very specific profile in each kind of epithelium. The authors characterized the cytokeratin repartition of the normal oropharyngeal epithelia in order to study their alterations in pathologic tissues, especially in neoplastic and dysplastic epithelia. The normal oropharyngeal epithelium shows cytokeratin pattern of non keratinized stratified epithelia. Three mucosa samples were studied from inflammatory, dysplastic and neoplastic epithelia. According to the alterations of cytokeratin repartition in the two last samples, cytokeratin pattern analysis could allow a characterization or the differentiation stage of neoplastic tissues before the expression of morphogenic criteria.

Topics: Antibodies, Monoclonal; Epithelium; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Oropharynx

As a basic study, an analysis was made, using flow cytometry (FCM), of a mixture of KB cells and lymphocytes after Propidium Iodide (PI)- Fluorescein Isothiocyanate (FITC) double strain. As a result, DNA histogram of only KB cells could be drawn. As a clinical application an analysis was made, using FCM, of tumor of the head and neck after double stain. It enabled the drawing of DNA histogram of only epithelial cells (cancer cells). Changes in DNA histogram of keratin positive cells were classified into three types: Type I: the same diploid type as whole cells, Type II: the same aneuploid type as them, and Type III: the different aneuploid type as them. Among 22 cases of malignant tumor there were 3 cases of Type I, 6 cases of Type II, and 13 cases of Type III. This method could contribute greatly to the detection of cancer cells and DNA diagnosis, and thus be useful clinically.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; DNA, Neoplasm; Epithelium; Fibroblasts; Flow Cytometry; Head and Neck Neoplasms; Humans; Keratins; Leukocyte Count; Lymphocytes; Ploidies

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

Topics: Antineoplastic Combined Chemotherapy Protocols; Biopsy, Needle; Carcinoma, Squamous Cell; Cobalt Radioisotopes; Cytoplasm; Diagnosis, Differential; Fluorouracil; Follow-Up Studies; Head and Neck Neoplasms; Humans; Keratins; Mitomycin; Neoplasm Recurrence, Local; Prospective Studies; Radiotherapy, High-Energy

Branchial cleft cysts were previously thought not to occur in the thyroid but have recently been described in two patients with Hashimoto's disease. This case report describes a patient with a branchial cleft cyst in an otherwise normal thyroid gland and could provide further evidence that thyroidal follicular cells are derived from the branchial clefts as well as from the primitive gut.

Topics: Adult; Branchioma; Epithelium; Female; Head and Neck Neoplasms; Humans; Keratins; Thyroglobulin; Thyroid Neoplasms

In this study of 40 cases of basaloid squamous cell carcinoma, 83% arose in the pyriform sinus, base of tongue, tonsil, and larynx. The 35 men and five women ranged in age from 27 to 88 years (median 62). In patients for whom social habits were recorded, 24 of 26 patients were smokers and 22 of 25 drank ethanol. Most presented with stage III or IV disease. Twenty-seven patients had regional metastases at the time of presentation and 15 developed distant metastases. Seventeen patients died with disease (median survival 18 months). The tumors were composed of moderately pleomorphic basaloid cells forming nests, cords, and frequent cribriform patterns. Squamous dysplasia of surface mucosa, focal squamous differentiation within invasive basaloid squamous cell carcinoma, or foci of conventional squamous cell carcinoma were present, alone or in combination. All studied neoplasms were immunohistochemically positive for keratins with the 34 beta E12 antibody. Approximately 80% were immunoreactive using AE1/AE3 or CAM 5.2. Epithelial membrane antigen, carcinoembryonic antigen, and S100 protein were found in 83%, 53%, and 39%, respectively, of the cases. Diffuse, weak immunoreactivity for neuron-specific enolase was seen in 75% of tumors. Synaptophysin, chromogranin, muscle-specific actin, and glial fibrillary acidic protein were absent. Basaloid squamous cell carcinoma has been confused with adenoid cystic carcinoma and small cell undifferentiated carcinoma, but is usually distinguishable in routine hematoxylin and eosin-stained sections, or, in rare problematic cases, with the aid of immunohistochemical studies. Distinction is warranted because the biologic behavior of basaloid squamous cell carcinoma differs from that of both of these lesions.

Topics: Adult; Aged; Aged, 80 and over; Carcinoembryonic Antigen; Carcinoma, Basosquamous; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; S100 Proteins

The incidence of micrometastases in cervical lymph nodes from squamous cell carcinomas of the head and neck was studied using routine histopathological examination. Micrometastases were found in 66 lymph nodes in 41 of the 92 tumor-positive neck dissection specimens. The detection of these micrometastases influenced postoperative treatment in 3 of the 77 patients with neck node metastases. The value of additional sectioning for detecting micrometastases was thus assessed. Sectioning at a deeper level in 600 originally histopathologically negative lymph nodes from 64 patients revealed 7 additional micrometastases in 5 patients. Antikeratin staining with a mixture of two monoclonal antibodies (AE1 and AE3) revealed 4 micrometastases in 739 originally histopathologically negative lymph nodes in 3 of 13 patients studied. Because of the unknown prognostic significance of micrometastases and the consequent arbitrary consequences for postoperative treatment, present findings show that the extra workload of immunostaining and deeper sectioning does not warrant their routine use in clinical practise.

Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunohistochemistry; Keratins; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Neck; Prognosis

Retinoids (vitamin A analogues) inhibit the squamous differentiation of normal and malignant epithelial cells. This study investigated the ability of the head-and-neck squamous-cell carcinoma (HNSCC) cell line 1483 to undergo squamous differentiation in the absence and presence of beta-all-trans retinoic acid (RA). The growth of these cells in culture is accompanied by an increase in keratinocyte transglutaminase, involucrin and keratin KI, 3 established markers of squamous cell differentiation. Higher levels of these differentiation markers were detected in cells cultured in delipidized serum (DLS), from which endogenous retinoids have been extracted, than in cells cultured in fetal bovine serum (FBS), which contains retinoids. Treatment with I microM RA decreased the levels of the various differentiation markers in cells cultured in either FBS or DLS as revealed by immunofluorescent labelling of permeabilized cells and by immunoblotting of cell extracts using specific monoclonal or polyclonal antibodies. The cells' ability to cross-link proteins to form envelopes under the plasma membrane was stimulated in the presence of calcium ionophore but inhibited by RA. These results indicate that the malignant 1,483 HNSCC cells recapitulate the main characteristics of normal squamous-cell differentiation in culture and that RA suppresses this differentiation as it does in normal keratinizing epithelial cells.

Topics: Calcium; Carcinoma, Squamous Cell; Cell Differentiation; Fluorescent Antibody Technique; Head and Neck Neoplasms; Humans; Immunoblotting; Ionophores; Keratinocytes; Keratins; Neoplasm Proteins; Protein Precursors; Retinoids; Transglutaminases; Tretinoin; Tumor Cells, Cultured

Keratin expressions in normal equine epidermis and experimentally induced equine papillomas were studied by immunohistochemical methods with three different human cytokeratin monoclonal antibodies, 34 beta B4 (directed against component 1), 34 beta E12 (directed against components 1, 5, 10, 11) and 35 beta H11 (directed against component 8). Staining patterns with 34 beta B4 and 34 beta E12 in the normal equine epidermis did not differ from those in the normal human epidermis. In the early developing papilloma, keratinocytes showed an abnormal suprabasal staining pattern and expressed an additional 56 kD keratin protein detected by 34 beta E12. In the advanced papilloma, cytolytic cells in the outer spinous and the granular layers did not stain positively with any of the three antibodies used. In both early and advanced papillomas, the expression of high molecular weight keratin proteins, as detected by 34 beta B4 and 34 beta E12, did not correlate with the degree of keratinization. By electron microscopy, keratinocytes in the advanced papilloma showed a marked decrease of tonofibrils and desmosome-tonofilament complex. These alterations may result from an abnormality in both proliferation and functional terminal differentiation of keratinocytes in the papilloma. There were obvious differences in staining patterns with 35 beta H11 between the normal human and equine epidermis; 54 kD keratin protein was expressed in suprabasal layers of the equine normal and papillomatous epidermis. Thus, this keratin protein may be regarded as a "permanent" marker for the equine epidermis.

Topics: Animals; Antibodies, Monoclonal; Epidermis; Head and Neck Neoplasms; Horse Diseases; Horses; Keratins; Microscopy, Electron; Molecular Weight; Neoplasm Proteins; Papilloma; Skin Neoplasms

Tissue samples taken from 22 patients before and during radical irradiation of squamous cell carcinomas in the head and neck region were studied by light and electron microscopy. The changes in keratinization pattern at the ultrastructural level seemed to be correlated with the outcome of the radiotherapy. The irradiation induced several cellular changes, of which nuclear atypia was the most prominent. This atypia was considered to be mainly due to cell death rather than to an aggressive nature of the tumor, because the number of mitoses decreased at the same time. The tumor invasion pattern remained unchanged. The keratinization pattern remained almost unchanged at the light microscopical level, but a slight increase of intracellular filaments and desmosomes was found in the electron microscopic study. The amount of intercellular filaments increased in three patients out of four with complete remission (CR), but in no case with tumor dissemination (n = 3) during radiotherapy. In patients with local persistent tumor or a local recurrence (LP + LR) (n = 15) the filaments either increased, decreased or remained unchanged. The number of desmosomes either increased or remained unchanged in three of four CR patients, in 13 of 15 LP + LR patients and in only one of three patients with tumor dissemination. They decreased in two patients with tumor dissemination, but only in one case with CR and in 2 cases with LP + LR. It is suggested that changes in cytoskeleton and desmosomes might be important in anchorage of tumor cells locally and might have value for prediction of the tumor response to radiotherapy. Further studies on larger materials are, however, needed before more definite conclusions can be drawn.

