bromochloroacetic-acid and Urinary-Bladder-Neoplasms

Small cell carcinoma of the urinary bladder is a rare, often fatal, disease. Its presenting symptoms and gross morphology are similar to those of conventional urothelial carcinoma, whereas its prognosis is much poorer with frequent metastasis. Small cell carcinoma of the urinary bladder shares similar histology with its counterparts in other organs; however, its immunoreactivity to conventional neuroendocrine markers is low. Its diagnosis is thus considered permissible on morphologic grounds alone. Multimodal treatments are often employed, although no definite treatment algorithm has been established. For this extremely aggressive malignancy with an as-yet inconclusive etiology, further studies are needed to clarify its molecular pathogenesis to serve as a basis for diagnostic markers and therapeutic targets. The clinical, morphologic, immunoreactive, molecular, and therapeutic features of bladder small cell carcinoma are reviewed, including a detailed discussion on the utility of immunohistochemical markers.

Topics: Biomarkers, Tumor; Carcinoma, Neuroendocrine; Carcinoma, Small Cell; Cyclin-Dependent Kinase Inhibitor p16; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Male; Molecular Biology; Nuclear Proteins; Prognosis; Thyroid Nuclear Factor 1; Transcription Factors; Urinary Bladder Neoplasms

Primary adenocarcinoma of urinary bladder is an uncommon neoplasm and is a source of diagnostic confusion with adenocarcinomas arising in adjacent organs, especially colon. These tumors show varied histologic picture and degree of differentiation. Clinical association with bladder exstrophy and schistosomiasis has been well documented. Primary bladder adenocarcinomas have overlapping histologic and immunohistochemical features with adenocarcinomas arising from other primary sites and the suggested immunohistochemical panel includes cytokeratins 7 and 20, 34βE12, thrombomodulin, CDX2, and β-catenin. Clinical, imaging, histologic, and immunohistochemical correlation should be done while rendering this diagnosis, as prognosis and therapeutic options for primary versus metastatic adenocarcinoma vary widely.

Topics: Adenocarcinoma; beta Catenin; CDX2 Transcription Factor; Homeodomain Proteins; Humans; Immunohistochemistry; Keratin-20; Keratin-7; Keratins; Microfilament Proteins; Thrombomodulin; Urinary Bladder Neoplasms

Finding and development of new bladder cancer markers is still a very dynamic field. Because of the mass of all these markers it is impossible to report all of them. This paper reviews the role of bladder cancer markers in diagnosis and highlights the most important biomarkers studied and reported recently. A medline based literature search was performed to examine the field of bladder cancer markers. Major topics focus on selected bladder cancer markers from nearly all categories of the wide field of bladder cancer markers: Hematuria, FISH, FGFR3, SURVIVIN, u-PAR, TP53 mutation, HER-2/neu, TPA, NMP22, CK-19, CK-20, CYFRA 21-1. The use and clinical importance as diagnostic help are discussed. In this review a highlight to some of the most important markers was made. Further determination of recurrence and progression marker will contribute to establish better treatments for the individual patient. Molecular staging of urological tumors will allow selecting cases that will require systemic treatment. It is necessary and important to integrate under the same objectives basic and clinical research.

Topics: Antifreeze Proteins, Type I; Antigens, Neoplasm; Biomarkers, Tumor; Cysteine Proteinase Inhibitors; Hematuria; Humans; Inhibitor of Apoptosis Proteins; Keratin-19; Keratin-20; Keratins; Microtubule-Associated Proteins; Nuclear Proteins; Predictive Value of Tests; Receptor, ErbB-2; Receptor, Fibroblast Growth Factor, Type 3; Receptors, Urokinase Plasminogen Activator; Sensitivity and Specificity; Survivin; Tissue Polypeptide Antigen; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

Paraganglioma of the urinary bladder is a rare pathologic entity with no definitive histological, immunohistochemical or molecular features to determine its malignant potency. Malignancy is essentially determined by the presence of deep local invasion, invasion of adjacent structures, and lymph node or distant metastases. So far, up to 180 cases of paraganglioma have been reported, with less than 30 being malignant. We present a case of malignant paraganglioma of the urinary bladder in a 44-year-old woman. The patient's symptoms were painless hematuria and micturitional headache. The tumor presented the characteristic "zellballen" pattern of growth and immunohistochemically was positive for all neuroendocrine markers. The patient underwent partial cystectomy and the following two postoperative years were uneventful. The literature on paraganglioma of the urinary bladder, analyzing the histological, immunohistochemical and molecular characteristics, is reviewed.

Topics: Adult; Biomarkers, Tumor; Female; Humans; Immunohistochemistry; Keratins; Paraganglioma; Treatment Outcome; Urinary Bladder; Urinary Bladder Neoplasms

Bladder cancer biomarker development has advanced significantly over the last decade, but has not yet been able to make a significant impact in the diagnosis and management of the disease. Many available markers are suitable, but do not meet the expectations of physicians and patients. Patients do not want to compromise accuracy in diagnosing bladder cancer for less-invasive tests. The review highlights the latest developments in bladder cancer biomarkers, including markers developed over the last year, and comments on the high standards placed on these markers which have delayed their widespread implementation into the urologic field.. New markers described in the last year include soluble Fas, urothelial carcinoma-associated 1 and human chorionic gonadotropin beta type II genes. The latter two markers represent the contribution of genomic technology to this field. Also described are updates to known markers, including long-term follow-up of hematuria screening, recent studies in DNA methylation for bladder cancer diagnosis and patient perspectives on bladder tumor markers.. Biomarkers for bladder cancer have been intensively scrutinized over the last decade, but despite new findings and good performance characteristics, they are currently not accepted in clinical practice.

Topics: Antigens, Neoplasm; beta-N-Acetylhexosaminidases; Biomarkers, Tumor; Carcinoembryonic Antigen; Complement Factor H; DNA Methylation; DNA Mutational Analysis; DNA, Neoplasm; Enzyme-Linked Immunosorbent Assay; Fas Ligand Protein; Gene Expression Regulation, Neoplastic; Genomics; Hematuria; Histone Acetyltransferases; Humans; Hyaluronic Acid; Hyaluronoglucosaminidase; In Situ Hybridization, Fluorescence; Inhibitor of Apoptosis Proteins; Keratins; Microsatellite Repeats; Microtubule-Associated Proteins; Molecular Diagnostic Techniques; Neoplasm Proteins; Nuclear Proteins; Patient Selection; Predictive Value of Tests; Prognosis; Proteomics; Reagent Kits, Diagnostic; Survivin; Telomerase; Transcription Factors; Urinary Bladder 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