Topics: Carcinoma, Squamous Cell; Desmosomes; Epidermis; Head and Neck Neoplasms; Humans; Intermediate Filaments; Keratins; Microscopy, Electron

Twenty-five rhabdomyosarcomas (RMSs), including 12 alveolar and 13 embryonal types, were immunohistochemically studied for the presence of different classes of intermediate filament proteins and muscle actins (MAs). For the most part, formaldehyde-fixed and paraffin-embedded tissue was used in immunostaining. All RMSs showed desmin and MAs, usually in a major portion of tumor cells. The number of MA-positive cells was sometimes higher than that of desmin-positive cells. Vimentin was present in all tumors studied in frozen sections. Eight of 12 alveolar RMSs showed small number of cytokeratin-positive neoplastic cells. Cytokeratin-positive cells were present less commonly in embryonal RMS (3/13 cases). The 68-kD neurofilament protein was found in frozen sections of two embryonal RMSs. The cytokeratin and neurofilament immunostaining could be reproduced by immunofluorescence technique. In addition, we studied three childhood sarcomas, which showed abundant desmin and MA immunostaining but did not conform to the ultrastructural criteria of RMS. Scattered cytokeratin-positive cells were found in two of these tumors, and neurofilaments were found in the two cases for which frozen sections were available. The results show that typical RMS may demonstrate immunohistological pleomorphism with cytokeratin and neurofilament immunoreactivity suggesting the presence of multidirectional differentiation. In addition, there are tumors that by morphology look like RMS and have muscle cell markers but cannot be verified as RMS by electron microscopy; also, these tumors seem to show immunohistological pleomorphism. The presence of nonmyoid markers in RMS should be considered when making immunohistological diagnosis of soft tissue sarcomas.

Topics: Adolescent; Adult; Animals; Biomarkers, Tumor; Child; Child, Preschool; Female; Head and Neck Neoplasms; Humans; Immunohistochemistry; Infant; Intermediate Filament Proteins; Keratins; Male; Mice; Molecular Weight; Neurofilament Proteins; Prostatic Neoplasms; Rhabdomyosarcoma; Thigh

Tissue samples taken before and during the radical irradiation of the squamous cell carcinoma of the head and neck region were studied by light and electron microscopic examination. Radiation-induced cellular changes of which nuclear atypia was most pronounced. The tumor invasion pattern remained unchanged but the number of mitoses decreased. The lymphocytic infiltration increased at the beginning of the therapy (from 10-30 Gy) but decreased at the end of radiotherapy. The amount of neutrofils and the keratinization pattern remained almost unchanged at the light microscopic level, but intracellular filaments and desmosomes slightly increased in electron microscopic study. The changes in nuclear morphologic features pointing in a more undifferentiated direction are considered to be due to cell damage rather than to the more aggressive behavior of the tumor cells. This is in agreement with the decrease of mitoses which is due to radiation-induced arrest of tumor cells to the G2 phase. These changes might be related to the disappearance of tumors during irradiation. The leukocyte compartment seen in the samples might take part in the destruction of the tumor cells and in the removal of the cell debris.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cell Nucleus; Cytoskeleton; Desmosomes; Fibroblasts; Head and Neck Neoplasms; Humans; Keratins; Middle Aged

The anti-tumor activity of the putative differentiation-inducing agent dimethylformamide (DMF) was assessed in 7 head-and-neck xenograft (HNX) lines transplanted into nude mice. The drug was administered intra-peritoneally at the maximum tolerated dose. A significant growth-inhibitory effect was observed in 3 out of 7 tumor lines tested. When compared with 5 conventional drugs active in patients with squamous-cell carcinoma of the head and neck (HNSCC), DMF was as effective as the most active drugs (cisplatin and bleomycin). The most sensitive xenograft line, the poorly differentiated tumor HNX-14C, was used to test the hypothesis that differentiation induction might play a role in the anti-tumor activity of DMF. Light microscopic examination did not show clear-cut alteration of differentiation characteristics such as keratin and keratin pearl formation. Furthermore, we used a monoclonal antibody to study the expression of cytokeratin 10 which is useful as a differentiation marker of human HNSCC tumors. Keratin 10, not present in HNX-14C tumors grown under control conditions, became expressed in some cells upon DMF treatment. Further evidence for a differentiation-inducing activity of DMF was found in electron-microscopic studies. In treated HNX-14C tumors, in addition to cells with normal ultrastructural features, better-differentiated cells were observed, as manifested by an increase in the number of tonofilaments and desmosomes. The results show that DMF has a potential value for the treatment of patients with head-and-neck cancer, and that differentiation induction might play a role in the anti-tumor action of the drug.

Topics: Animals; Carcinoma, Squamous Cell; Cell Differentiation; Dimethylformamide; Drug Evaluation, Preclinical; Female; Head and Neck Neoplasms; Humans; Keratins; Mice; Mice, Nude; Microscopy, Electron; Neoplasm Transplantation; Tumor Cells, Cultured

Sixteen cases of malignant rhabdoid tumor (MRT) were studied by conventional light microscopy, immunohistochemistry, electron microscopy and flow cytometry. The age of the 16 patients varied from two months to 25.9 years. There were 11 males and five females. Eleven tumors were located in the kidney. The remaining five were found in the chest wall (n = 2) and the head and neck (n = 3). Particular histopathological findings included myxoid, pseudoalveolar and hyalinized areas. By immunohistochemistry, 15/15 cases stained positively for vimentin, 9/14 for cytokeratin, 6/15 for desmin, 9/14 for epithelial membrane antigen (EMA), 10/14 for neuron specific enolase (NSE) and 10/15 for protein S-100. Stains for neurofilaments, myoglobin and Ulex europaeus aggl. I (UEA I) were negative. The characteristic finding by electron microscopy in three cases were large numbers of intermediate filaments arranged either randomly or in concentric whorls. None of the 11 cases studied revealed aneuploid DNA stem lines as determined by flow cytometry. Of the 16 patients 12 died, one is living with disease and three are living without evidence of disease. Postoperative treatment consisted of chemotherapy, in some cases combined with radiotherapy. Two patients developed a medulloblastoma in addition to a renal and extrarenal MRT, respectively. Our findings demonstrate that MRT may present more histopathological patterns than hitherto recognized. In addition, they show that MRT may express a wide range of antigenic "markers", similar to epithelioid sarcoma with which it may be confused on cytological grounds. Despite aggressive postoperative chemotherapy prognosis is still poor.