Inflammatory myofibroblastic tumours (IMFT) may arise at any anatomical site, including lung, soft tissues, retroperitoneum and bladder. Although morphologically similar, these lesions encompass a spectrum of entities with differing aetiology, ranging from reactive/regenerative proliferations to low-grade neoplasms with a risk of local recurrence, but no significant metastatic potential. Vesical IMFT usually presents as a polypoid mass with a pale firm cut surface and can be of considerable size, mimicking a malignant tumour clinically and radiologically. Its good outcome, however, warrants conservative surgical excision, emphasising the importance of identification and distinction from malignant tumours of the bladder that may require more radical surgery and/or adjuvant therapy. We conducted a preliminary retrospective, comparative immunocytochemical study of 20 bladder tumours, including nine IMFTs, five spindle cell (sarcomatoid) carcinomas, two rhabdomyosarcomas, two leiomyosarcomas and two neurofibromas. The results confirmed IMFT positivity for smooth muscle actin, desmin and cytokeratin in 78-89% cases, resulting in potential confusion with sarcomatoid carcinoma or leiomyosarcoma. In contrast, cytoplasmic anaplastic lymphoma kinase (ALK 1) staining was present in eight IMFT (89%), but was not seen in any other lesion examined. The ALK 1 staining was confirmed by fluorescence in situ hybridisation, with translocation of the ALK gene present in 15-60% tumour cells in four of six IMFT examined, but not in four cases of sarcomatoid carcinoma or three of leiomyosarcoma. In conclusion, ALK 1 staining may be of value in the distinction of vesical IMFT from morphologically similar entities, and often reflects ALK gene translocations in these lesions.

Topics: Actins; Adolescent; Adult; Anaplastic Lymphoma Kinase; Calcium-Binding Proteins; Calmodulin-Binding Proteins; Calponins; Desmin; Diagnosis, Differential; Female; Gene Rearrangement; Granuloma, Plasma Cell; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratins; Male; Microfilament Proteins; Middle Aged; Muscle, Smooth; Protein-Tyrosine Kinases; Receptor Protein-Tyrosine Kinases; Staining and Labeling; Urinary Bladder Diseases; Urinary Bladder Neoplasms; Vimentin

An international consultation on the diagnosis of non-invasive urothelial neoplasms was held in Ancona, Italy in May 2001. Besides histology and problems of classification, one group of experts (Committee no. 3) discussed the molecular pathology and cytometry of non-invasive urothelial carcinomas. In the following first part, special immunohistochemical and molecular markers for stratifications in bladder cancer were discussed including different cytokeratins (clone 34betaE12, CK 20), cell proliferation markers (Ki67/MIB-1, PCNA, AgNOR, DNA-cytometry), tumor suppressor genes and oncogenes (p53, p21, erb-B2, bcl-2), different receptor expressions of epidermal growth factor and vascular endothelial growth factor and others. These molecular markers were analyzed in diagnosis of urothelial carcinomas, recurrences, progression and response to treatment.

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Cell Division; DNA, Neoplasm; Genes, Tumor Suppressor; Humans; Immunohistochemistry; Keratins; Molecular Biology; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Prognosis; Urinary Bladder Neoplasms

The Adenocarcinoma of the Urachus is very rare tumor, with an incidence of 1/5,000,000 inhabitants, represents less than 0.001 of all types of bladder cancer.. A 51 year old man with a chronic history of suprapubic pain and hematuria. Physical examination and excretory urography were normal. The cystoscopy demonstrated a oedematosa area in cupola of bladder wall. The transuretral biopsy was moderately differentiated adenocarcinoma, with positive antibody to CK7 and CK20, the carcinoembryonic antigen was 6.6 ng/ml. Extended partial cystectomy was done, followed for chemotherapy and radiotherapy.. The treatment of adenocarcinoma of the urachus with a combination of extended partial cystectomy, chemotherapy and radiation, is a effective treatment.

Topics: Adenocarcinoma, Mucinous; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoembryonic Antigen; Chemotherapy, Adjuvant; Cisplatin; Combined Modality Therapy; Cystectomy; Deoxycytidine; Gemcitabine; Humans; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Middle Aged; Neoplasm Proteins; Radiotherapy, Adjuvant; Urachus; Urinary Bladder Neoplasms

A number of urine based markers have been and are being investigated for the diagnosis and prognostication of urological conditions. A majority of these markers have been evaluated in urological neoplasms, particularly bladder cancer. The diagnosis of bladder cancer currently relies on identifying malignant cells in the urine and subsequently visualizing the tumor on cystoscopy. This diagnosis is further confirmed by transurethral resection or biopsy. While urine cytology is specific, it is not sensitive, especially for detecting low grade disease. This characteristic has prompted the search for more accurate markers of bladder cancer. In this review we critically examine the results of studies evaluating various markers for bladder cancer.. The published literature on urine based markers for all urological diseases, particularly bladder cancer, was identified using a MEDLINE search and critically analyzed. The sensitivity, specificity, positive and negative predictive values of the various markers were compared. The benefit of using combined markers rather than a single marker was also analyzed from published reports.. Most published literature on urine based markers for urological malignancies involve such markers for diagnosing and prognosticating bladder cancer. Hence, we focused mainly on urine based markers in bladder cancer. Most markers appear to have an advantage over urine cytology in terms of sensitivity, especially for detecting low grade, superficial tumors. However, most markers tend to be less specific than cytology, yielding more false-positive results. This scenario is more common in patients with concurrent bladder inflammation or other benign bladder conditions. However, there is reason to be optimistic about several new markers that appear to provide better specificity. Few urine based markers have been identified and investigated in other urological tumors.. Detecting bladder cancer using diagnostic markers still presents a challenge. A number of new markers are currently available that appear to be significantly more accurate than cytology. However, further studies involving a larger number of patients are required to determine their accuracy and widespread applicability for diagnosing bladder cancer. Urine based markers do not appear to have a significant role in the diagnosis or prognosis of other urological malignancies, such as prostate, kidney or testicular cancer.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Fibrin Fibrinogen Degradation Products; Flow Cytometry; Humans; Hyaluronic Acid; Hyaluronoglucosaminidase; Keratins; Nuclear Proteins; Prognosis; Sensitivity and Specificity; Telomerase; Urinary Bladder Neoplasms

The current system used to classify bladder carcinoma by stage and histological grade is very useful, yet still has limited ability to predict the natural history, or treated natural history, of a bladder tumor. Cystoscopy and urine cytology are currently the gold standard in the diagnosis and follow-up of bladder cancer. Classical urine cytology, however, at least in the diagnosis of G1 tumors, is definitely characterized by a relative low sensitivity. The low sensitivity and subjective interpretation of cytology led to the development of several tests to detect bladder cancer in urine. We provide a current, comprehensive review of the literature on bladder tumor markers and summarize their diagnostic potential. In conclusion, under the premise that cystoscopy has never been subjected to evaluation, no diagnostic marker with a sensitivity and specificity comparable to cystoscopy currently exists. The combined analysis of several tumor markers, as in the Immunocyt test, seems to be the most promising approach. In the future, rather highly sensitive tests may be able to replace cystoscopy or prolong the intervals of cystoscopies in the follow-up of selected patients.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Cystoscopy; Enzyme-Linked Immunosorbent Assay; Fibrin Fibrinogen Degradation Products; Humans; Immunoassay; Keratins; Nuclear Proteins; Sensitivity and Specificity; Telomerase; Urinalysis; Urinary Bladder Neoplasms; Urine

We describe five patients who recently presented with gross hematuria secondary to inflammatory pseudotumors of the bladder along with a review of the literature. At presentation, four of the five patients were clinically misdiagnosed as malignancies of which two were further believed to be leiomyosarcomas on initial histological examination because of their spindle-cell appearance. Conservative excision either by transurethral resection or partial cystectomy was curative in all cases. The main importance of these rare, benign lesions is to differentiate them from malignant tumors for which they may be mistaken, thus avoiding radical surgery and its attendant complications.

Topics: Adolescent; Adult; Cystectomy; Desmin; Diagnosis, Differential; Diagnostic Errors; Female; Granuloma, Plasma Cell; Hematuria; Humans; Keratins; Leiomyosarcoma; Male; Middle Aged; Urinary Bladder Diseases; Urinary Bladder Neoplasms; Vimentin

Three cases of neuroendocrine carcinoma showing skeletal muscle differentiation are presented. The tumors were located in the skin and subcutaneous tissue, the urinary bladder, and the nasal cavity respectively, and were composed by two cell types admixed intimately with each other. One cell type had features identical to those seen in conventional small cell neuroendocrine carcinoma, including scanty cytoplasm, round nuclei with fine granular chromatin, immunohistochemical reactivity for neuron-specific enolase, chromogranin and cytokeratins, and electron-dense granules on ultrastructural examination. The second cell type was either plasmacytoid or elongated and straplike, with abundant eosinophilic cytoplasm and irregular nuclei with prominent nucleoli. These cells showed immunohistochemical positivity for desmin, sarcomeric actin, myoglobin, and myogenin. They also exhibited ultrastructural evidence of rhabdomyoblastic differentiation in the form of contractile filaments with abortive Z-band formation. An origin from a cell capable of dual differentiation toward neuroendocrine and rhabdomyoblastic elements is postulated for these tumors.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Neuroendocrine; Chromogranins; Cytoplasmic Granules; Fatal Outcome; Female; Humans; Immunoenzyme Techniques; Keratins; Male; Muscle, Skeletal; Nose Neoplasms; Phosphopyruvate Hydratase; Rhabdomyosarcoma; Skin Neoplasms; Urinary Bladder Neoplasms

The cytokeratins are the intermediate filament proteins characteristic of epithelial cells. In human cells, some 20 different cytokeratin isotypes have been identified. Epithelial cells express between two and ten cytokeratin isotypes and the consequent profile which reflects both epithelial type and differentiation status may be useful in tumour diagnosis. The transitional epithelium or urothelium of the urinary tract shows alterations in the expression and configuration of cytokeratin isotypes related to stratification and differentiation. In transitional cell carcinoma, changes in cytokeratin profile may provide information of potential diagnostic and prognostic significance. The intensification of immunolabelling with some CK8 and CK18 antibodies may underly an active role in tumour invasion and foci of CK17-positive cells may represent proliferating populations. Loss of CK13 is a marker of grade and stage and de novo expression of CK14 is indicative of squamous differentiation and an unfavourable prognosis. However, perhaps the most important recent finding is the demonstration that a normal CK20 expression pattern is predictive of tumour non-recurrence and can be used to make an objective differential diagnosis between transitional cell papilloma and carcinoma. This review will consider cytokeratin expression in urothelium and discuss the application of cytokeratin typing to the diagnosis and prognosis of patients with TCC.

Topics: Biomarkers; Carcinoma, Transitional Cell; Cell Differentiation; Humans; Intermediate Filaments; Keratins; Protein Isoforms; Urinary Bladder Neoplasms; Urothelium

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

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

Herein is a review of clear cell neoplasms of selected sites in the urinary tract and male reproductive system, including the kidney, the urinary bladder, testis, epididymis, and prostate. Clear cell cytoplasmic alteration in neoplasms at these sites is a relatively common light microscopic finding. Examples of such neoplasms with clear cell change include the clear cell type of renal cell carcinoma, clear cell adenocarcinoma of urethra and bladder, the classic type of seminoma, papillary cystadenoma of the epididymis, and well-differentiated adenocarcinoma of the prostate. Of importance, numerous non-neoplastic benign entities may also manifest cleared cytoplasm and therefore are presented in the differential in this review. Indeed, knowledge of the neoplastic and non-neoplastic entities displaying clear cell change at each anatomic site should enable the surgical pathologist to approach the differential diagnosis of these conditions in a more logical and rigorous fashion.

Topics: Adenocarcinoma; Alkaline Phosphatase; Biomarkers, Tumor; Carcinoma, Renal Cell; Diagnosis, Differential; Genital Neoplasms, Male; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Male; Mucin-1; Prostatic Neoplasms; Testicular Neoplasms; Urinary Bladder Neoplasms; Urologic Neoplasms

A tumor mass resected from the anterior bladder wall of a 68-year-old woman displayed unusual histologic features: sheets of hepatoid cells merging focally with a secondary glandular pattern of adenocarcinoma. Intracytoplasmic hyaline globules and bile production within the solid areas supported the impression of hepatocytic differentiation. Immunoreactivity for alpha-fetoprotein (AFP) and alpha-1-antitrypsin and a striking canalicular immunostaining pattern for carcinoembryonic antigen and epithelial membrane antigen all indicate hepatocellular differentiation within this bladder tumor. This represents a case of a hepatoid adenocarcinoma located in the urinary bladder. The use of the term "hepatoid" in the literature is reviewed and the reported cases are grouped into two distinct categories of tumors: (1) germ cell tumors with focal hepatoid areas and (2) true hepatoid adenocarcinomas that meet histologic and immunohistochemical criteria for hepatocellular differentiation. AFP-producing tumors without any other feature of hepatocellular differentiation should not be considered as hepatoid tumors. This classification of hepatoid tumors is likely to be important in elucidating the histogenesis and clinicopathologic features of these unusual neoplasms.