Topics: Adolescent; Adult; Age Factors; Aneuploidy; Cell Nucleus; Child; Child, Preschool; Desmin; DNA; Female; Flow Cytometry; Head and Neck Neoplasms; Humans; Immunohistochemistry; Infant; Keratins; Kidney Neoplasms; Male; Microscopy, Electron; Phosphopyruvate Hydratase; Rhabdomyosarcoma; S100 Proteins; Sex Factors; Thoracic Neoplasms; Vimentin; Wilms Tumor

The recognition of myogenic differentiation is not always possible using routine light or electron microscopic techniques. In this article, we describe our experience with two small round-cell neoplasms occurring in young children, each of which exhibited an undifferentiated light microscopic appearance. In both cases, myogenic features were revealed by immunocytochemical methods applied to fine-needle aspiration (FNA) biopsies. Each was immunoreactive for desmin and vimentin and failed to react with antibodies to leukocyte-common antigen. Moreover, formalin-fixed tissue sections of one case were negative for cytokeratin and epithelial membrane antigen. Sporadic reactivity for neuron-specific enolase and Leu-7 antigen was observed in occasional cells in FNA specimens, but synaptophysin was not identified. The rapidity of the method and the reliability of the immunocytochemical findings, when compared with routine cytologic evaluation, emphasize the diagnostic utility of immunocytochemical analysis when FNAs of pediatric soft-tissue tumors are obtained.

Topics: Actins; Biopsy, Needle; Desmin; Diagnosis, Differential; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Infant; Keratins; Male; Membrane Glycoproteins; Mucin-1; Phosphopyruvate Hydratase; Rhabdomyosarcoma; S100 Proteins; Vimentin

Cell line MDA 886Ln was established from a laryngeal lymph node metastasis. When grown as a multicellular tumor spheroid (MTS), it exhibits squamous differentiation. We studied the effects of beta-all-trans retinoic acid (RA) on the growth, differentiation and glycoprotein content of this MTS model for squamous carcinomas of the head and neck. The growth of MTSs was inhibited in a dose-dependent manner by 10(-6) to 10(-10) M RA. Growth inhibition occurred between 3 and 5 days of RA treatment (10(-6)M). Immunohistochemical and electrophoretic analyses revealed that RA suppressed the morphological markers of squamous differentiation (squames), involucrin expression, and keratin expression. Gly-coprotein expression was examined by metabolic labelling using 3H-glucosamine, in situ labelling of polyacrylamide gels with 125I-labelled wheat-germ agglutinin (WGA), localization of fluorescein isothionate-WGA in frozen sections, and determination of sialyltransferase activity. Treatment using 10(-6) M RA altered glycoprotein expression both biochemically and morphologically, and WGA was shown to bind preferentially to sialic acid residues. The sensitivity of this MTS model to RA treatment and its ability to be analyzed through morphological, immunohistochemical and biochemical techniques suggest that it will prove useful in studying the relationships between growth, differentiation and RA-induced alterations in squamous carcinomas.

Topics: Carcinoma, Squamous Cell; Cell Differentiation; Cell Division; Glycoproteins; Head and Neck Neoplasms; Humans; Keratins; Male; Molecular Weight; Neoplasm Proteins; Organoids; Protein Precursors; Tretinoin; Tumor Cells, Cultured

It has been suggested that cytokeratin 13 is a useful marker of malignancy. We examined normal squamous cell epithelia, hyperplasia, dysplasias of various grades, intraepithelial neoplasia and invasive squamous cell carcinomas of the pyriform fossa using K13 and KL1. Positive staining for K13 was seen in all normal or hyperplastic benign epithelia, was inconstant in dysplasia, and intraepithelial neoplasia and carcinoma was negative. KL1 expression is constant and non significant. These results suggest that tumour cells are unable to synthesize keratin 13 a finding which may be valuable in surgical pathology.

Topics: Biomarkers, Tumor; Carcinoma; Carcinoma, Squamous Cell; Epithelium; Gene Expression; Head and Neck Neoplasms; Humans; Hyperplasia; Immunohistochemistry; Keratins; Larynx; Pharynx

Two human cell lines were established from untreated squamous cell carcinomas of the head and neck. Line 183 was derived from a head and neck squamous cell carcinoma of the tonsil and 1483 from a head and neck squamous cell carcinoma of the retromolar trigone. Both lines grow in a cobblestone pattern demonstrating their epithelial heritage. Immunofluorescence studies and one-dimensional polyacrylamide gel electrophoresis indicated that both lines contain cytokeratins. Line 1483 is more aggressive in nude mice, has a higher efficiency for anchorage-independent growth, expresses p21ras (product of the ras oncogene) at a higher level, and is more aneuploid than 183. 1483 also grows as a multicellular tumor spheroid. Line 1483, which was established from the primary tumor of a patient with nodal metastasis, thus displays more progressed characteristics than line 183, which was established from a patient with no clinically positive nodes.