Topics: Adenocarcinoma; Aged; alpha 1-Antitrypsin; alpha-Fetoproteins; Antigens, Neoplasm; Carcinoembryonic Antigen; Carcinoma, Endometrioid; Carcinoma, Transitional Cell; Cell Nucleus; Cytoplasm; Female; Humans; Keratins; Liver; Membrane Glycoproteins; Mucin-1; Ureteral Neoplasms; Urinary Bladder Neoplasms; Uterine Neoplasms

The diagnosis and classification of bladder cancer are based primarily on histologic and cytologic light microscopy. The significant cytologic alternations are abnormal (increased) DNA content and structural changes in chromatin. Measurements of DNA content carried out by flow cytometry on suspensions of tumor cells and bladder irrigation specimens correlate well with clinical and biopsy findings. Badalament et al (Badalament RA, Kimmel M, Gay H, et al. Cancer 1987;59:2078-2085) reported that a single bladder wash flow cytometry correctly detected 83% of bladder cancers. The technique is most sensitive in detecting early, in situ, and superficially invasive carcinoma. In 100 urologic patients with non-neoplastic disease of the bladder, Klein et al (Klein FA, Herr HW, Sogani PC, et al. J Urol. 1982;127:946-948) reported only two false-positive examinations. Flow cytometry appears to be at least as valuable as conventional urinary cytology, without the need of an experienced cytopathologist. However, as specimens must be collected by bladder irrigation via catheter or cytoscope, the technique is most suitable not for population screening, but in monitoring high-risk populations: industrial workers or others exposed to carcinogens, persons with a history of urothelial tumors, and adult patients referred for urologic examinations because of hematuria, unexplained, recurrent cystitis, or other urologic symptoms. DNA flow cytometry also has been valuable in monitoring the conservative treatment of bladder cancer and of predictive value in the intravesical bacille Calmette-Guérin treatment of superficial carcinomas of the bladder. Dual parameter measurements of DNA content and antigen expression are now under evaluation, as are measurements of chromatin structure alteration, metabolism, and proliferative rate. These promise to discriminate subsets of bladder cancer that may be predictive of clinical behavior.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Blood Group Antigens; Carcinoma, Papillary; DNA, Neoplasm; Flow Cytometry; Humans; Keratins; Urinary Bladder Neoplasms

Immunohistochemistry of intermediate filaments (IF) is a new and important way to evaluate the epithelial, mesenchymal, muscular, glial, or neural differentiation in tumors. This is based on the stable cell-type-specific expression of IF proteins in normal and neoplastic tissues. Immunohistochemical studies with antibodies to intermediate filaments have also given new perspectives in the histogenesis and biologic nature of many tumors. This article reviews both the recent findings and the authors' experience in the use of intermediate filament antibodies in tumor diagnosis and classification.

Topics: Adenocarcinoma; Antibodies, Neoplasm; Carcinoma, Squamous Cell; Desmin; Diagnosis, Differential; Glial Fibrillary Acidic Protein; Histocytochemistry; Humans; Immunochemistry; Keratins; Melanoma; Mesothelioma; Microfilament Proteins; Microscopy, Electron; Neoplasm Proteins; Neoplasms; Nervous System Neoplasms; Sarcoma; Soft Tissue Neoplasms; Thyroid Neoplasms; Urinary Bladder Neoplasms; Vimentin

Topics: Animals; Basement Membrane; Carcinogens; Cell Membrane; Cyclophosphamide; Epithelium; Female; Glycoproteins; Golgi Apparatus; Humans; Keratins; Male; Microscopy, Electron; Neoplasm Proteins; Neoplasms, Experimental; Polysaccharides; Urinary Bladder; Urinary Bladder Neoplasms

The aim of this study was to determine the diagnostic accuracy of UBC. This prospective phase II study was conducted at four Swedish hospitals. UBC Rapid was evaluated in four groups: A, newly diagnosed bladder cancer (n = 94); B, follow-up of non-muscle-invasive bladder cancer (n = 75); C, benign urinary tract diseases (n = 51); and D, healthy controls (n = 50). Tumours were divided into high risk (carcinoma in situ, TaG3, T1, T2 and T3) and low risk (low malignant potential, TaG1 and TaG2). Urine samples were quantitatively analysed by UBC Rapid. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated based on optimal cut-off (receiver operator characteristics curve analysis). A linear regression compared the UBC Rapid results in the different risk groups.. The optimal cut-off was 8.1 μg/l. The median UBC Rapid values were 9.3 μg/l [interquartile range (IQR) 30.9] and 4.3 μg/l (IQR 7.8) in patients with positive and negative cystoscopy, respectively (p < .001). The value for group A was 15.6 μg/l (IQR 37.9), group B 5.6 μg/l (IQR 8.6), group C 5.1 μg/l (IQR 9.0) and group D 3.3 μg/l (IQR 7.1). Sensitivity was 70.8%, specificity 61.4%, PPV 71.3% and NPV 60.8%. The high-risk group had significantly higher UBC Rapid values than the low-risk group: 20.5 μg/l (IQR 42.2), sensitivity 79.2% and specificity 61.4% versus 7.0 μg/l (IQR 9.9), sensitivity 60.0% and specificity 61.4% (p = .039).. The UBC Rapid urine-based marker for bladder cancer gave higher values in patients with positive than in those with negative cystoscopy. The diagnostic accuracy was better in patients with high-risk than in those with low-risk tumours, and was better during primary detection than during surveillance.

Topics: Aged; Aged, 80 and over; Female; Humans; Keratins; Male; Middle Aged; Neoplasm Grading; Neoplasm Invasiveness; Neoplasm Staging; Photometry; Point-of-Care Systems; Prospective Studies; Sensitivity and Specificity; Sweden; Urinary Bladder Diseases; Urinary Bladder Neoplasms

To assess the value of urine tumor-associated trypsin inhibitor (TATI), CYFRA 21-1, which measures cytokeratin 19 fragment, and urinary bladder carcinoma antigen (UBC) for the detection of high-grade bladder carcinoma.. A total of 160 individuals were enrolled in the present study. Of these, 80 were patients with proven primary high-grade urothelial bladder cancer (group 1), 40 were healthy volunteers (group 2), and 40 had history of benign urologic disease (group 3). All were evaluated with respect to urinary TATI, CYFRA 21-1, and UBC levels. All these markers were evaluated using commercial kits. Cytology was also performed.. The TATI measurements were significant greater in group 1 compared with groups 2 and 3. The cutoff point used for TATI, CYFRA 21-1, and UBC was 22, 2.8, and 12 microg/L, respectively. The overall sensitivity was 85.7% for TATI, 61.9% for CYFRA 21-1, 50% for UBC, and 42.8% for cytology. TATI was significantly more sensitive in Stage Ta (80%) than was CYFRA 21-1 (32%), UBC (12%), and cytology (20%). TATI was also more sensitive compared with other tumor markers for Stage T1 but not for Stage T2 or T3.. The results of our study have shown that TATI is a promising urinary tumor marker for high-grade urothelial bladder cancer. It is more sensitive than CYFRA 21-1, UBC, and cytology for Stage Ta and T1 bladder cancer.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Humans; Keratin-19; Keratins; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Trypsin Inhibitor, Kazal Pancreatic; Urinary Bladder Neoplasms

The objective of this study was to clarify the significance of micrometastases in pelvic lymph nodes in patients who underwent radical cystectomy for bladder cancer.. We included 40 patients with locally invasive bladder cancer who underwent radical cystectomy and pelvic lymphadenectomy. Expression of cytokeratin 19 (CK19), uroplakin II (UP II), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in 760 lymph nodes were assessed by a fully quantitative real-time reverse transcription-PCR (RT-PCR) assay. The quantification value of CK19 or UP II mRNA was described as each value relative to GAPDH mRNA. In this study, we regarded specimen in which either CK19 or UP II mRNA was positive as "presence of micrometastasis.". Routine pathologic examinations detected tumor cells in 29 lymph nodes from six patients. Real-time RT-PCR identified positive expression of CK19 and UP II mRNAs in 49 lymph nodes from 10 patients and 98 lymph nodes from 16 patients, respectively. Of 633 lymph nodes from 34 patients with no pathologic evidence of nodal involvement, 13 nodes from five patients and 58 nodes from 10 patients were diagnosed as positive for CK19 and UP II mRNAs expression, respectively, by real-time RT-PCR. Presence of micrometastases was significantly associated with other conventional prognostic variables, including pathologic stage and microvascular invasion. Disease recurrence was occurred in eight patients, among whom four patients were negative for lymph node metastasis by routine pathologic examination and diagnosed as having micrometastasis by real-time RT-PCR assay. Furthermore, cause-specific survival rate in patients without micrometastasis was significantly higher than that in those with micrometastasis, irrespective of the presence of pathologic-positive nodes.. Approximately 30% of locally invasive bladder cancer shed cancer cells to pelvic lymph nodes, and disease recurrence after radical cystectomy could be explained, at least in part, by micrometastases in pelvic lymph nodes.

Topics: Cystectomy; Female; Gene Expression Profiling; Humans; Keratins; Lymphatic Metastasis; Male; Membrane Proteins; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Pelvis; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; Urinary Bladder Neoplasms; Uroplakin II

Existing clinical data have shown that high-grade prostatic intraepithelial neoplasia (HGPIN) is the most likely precursor to prostate cancer (CaP). Criteria to distinguish HGPIN that progress to CaP from those that do not remain poorly defined. Our objective was to evaluate microvessel density as a molecular marker for distinguishing HGPINs that have the potential of progressing to cancer.. Human prostatic tissue samples were collected randomly from 50 prostatectomy and cystoprostatectomy patients. Formalin-fixed and paraffin-embedded sections were used for immunohistochemical localization of rabbit anti-human von Willebrand factor VIII (vWF) IgG, mouse anti-high molecular weight cytokeratin 34BE-12 in basal cells, and mouse anti-heparan sulphate proteoglycan (HSPG) IgGs in basement membranes associated with benign prostatic hyperplasia (BPH), PIN associated with some BPH (isolated PIN), and PIN associated with CaP.. Analysis of immunostaining data showed that PINs could be categorized according to their distributions within and outside 2 standard deviations (SD) of the mean for microvessel density. The average number of microvessels was significantly higher (P < 0.0001) in PINs associated with Gleason score 7 tumors than those associated with Gleason scores 4-6 (P < 0.1328) or 8 and 9 tumors (P < 0.1708). Morphologically, PINs within 2 SD were composed of low- and high-grade type, whereas those outside 2 SD of microvessel density were predominantly of high-grade type. Cytokeratin and HSPG localization patterns also showed differences in PINs found within and outside 2 SD of microvessel density. We found localization of cytokeratin 34BE-12 in basal cells of specimens with BPH alone, isolated PIN, and PIN associated with CaP within 2 SD, whereas many PINs outside 2 SD showed disruptions in cytokeratin localization. The basement membranes of PINs within 2 SD of microvessel density were relatively intact, whereas those outside 2 SD were fragmented.. Our immunostaining data indicates that once HGPIN is found in the initial prostatic biopsy, it should be evaluated for microvessel density by localization of vWF. Our data indicate that characteristics of HGPIN can be augmented by evaluations of cytokeratin and HSPG molecular markers to assess the potential of HGPIN progression to malignancy. When biopsy samples show HGPIN with increased microvessel density and disrupted cytokeratin and HSPG markers, the patient may be a candidate for repeat biopsy. Since our study is limited to 50 prostate tissue samples, we emphasize that our conclusion is tentative and ought to be confirmed in a study with a larger sample size. This is the first report to show that microvessel density may distinguish HGPIN that is a precursor to prostate cancer.

Topics: Aged; Biomarkers, Tumor; Disease Progression; Heparan Sulfate Proteoglycans; Humans; Immunoenzyme Techniques; Keratins; Male; Microcirculation; Middle Aged; Neovascularization, Pathologic; Prostatectomy; Prostatic Hyperplasia; Prostatic Intraepithelial Neoplasia; Prostatic Neoplasms; Urinary Bladder Neoplasms; von Willebrand Factor

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

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

We evaluated CYFRA 21-1, an immunoradiometric assay, developed to detect soluble cytokaratin 19 fragment, for its diagnostic performance in bladder transitional cell carcinoma as well as its analytical performance.. We assessed CYFRA 21-1 in the serum and urine of 182 patients, including 66 with bladder transitional cell carcinoma (group 1), 66 with another urological pathology (group 2) and 50 free of urothelial disease (group 3). The power of urinary CYFRA as a diagnostic test for bladder transitional cell carcinoma was evaluated by receiver operating characteristics curve analysis. Analytical performance was assessed by determining intra-assay and interassay precision, and accuracy by dilution testing and recovery of supplemented analyte.. Mean urinary CYFRA plus or minus standard deviation was 154.39+/-49.00, 22.6+/-8.9 and 2.40+/-0.14 ng./ml. in groups 1 to 3, respectively (significantly different). Receiver operating characteristics curve analysis revealed optimal 96.9% sensitivity and 67.2% specificity for a threshold value of 4 ng./ml. Analytical determination showed that intra-assay and interassay precision provides a satisfactory coefficient of variation. The assay for accuracy had acceptable recovery in diluted samples as well as in those with supplemented analyte.. The urinary immunoradiometric CYFRA 21-1 assay performs well analytically. Urinary CYFRA 21-1 is a useful marker for diagnosing transitional cell carcinoma and provides sensitivity in low grade disease.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Female; Humans; Keratin-19; Keratins; Male; Middle Aged; Prospective Studies; Reproducibility of Results; Urinary Bladder Neoplasms

Our objectives were to evaluate the diagnostic value of two new urinary tumor markers, cytokeratin 18 (CK18) and bladder tumor fibronectin (BTF), for the detection and monitoring of bladder cancer. The study comprised 931 urine samples belonging to 402 subjects: 112 individuals under suspicion for a primary bladder tumor (group 1); 104 bladder cancer patients under scheduled follow-up (group 2); 109 bladder cancer patients receiving intravesical instillations (group 3); 45 patients with other urological diseases (group 4); and 32 healthy subjects (group 5). Voided urine samples were collected before cystoscopies, between them and before intravesical instillations. CK18 and BTF tests were measured by chemiluminescent immunoassays. Optimal receiver operating characteristic cutoffs of 7.4 microg/L for CK18 and 52.8 microg/liter for BTF rendered overall sensitivities of 66.2% for CK18 and 80.0% for BTF at specificities of 88.4 and 74.7%, respectively. Urinary cytology provided a sensitivity of 29.2% at a specificity of 99.1%. Sensitivities were 80.8, 74.2, and 82.3% for BTF and 71.1, 77.4, and 64.7% for CK18 for groups 1 to 3, respectively. False positive rates were higher for BTF in all groups of patients. Elevated urinary tumor markers during the monitoring of patients with bladder cancer could detect recurrence sooner than scheduled cystoscopies. Persistence of negative markers was greatly indicative of free of disease status in follow-up. CK18 and BTF in urine may eventually prove to be of benefit for specific patients with bladder carcinoma given its higher sensitivity compared with cytology. In selected patients, namely those with persistent negative urinary CK18 and BTF, the number of cystoscopies could be reduced.