Topics: Aged; Carcinoma, Squamous Cell; Cell Aggregation; Chromosome Aberrations; Genetic Markers; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Proto-Oncogene Proteins; Proto-Oncogene Proteins p21(ras); Tumor Cells, Cultured

In this study, the immunoreactivity of several cytokeratin antibodies; 115 D8, a monoclonal antibody against MAM-6, an epithelial membrane antigen; and two vimentin antibodies, is examined in relation to the degree of morphologic differentiation in carcinomas of the head and neck. The results indicate that a relationship exists between the degree of morphologic differentiation and the expression of cytokeratin, MAM-6, and vimentin, as detected by polyclonal antikeratin, 115 D8 and anti-vimentin. Expression of cytokeratin and MAM-6 is reversely related to vimentin. Polyclonal anti-keratin; CAM 5.2, a monoclonal antibody against cytokeratin 8, 18 and 19; and 115 D8, used in combination, were still able to identify the epithelial nature of undifferentiated/spindle cells. Since these immunohistochemical markers precede light microscopic detectable signs of epithelial differentiation, they can be used for the identification of the epithelial nature of undifferentiated/spindle tumors of the head and neck.

Topics: Antigens, Differentiation; Carcinoma; Carcinoma, Squamous Cell; Cytoskeleton; Desmin; Diagnosis, Differential; Head and Neck Neoplasms; Humans; Keratins; Membrane Glycoproteins; Microscopy, Electron; Mucin-1; Staining and Labeling; Vimentin

Twenty-nine paragangliomas of the head and neck region including 20 glomus jugulare (GJ) and nine carotid body (CB) tumors were evaluated for the presence of neuroendocrine and intermediate filament antigens. Immunohistochemistry on formalin-fixed, paraffin-embedded tissue was used to identify: S-100 protein (S-100); neuron-specific enolase (NSE); chromogranin A (CHA); serotonin (SER); synaptophysin (SYN); cytokeratin (CK); neurofilament (NF); desmin (DES); vimentin (VIM); and glial fibrillary acidic protein (GFAP). S-100 protein staining of sustentacular cell nuclei and cytoplasm was found in all tumors and was present in chief cells in 4 of 20 GJ and 3 of 9 CB tumors. All tumors stained with at least three neuroendocrine markers (29 of 29 NSE, 28 of 29 SYN, 26 of 29 CHA, 25 of 29 SER). CK was detected in 2 GJ and 1 CB tumor using anticytokeratins AE 1/3 and CAM 5.2. Neurofilament protein could not be demonstrated in fixed material, and all tumors were negative for GFAP and desmin. Vimentin was inconsistently detected in chief and sustentacular cells. We conclude that, in formalin-fixed material, paragangliomas have S-100 protein staining of sustentacular cells with chief cells containing antigens associated with neuroendocrine differentiation. The presence of CK in some paragangliomas is consistent with recent tissue culture studies demonstrating immunoblot confirmation of CK in pheochromocytomas and represents a potential source of immunohistologic misinterpretation in diagnosis, unless a panel of markers is utilized.

Topics: Antibodies; Chromogranin A; Chromogranins; Desmin; Glial Fibrillary Acidic Protein; Head and Neck Neoplasms; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Membrane Proteins; Neurofilament Proteins; Neurosecretory Systems; Paraganglioma; Phosphopyruvate Hydratase; S100 Proteins; Serotonin; Synaptophysin; Vimentin

Effective treatment modalities for Stage III and IV squamous cell carcinomas (SCC) of the head and neck are limited and seldom result in long term survival. The improved results with cisplatin containing chemotherapy have been encouraging and represent an additional therapeutic modality for head and neck cancer. To estimate the effectiveness of concomitant radiation and cisplatin chemotherapy, the Radiation Therapy Oncology Group (RTOG) initiated a Phase II study for patients with advanced nonresectable SCC of the head and neck. In addition, the diagnostic biopsy specimens were collected. Two pathologists reviewed and scored the biopsy specimens for a number of histologic parameters, including degree of keratinization, nuclear pleomorphism, frequency of mitoses, inflammatory and stromal reaction, pattern of invasion and vascular involvement in order to identify potential prognostically important patient subgroups. A total of 114 patients were evaluated for complete clinical responses (CR). These were achieved in 76% for all head and neck sites (ALL), and in 72% of the patients excluding nasopharyngeal and sinus cancers (REST). Evaluation of histopathologic parameters through multivariate analysis identified the presence of keratin as the most significant in predicting CR. Non-keratinizing SCC had CRs of 98% (ALL), and 94% (REST), as compared with 64% and 67% in the patients with keratin producing neoplasms (P less than 0.001 and 0.05) respectively. Survival at 24 months was found to be improved in the non-keratinizing SCC (P = 0.002). Multivariate analysis also identified the frequency of mitoses as being important in predicting for CR in patients with keratin in the biopsy findings. Biopsy specimens from ALL patients with two or more mitotic figures per high-power microscopic field had 76% CRs, in comparison with 46% when none or one mitotic figure was observed (P = 0.02). In the restricted group of patients (REST), the CR was 77% in the high mitotic figure group as opposed to 45% in the lower rate group (P = 0.03). However, this significant difference in CR rates did not translate into improved survival for the high CR subset.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Cisplatin; Combined Modality Therapy; Drug Evaluation; Female; Head and Neck Neoplasms; Histocytochemistry; Humans; Keratins; Male; Middle Aged; Neoplasm Staging; Statistics as Topic