Topics: Adult; Aged; Aged, 80 and over; Antibodies; Biomarkers, Tumor; Cystoscopy; False Positive Reactions; Female; Fibronectins; Humans; Keratins; Male; Middle Aged; Monitoring, Physiologic; Peptide Fragments; Protein Structure, Tertiary; ROC Curve; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urologic Diseases

To study the diagnostic performance of the Urinary Bladder Cancer (UBC) test in patients with superficial bladder carcinoma.. One hundred one patients in follow-up for superficial bladder cancer (pTa, pT1, carcinoma in situ) were recruited for this study. Each patient underwent cystoscopy and transurethral resection or biopsy, with subsequent histologic confirmation in the case of abnormalities. In addition, specimens were assessed with an immunoenzymometric assay for cytokeratin expression (the UBC test), and the urinary creatinine concentration was determined to correct for different degrees of urinary dilution. Different methods were applied to calculate the diagnostic value of the UBC test.. Both noncorrected and corrected median values of the UBC test were comparable between patients with and without a recurrent bladder tumor. The overall sensitivity, specificity, and positive and negative predictive values of the noncorrected UBC test was 20.7%, 84.7%, 35.3%, and 72.6%, respectively. For the corrected UBC test, the corresponding values were 20.7%, 79.2%, 28.6%, and 71.3%. The area under the receiver operating characteristic curve was not significantly different from 0.50, indicating no diagnostic value of the UBC test in this study.. The diagnostic value of this new urinary marker appears insufficient for the follow-up of patients with superficial bladder cancer.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy; Carcinoma in Situ; Creatinine; Cystoscopy; Female; Follow-Up Studies; Humans; Keratins; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Predictive Value of Tests; Regression Analysis; ROC Curve; Sensitivity and Specificity; Statistics, Nonparametric; Urinary Bladder Neoplasms

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

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

We evaluated the diagnostic performance of the new noninvasive bladder cancer test on voided urine samples from patients with transitional cell carcinoma compared to symptomatic and asymptomatic controls.. Urinary bladder cancer antigen was measured in urine from 86 patients with active transitional cell carcinoma of the bladder (group 1), 76 patients free of transitional cell carcinoma as confirmed by cystoscopy at followup (group 2), 25 patients with other benign urological diseases (group 3), 25 patients with other malignant pathological conditions (group 4) and 30 healthy subjects free of urological diseases (group 5).. Mean urinary bladder cancer antigen concentrations were 104.84, 4.57, 11.79, 48.87 and 1.38 microg/l, for groups 1 to 5, respectively, which was statistically different (p = 0.00005) except for groups 1 and 4 (p = 0.187). Sensitivity was 87.0% (95% confidence interval 79.2 to 92.7) and specificity was 86.8% (77.1 to 93.5%), and both were optimized by receiver operating characteristics plot analysis at a threshold value of 9.74 microg/l using asymptomatic (group 2) compared to known cancer (group 1) cases.. Urinary bladder cancer antigen might have a role as a potential tumor marker for diagnosing transitional cell carcinoma of the bladder.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Female; Humans; Keratins; Male; Middle Aged; ROC Curve; Sensitivity and Specificity; Urinary Bladder Neoplasms

A new electrochemiluminescent immunoassay (ECLIA) has been developed for the determination of cytokeratin 19 (CYFRA 21-1) in the Elecsys 2010 immunoassay system. Urinary CYFRA 21-1 might have a role in the diagnosis of bladder cancer.. We performed an analytical evaluation of the CYFRA 21-1 ECLIA for serum and urine samples. The clinical value of urinary CYFRA 21-1 for the detection of bladder cancer was evaluated through its measurement in 226 urine samples from symptomatic and asymptomatic controls.. At concentrations of 2-30 microg/L, within-assay imprecision (CV) was below 2.1% for sera and 3.3% for urines, with interassay CVs below 3.3% for sera and 4.9% for urines. The day-to-day CV was <20% at concentrations >0.2 microg/L (functional sensitivity). Measurement of diluted samples showed that the assay estimated CYFRA 21-1 between 98% and 103% for sera and 98% and 105% for urines. Recovery of added CYFRA 21-1 was 99-105% for sera and 96-115% for urines. We separately compared serum and urine CYFRA 21-1 ECLIA results with those obtained with an IRMA (CIS bio international). Regression analysis for sera was: CYFRA 21-1 (ECLIA) = 0.520 + 1.018 CYFRA 21-1 (IRMA); [95% confidence interval (CI) (y-intercept), -0.260 to 1.309]; 95% CI (slope), 0.978-1.060; n = 100; S(y|x) = 3.242; r(2) = 0.987. For urine samples it was: CYFRA 21-1 (ECLIA) = 0.716 + 0.966 CYFRA 21-1 (IRMA); 95% CI (y-intercept), 0.009-1.422; 95% CI (slope), 0.956-0.976; n = 100; S(y|x) = 4.136; r(2) = 0.986. In urine samples voided by patients with and without bladder cancer, the best ROC analysis discrimination provided 81.0% (95% CI, 72.7-87.7%) sensitivity and 97.2% (95% CI, 90.2-99.6%) specificity at a threshold value of 5.7 microg/L.. Our initial evaluation showed reliable analytical performance for urinary CYFRA 21-1, which might assist urologists in the detection of bladder cancer as a noninvasive adjunct to cystoscopy.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Electricity; Female; Humans; Immunoassay; Keratin-19; Keratins; Luminescent Measurements; Male; Middle Aged; Reproducibility of Results; ROC Curve; Urinary Bladder Neoplasms

CYFRA 21-1, an immunoradiometric assay developed for the detection of a soluble cytokeratin 19 fragment, is evaluated for its diagnostic performance in urine of patients with transitional cell carcinoma.. CYFRA 21-1 was investigated in serum and urine of 128 patients, including 48 with bladder transitional cell carcinoma (group 1), 44 with other urological pathological conditions (group 2) and 36 free of urothelial disease (group 3). Urinary cytopathology was also performed.. Mean urinary CYFRA was 123.5 +/- 53, 11.9 +/- 4.8 and 2.3 +/- 0.2 ng./ml. for groups 1 to 3, respectively, and was significantly different. From the receiver operating characteristics curve, the optimal combination of 96% sensitivity and 74% specificity was determined for a threshold value of 4 ng./ml. while overall cytopathology sensitivity was 43%.. Urinary CYFRA 21-1 may be a useful marker for diagnosing transitional cell carcinoma.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Transitional Cell; Female; Humans; Immunoradiometric Assay; Keratins; Male; Middle Aged; ROC Curve; Sensitivity and Specificity; Urinary Bladder Neoplasms

CYFRA 21-1 is a new tumor marker that measures cytokeratin 19 (CK-19) fragments by a sandwich enzyme-linked immunosorbent assay (ELISA). In this study, we evaluated the usefulness of serum and urine CYFRA 21-1 as a tumor marker for bladder cancers.. We measured serum and urine CYFRA 21-1 levels in a group of patients with bladder cancer (n = 58) and a group without bladder cancer (n = 220). The latter group was divided into five subgroups of patients (those with cystitis, benign prostatic hyperplasia (BPH), urolithiasis, or renal dysfunction, and a group of healthy, controls). In the bladder cancer group, we measured CYFRA 21-1 levels after transurethral resection of bladder tumor (TUR-Bt), and we also analyzed the relationship between serum and urine CYFRA 21-1 levels and tumor-related factors.. Serum and urine CYFRA 21-1 levels were significantly higher in the bladder cancer group than in each of the non-bladder cancer subgroups. However, urine CYFRA 21-1 levels did not differ significantly between the bladder cancer group and the cystitis subgroup. In the bladder cancer group, serum CYFRA 21-1 levels were significantly higher in patients with local advanced-stage tumors and in those who had metastases. Urine CYFRA 21-1 levels decreased as a function of time after TUR-Bt. These levels were strongly correlated with tumor volume and were significantly better than urine cytology for the detection of bladder cancers of grades 1 and 2.. These results suggest that urine CYFRA 21-1 is a useful tumor marker in screening for bladder cancer, and that serum CYFRA 21-1 may be a tumor marker for advanced bladder cancers.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Enzyme-Linked Immunosorbent Assay; Humans; Keratins; ROC Curve; Urinary Bladder Neoplasms; Urine

Combined analysis of cytokeratin 7 (CK7) and cytokeratin 20 (CK20) is often used for assessing the origin of metastatic cancer. To evaluate the diagnostic utility of CK7 and CK20, tissue microarrays containing 15,424 samples from 120 different tumor types and subtypes and 608 samples of 76 different normal tissue types were analyzed by immunohistochemistry. CK7 positivity was seen in 52% (8.7% weak, 5.9% moderate, 37% strong) and CK20 positivity in 23% (5.1% weak, 3.4% moderate, 15% strong) of interpretable tumors. Of 8390 positive tumors, 1181 (14%) showed positivity for CK7 and CK20, 5380 (64%) showed positivity for CK7 alone, and 1829 (22%) showed positivity for CK20 alone. CK20 predominated in gastrointestinal tract, urothelial and Merkel cell carcinomas. CK7 was usually negative in prostate cancer and colorectal cancer. Combined evaluation of CK7/CK20 revealed the best diagnostic utility in CK20 positive tumors, where CK7 negativity is often linked to colorectal origin while CK7 positivity argues for urothelial origin or mucinous ovarian cancer. Associations with unfavorable tumor features were found for cytokeratin 7 loss in breast cancer of no special type, urothelial and renal cell carcinomas, for CK7 overexpression in high-grade serous ovarian and gastric cancer, and for CK20 overexpression in urothelial carcinoma. CK20 loss was linked to MSI in gastric (p = 0.0291) and colorectal adenocarcinoma (p < 0.0001). These analyses provide comprehensive data on the frequency of CK7 and CK20 immunostaining - alone or in combination - in human cancers. These data facilitate interpretation of CK7/CK20 immunostaining in cancers.

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Colorectal Neoplasms; Humans; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Urinary Bladder Neoplasms

Plasmacytoid urothelial carcinoma (PUC) is a rare but clinically aggressive variant of high-grade urothelial carcinoma (HGUC). Cytological features include single plasmacytoid neoplastic cells with N:C ratio around 0.5, eccentric nuclei, nuclear hyperchromasia, irregular nuclear membrane, and vacuolated cytoplasm. Micropapillary urothelial carcinoma (MPUC) is another clinically aggressive variant of HGUC that shares some overlapping features of PUC. The diagnosis of these two aggressive variants in pleural effusions can be challenging due to features mimicking adenocarcinoma, unusual immunochemistry profile, and confusion with differential diagnoses, especially when pertinent clinical information is unavailable. We present report on one case each of pleural fluid metastasis of PUC and MPUC respectively, and compare the findings with that of a metastatic conventional HGUC originally thought to be metastatic adenocarcinoma. The diagnosis of PUC was confirmed with immunohistochemical studies showing expression for cytokeratin, GATA-3, uroplakin II, and CD138, diminished or loss of E-cadherin membranous expression, negative expression for p63, p53, Epicam-BerEP4, Epicam-MOC31, and p120. The diagnosis of MPUC was confirmed with immunostain profile similar to that of PUC except positive stain for E-cadherin, p120, and p53. The diagnosis of HGUC was confirmed with immunohistochemical studies showing expression for cytokeratin, GATA-3, uroplakin II, p63, Epicam-BerEP4 (focal weak), and Epicam-MOC31. Our cases of metastatic urothelial carcinoma showed features mimicking adenocarcinoma and others, especially the MPUC and HGUC were diagnosed without prior tissue diagnosis of urothelial carcinoma. This report emphasizes the cytohistological and immunohistochemical details of urothelial carcinoma involving effusion fluid and discusses potential pitfalls in diagnosis.

Topics: Adenocarcinoma; Cadherins; Carcinoma, Papillary; Carcinoma, Transitional Cell; Humans; Keratins; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Uroplakin II; Urothelium

The genetic features of primary lymphoepithelioma-like carcinoma (LELC) of the upper urinary tract have not been systematically explored. In this study, tumor mutation profiling was performed using whole-genome sequencing in two patients with LELC of the renal pelvis. Novel candidate variants relevant to known disease genes were selected using rare-variant burden analysis. Subsequently, a population-based study was performed using the Surveillance, Epidemiology, and End Results (SEER), PubMed, MEDLINE, Embase, and Scopus databases to explore clinical features and prognostic risk factors. Immunohistochemical analysis revealed seven positive cytokeratin-associated markers in tumor cells and five positive lymphocyte-associated markers in and around the tumor area. Sub-sequently, we identified KDM6A as the susceptibility gene and LEPR as the driver gene by Sanger sequencing in case 2 of LELC of the renal pelvis. Three mutation sites of the existing targeted drugs were screened: CA9, a therapeutic target for zonisamide; ARVCF, a therapeutic target for bupropion; and PLOD3, a therapeutic target for vitamin C. In a population-based study, patients with primary LELC of the upper urinary tract had clinical outcomes similar to those of patients with primary upper urinary tract urothelial carcinoma (UUT-UC) before and after propensity score matching at 1 : 5. Focal subtype was an independent prognostic factor for the overall survival of patients with LELC of the upper urinary tract. The carcinogenesis of primary LELC may be due to different genetic variations, including single-nucleotide variants, insertion and deletions, structural variations, and repeat regions, which may provide the basis for clinical diagnosis and treatment. The prognosis of LELC in the upper urinary tract is similar to that of UUT-UC. We suggest that the focal subtype can serve as a prognostic factor for LELC of the upper urinary tract; however, further studies are required to confirm this.