Immunohistochemical analysis of 21 prototypic mucosal spindle-cell carcinomas of the aerodigestive tract was performed at the Armed Forces Institute of Pathology (AFIP) to establish the usefulness of selected immunohistochemical markers in distinguishing spindle-cell carcinoma from other mucosal spindle-cell neoplasms. Immunoreactive keratin could be demonstrated in only 13/21 (62%) of cases. Coexpression of keratin and vimentin was demonstrated in 10/17 (59%) of the tumors evaluated for both of these intermediate filaments. All spindle-cell carcinomas lacked S100 protein, which is an immunoreactivity we would expect to find in spindle-cell malignant melanoma, one of the principal considerations in a differential diagnosis. Both alpha-1-antitrypsin (AAT) and alpha-1-antichymotrypsin (ACT) were demonstrated in the tumor cells in all cases. However, albumin had a similar distribution in the tumors, which suggested that passive uptake was a serious confusing factor. The results of this study indicate that AAT and ACT are unreliable markers for distinguishing spindle-cell carcinomas from malignant fibrous histiocytomas.

Topics: Aged; alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Antigens, Neoplasm; Carcinoma; Female; Head and Neck Neoplasms; Humans; Keratins; Male; Middle Aged; Muramidase; S100 Proteins; Serum Albumin; Vimentin

The neuroendocrine carcinoma of the skin has its histogenetic origin in Merkel cells and a preference in head and neck area in the seventh decade of life. The definitive diagnosis can be made with a combination of electron microscopy and immunohistochemistry. Merkel cell carcinoma is a primary cutaneous neoplasma and is rarely found on the lips or gingiva. Operation and radiation are the therapy of choice. The value of an additional antineoplastic chemotherapy in the treatment of Merkel cell carcinoma is still controversial. Although long survival times had been described in literature the occurrence of local relapses and metastases demands for frequent controls.

Topics: Aged; Biopsy; Female; Head and Neck Neoplasms; Histocytochemistry; Humans; Intermediate Filament Proteins; Keratins; Lymph Node Excision; Lymphatic Metastasis; Microscopy, Electron; Neurofilament Proteins; Skin Neoplasms

The effects of butyrate and retinoic acid in combination with catecholamines or histamine on the HN-1 human head and neck squamous carcinoma cell line were investigated analysing cell proliferation, placental alkaline phosphatase (PLAP) activity, and relative cytokeratin content. Butyrate inhibited cell proliferation in agar, whereas retinoic acid induced a small inhibitory effect. Butyrate enhanced PLAP activity in a time related manner in contrast to retinoic acid, which had no significant effect. However, retinoic acid inhibited the efficacy of butyrate to induce PLAP activity. A synergistic enhancement of PLAP activity was demonstrated after treatment of butyrate pretreated cells with catecholamines or histamine. The beta-adrenergic antagonist propranolol partly inhibited the aforementioned enhancement of PLAP activity, whereas the alpha-adrenergic antagonist phentolamine further enhanced PLAP activity. Indirect labeling of keratins with a polyclonal antibody showed that cytokeratin content was enhanced by butyrate but not by retinoic acid. Further analysis of cytokeratin content using four monoclonal antibodies showed that labeling of cytokeratins (5 + 8) was increased by butyrate. PLAP activity could be modulated by a concerted action of either butyrate plus retinoic acid or butyrate plus catecholamines or histamine, indicating a possible role for PLAP in tumour cell proliferation.

Topics: Alkaline Phosphatase; Butyrates; Butyric Acid; Carcinoma, Squamous Cell; Catecholamines; Cell Division; Cell Line; DNA; Flow Cytometry; GPI-Linked Proteins; Head and Neck Neoplasms; Histamine; Humans; Isoenzymes; Keratins; Phentolamine; Placenta; Propranolol; Tretinoin

Keratins are alpha-type fibrous polypeptides which basically compose 10 nm thick or intermediate-sized filaments (IF) in almost all epithelial cells and tissues. Their patterns of expression in normal and malignant upper digestive tract squamous epithelium were monitored by high resolution gel electrophoresis, immunoblotting, and immunohistochemical techniques. Uninvolved epithelia, in all instances, were found to express keratins 4, 5, 6 and 13, 14 the members of the high molecular weight basic (type II or Type B) and of the low molecular weight acidic (type I or type A) subfraction, respectively. Cancers of squamous epithelial cell origin retain keratin synthesis. However, their overall patterns of keratin expression appeared aberrant when compared with those of normal epithelia. In particular, these differences result from highly proliferative tumour cells unable in most cases to synthesize keratins 4/13, a type II/type I keratin pair which specifically indicates in squamous primarily non-keratinizing epithelia completely, i.e. terminally differentiated (suprabasal or spinous) cells. The patchwise expression of acidic keratin 13 in related primaries confirms their heterogeneous phenotype, and may be explained, in part, by cancer cells no longer resistant to terminal differentiation as a result perhaps of an altered micro-environment and/or in response to various effects mediated by vitamin A. We discuss some problems pertinent to the biochemical analysis of keratin polypeptides in normal and involved epithelial tissues, and relate to the controversial question whether specific keratin members may actually candidate for markers of malignancy.

Topics: Carcinoma, Squamous Cell; Cell Division; Cell Transformation, Neoplastic; Epithelium; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Neoplasm Staging

Cell subsets have been discriminated in cell suspensions derived from 37 human head and neck tumors by means of light scatter, DNA, and cytokeratin flow cytometry (FCM). Cell dispersion was performed overnight at 4 degrees C in two different enzyme mixtures, i.e., trypsin/dithioerythritol and collagenase/DNase, under slight agitation of sliced tumor tissue. Cells were examined before and after fractionation on a discontinuous low-density bovine serum albumin (BSA) gradient. Forward and right-angle light scatter FCM of 23 tumor specimens revealed four main subpopulations with different size and structure. Fractionation of primary cell suspensions on a BSA gradient at unit gravity separated debris, small cells and large cells. DNA FCM of the enriched populations demonstrated a relation between large cells and DNA aneuploidy. Epithelial cells, as recognized by cytokeratin antibodies, were also related with large cells. The results demonstrated the usefulness of light scatter, DNA, and cytokeratin analysis of crude and fractionated tumor cell suspensions for assessment of the efficacy of a particular dispersion technique and to obtain information of the cell subsets dispersed.

Topics: Cell Separation; Centrifugation, Density Gradient; DNA, Neoplasm; Flow Cytometry; Head and Neck Neoplasms; Humans; Keratins; Scattering, Radiation; Serum Albumin, Bovine; Suspensions

Different tumours of the head and neck were analysed by immunohistochemistry. The distribution pattern of several intermediate filaments was studied. Keratin filaments were typical of carcinomas, whereas vimentin filaments were typical of mesenchymal tumours of different origin. The advances of this new technique of "tumour typing" are discussed.

Topics: Carcinoma, Basal Cell; Cytoskeleton; Diagnosis, Differential; Head and Neck Neoplasms; Humans; Hypopharyngeal Neoplasms; Immunoenzyme Techniques; Intermediate Filaments; Keratins; Lymphoma; Melanoma; Mouth Neoplasms; Nasopharyngeal Neoplasms; Neuroma, Acoustic; Oropharyngeal Neoplasms; Tonsillar Neoplasms; Vimentin

Topics: Antibodies; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Head and Neck Neoplasms; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins

The histogenetic origin of the spindle-cell component of spindle-cell carcinoma of the head and neck mucosa remains controversial. The spindle cells have been considered a variant growth pattern of squamous-cell carcinoma, a non-neoplastic mesenchymal reaction, and a malignant admixture of epithelial and mesenchymal neoplasm. To evaluate the spindle-cell component, we studied 25 tumors (18 biphasic and seven monophasic) by utilizing the following: an avidin-biotin complex immunoperoxidase technique with a variety of antikeratin antibodies (AE1, AE3, CAM 5.2, 35BH11, and polyclonal Dako) and a monoclonal antivimentin antibody, and an avidin-biotin alkaline phosphatase double-labeling technique to detect coexpression of keratin and vimentin. The immunohistologic staining pattern was compared with electron-microscopic studies. Eight of 18 biphasic neoplasms contained immunoreactive keratin in the spindle-cell component that was distributed focally in a minority of cells in 3 tumors and diffusely throughout five of the neoplasms. Four of seven ulcerated monophasic spindle-cell tumors devoid of histologic squamous-cell carcinoma also were keratin positive, confirming epithelial differentiation. The majority of the spindle cells in all the tumors contained vimentin intermediate filaments. In three immunoperoxidase keratin positive biphasic tumors examined with alkaline phosphatase double labeling, occasional spindle cells were found that coexpressed keratin and vimentin and were interspersed with cells expressing either intermediate filament. Electron microscopy was performed on the spindle-cell component of 13 tumors, nine biphasic and four monophasic. Of the biphasic tumors, four were immunoperoxidase keratin positive; three of these showed epithelial differentiation by electron microscopy. Five biphasic tumors were keratin negative, and three tumors had epithelial differentiation by electron microscopy. Four monophasic spindle-cell tumors were immunoperoxidase keratin positive, and one of these had epithelial features by electron microscopy. Two monophasic tumors were keratin negative and without ultrastructural evidence of epithelial features. By using a combination of immunohistochemical and electron-microscopic observations, we identified evidence for epithelial differentiation in the spindled cells in 11 of 18 biphasic tumors and four of seven monophasic spindle-cell tumors.

Topics: Aged; Aged, 80 and over; Antibodies, Monoclonal; Carcinoma; Carcinoma, Squamous Cell; Female; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Laryngeal Neoplasms; Male; Microscopy, Electron; Middle Aged; Mucous Membrane; Vimentin

Spindle cell squamous carcinoma is a highly malignant neoplasm, characterized by a biphasic appearance, due to the presence of a diffuse proliferative pattern of pleomorphic spindle cells with extremely rare foci of conventional squamous cell carcinoma. Epithelial markers in eight cases of spindle-cell squamous carcinoma of the head and neck region were studied by immunocytochemistry. Epithelial membrane antigen and cytokeratin resulted strongly positive in the spindle cells of the sarcoma-like areas demonstrating the epithelial nature of the neoplasm. In 6 of our patients who later died from widespread metastases, the neoplasm had poor prognosis with a mean survival rate of 14.7 months.

Topics: Antigens, Neoplasm; Carcinoma, Squamous Cell; Epithelium; Head and Neck Neoplasms; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Membrane Proteins; Mucin-1

Topics: Carcinoma, Squamous Cell; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Molecular Weight

Four human cell lines were established from biopsy specimens of squamous cell carcinomas of the larynx (TR131 and TR138), tongue (TR126), and buccal mucosa that had infiltrated a lymph node (TR146). All 4 lines readily formed colonies on a plastic substratum, but they were virtually incapable of forming colonies in an anchorage-independent semisolid support system of soft agar (cloning efficiencies, less than 0.02%). The proliferation of this group of tumor-derived cell lines, therefore, appeared to be highly anchorage dependent. Keratin filaments could be visualized in each line by indirect immunofluorescence with the use of polyclonal or monoclonal antibodies to keratins; staining with monospecific antibodies indicated that 3 of the 4 lines expressed simple epithelial keratins 8 and 18, whereas 1 of the 4 also expressed keratin 19. A panel of lectins revealed characteristic localization patterns distinct from those observed on other epithelial cell lines. Cells from 3 lines (TR131, TR138, and TR146) inoculated into nude mice (nu/nu) produced cystic nodules or unequivocal tumors having a histology indicating a squamous cell origin for the injected cells. Electron microscopy demonstrated that the cell lines covered a spectrum of differentiation capability ranging from the undifferentiated monolayer cultures of TR126 to the rather well differentiated, stratified cultures of TR131.

Topics: Adult; Aged; Animals; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line; Cytoskeleton; Female; Head and Neck Neoplasms; Humans; Keratins; Lectins; Male; Mice; Mice, Nude; Middle Aged; Neoplasm Transplantation; Transplantation, Heterologous

Immunohistochemical localization of keratin antigens using keratin antisera and the immunoperoxidase technique have been shown to be helpful in identifying certain epithelial cells. Our study was designed to evaluate the application of this technique to head and neck neoplasms and normal tissues using two keratin antibody preparations. Our data indicate that the keratin antibodies stained normal epithelial structures in the head and neck except for cells with active secretory functions such as mucus, cerumen, or salivary secretion. Neoplasms of the head and neck showed keratin antibody staining for epithelial neoplasms and negative staining for mesenchymal neoplasms. The immunohistologic demonstration of keratin is useful in distinguishing undifferentiated or poorly differentiated epithelial malignancies from sarcomas or lymphomas and demonstrating myo-epithelial cells in salivary neoplasms.

Topics: Animals; Antibodies; Ear; Head and Neck Neoplasms; Humans; Immunoenzyme Techniques; Keratins; Mice; Pharynx; Rabbits; Swine

With indirect immunofluorescence microscopy it is possible to visualize intermediate-sized filaments which show a cell-specific distribution and in this manner establish a light-microscopical diagnosis in certain cases which are difficult or impossible to differentiate using conventional methods. We applied the same method to tumours of the head and neck region. Intermediate-sized filaments were studied in four malignant lymphomas, seven carcinomas and four metastases of carcinomas. Malignant lymphomas showed a positive reaction with antibodies to vimentin, carcinomas a positive reaction with antibodies to keratin. Using a monoclonal antibody against a single keratin polypeptide (cytokeratin 18) a further subdivision of the carcinomas was possible. The keratinizing squamous cell carcinoma and two non-keratinizing squamous cell carcinomas showed a positive reaction with the conventional antibody against keratin, but a negative reaction with the monoclonal antibody against cytokeratin 18. One adenocarcinoma, two anaplastic carcinomas and one lymphoepithelial carcinoma were positive with the conventional antibody against keratin and with the monoclonal antibody against cytokeratin 18. Thus lymphoepithelial carcinomas and anaplastic carcinomas should probably not be regarded as special variants of squamous cell carcinoma. In all metastases the same intermediate-sized filaments were demonstrable as in the primary tumour. Certain advantages of immunofluorescence microscopy when compared to diagnostic electron microscopy are discussed.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoma; Carcinoma, Squamous Cell; Cytoskeleton; Diagnosis, Differential; Fluorescent Antibody Technique; Head and Neck Neoplasms; Humans; Keratins; Lymphatic Metastasis; Lymphoma, Non-Hodgkin; Microscopy, Electron; Vimentin

Immunohistochemical staining for keratin proteins may be useful as a diagnostic parameter in head and neck neoplasms. Our study evaluates the keratin antibody staining properties of normal tissues as well as neoplastic and non-neoplastic head and neck lesions from surgical procedures performed on 100 patients. The results indicate that the anti-keratin antibody technique can be helpful in several areas of head and neck pathology.

Topics: Antibodies; Antibodies, Neoplasm; Head; Head and Neck Neoplasms; Humans; Immune Sera; Immunoenzyme Techniques; Keratins; Nasal Mucosa; Pharynx

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Granuloma; Head and Neck Neoplasms; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Skin Neoplasms