Topics: Ascorbic Acid; Bupropion; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Histone Demethylases; Humans; Keratins; Kidney Pelvis; Nucleotides; Prognosis; Urinary Bladder Neoplasms; Zonisamide

Expression levels of collagens have been implicated in the progression of various cancers and interaction with cytokeratins but are not well studied in bladder cancer (BC). Therefore, we analyzed the gene and protein expression levels of collagen 1A1 (Col1a1/COL1A1), collagen 3A1 (col3a1/COL3A1), collagen 5A2 (col5a2/COL5A2), cytokeratin 14 (krt14/CK14), and cytokeratin 17 (krt17/CK17) in urothelial BC samples of different stages.. In total, 102 fresh frozen and 190 formalin-fixed and paraffin-embedded (FFPE) samples were tested using immunohistochemistry and RT-qPCR. Expression levels were correlated with clinicopathological and follow-up data.. Col1a1, col3a1, col5a2 and krt14 mRNA levels were significantly overexpressed in high-grade and muscle-invasive BC (MIBC) compared to low-grade and non-muscle invasive BC (NMIBC) cases. Disease-specific survival (DSS) was shorter in patients with high expression levels of col1a1 (p = 0.004), col3a1 (p = 0.004), and col5a2 (p = 0.028). CK14 (p = 0.020), COL3A1 (p = 0.006), and Col5A2 (p = 0.006) protein expression levels were significantly higher and protein expression levels of CK17 (p = 0.05) were significantly lower in MIBC compared to NMIBC. Furthermore, CK14 (p = 0.002) and COL5A2 (p = 0.006) protein expression level were significantly higher in high-grade compared to low-grade BC. DSS was shorter in patients with high expression levels of COL5A2 (p = 0.033) and CK14 (p = 0.042).. Expression changes of collagens and cytokeratins are univariable prognostic markers in BC.

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Collagen; Humans; Keratins; Prognosis; Urinary Bladder Neoplasms

Circulating tumor cells (CTCs) in the blood of cancer patients are of high clinical relevance. Since detection and isolation of CTCs often rely on cell dimensions, knowledge of their size is key. We analyzed the median CTC size in a large cohort of breast (BC), prostate (PC), colorectal (CRC), and bladder (BLC) cancer patients. Images of patient-derived CTCs acquired on cartridges of the FDA-cleared CellSearch

Topics: Breast Neoplasms; Cell Count; Cell Line, Tumor; Cell Nucleus; Cell Size; Cohort Studies; Colonic Neoplasms; Female; Humans; Indoles; Keratins; Male; Neoplastic Cells, Circulating; Prostatic Neoplasms; Urinary Bladder Neoplasms

Several experimental models including patient biopsies, animal models, and cell lines have been recommended to study the mechanism of bladder cancer development. After several passages in culture, cell lines lose some original features, and no longer resemble the cells of their original tumor. This makes it necessary to establish various cell lines. In an attempt to establish a new cell line for bladder cancer, JAM-ICR (RRID: CVCL_A9QB) was derived from a 64-year-old man diagnosed with a high-grade tumor. This cell line was characterized in multiple experiments involving morphological studies, immunophenotyping (by immunohistochemistry and flow cytometry), karyotyping, short tandem repeat analysis, colony-forming assays, migration and invasion assays, and chemosensitivity to anti-cancer drugs. JAM-ICR cells are pale with an irregular polygonal shape, and show some similarities to mesenchymal stem cells but with a wider shape and shorter arms. Phenotypic assessment demonstrated the simultaneous expression of mesenchymal-(vimentin, desmin, CD29, CD90, and CD106) and epithelial lineage (pan-cytokeratin) markers, which supports a phenotype similar to epithelial-mesenchymal transition for this cell line. JAM-ICR displayed high metastatic potential and stem-like properties, i.e., self-renewal, colony forming, and the coexpression of TRA-1 with CD44 and CD166. Furthermore, this cell line was significantly more resistant to doxorubicin in comparison to the 5637 cell line. These features make JAM-ICR a new bladder cancer cell line with metastatic potential and stem-like properties, which may be potentially useful as a model to elucidate the molecular and cellular mechanisms of bladder cancer pathogenesis or evaluate new drugs.

Topics: Antigens, CD; Antineoplastic Agents; Cell Line, Tumor; Cell Movement; Desmin; Dicarboxylic Acids; Doxorubicin; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Epithelial-Mesenchymal Transition; Humans; Keratins; Male; Middle Aged; Neoplasm Invasiveness; Oxazines; Urinary Bladder Neoplasms; Vimentin

Environmental exposure to arsenite (As3+) has a strong association with the development of human urothelial cancer (UC) and is the 5th most common cancer in men and the 12th most common cancer in women. Muscle invasive urothelial cancer (MIUC) are grouped into basal or luminal molecular subtypes based on their gene expression profile. The basal subtype is more aggressive and can be associated with squamous differentiation, characterized by high expression of keratins (KRT1, 5, 6, 14, and 16) and epidermal growth factor receptor (EGFR) within the tumors. The luminal subtype is less aggressive and is predominately characterized by elevated gene expression of peroxisome proliferator-activated receptor- gamma (PPARγ) and forkhead box protein A1 (FOXA1). We have previously shown that As3+-transformed urothelial cells (As-T) exhibit a basal subtype of UC expressing genes associated with squamous differentiation. We hypothesized that the molecular subtype of the As-T cells could be altered by inducing the expression of PPARγ and/or inhibiting the proliferation of the cells. Non-transformed and As-T cells were treated with Troglitazone (TG, PPARG agonist, 10 μM), PD153035 (PD, an EGFR inhibitor, 1 μM) or a combination of TG and PD for 3 days. The results obtained demonstrate that treatment of the As-T cells with TG upregulated the expression of PPARγ and FOXA1 whereas treatment with PD decreased the expression of some of the basal keratins. However, a combined treatment of TG and PD resulted in a consistent decrease of several proteins associated with the basal subtype of bladder cancers (KRT1, KRT14, KRT16, P63, and TFAP2A). Our data suggests that activation of PPARγ while inhibiting cell proliferation facilitates the regulation of genes involved in maintaining the luminal subtype of UC. In vivo animal studies are needed to address the efficacy of using PPARγ agonists and/or proliferation inhibitors to reduce tumor grade/stage of MIUC.

Topics: Animals; Arsenites; Biomarkers, Tumor; Cell Line, Tumor; Cell Proliferation; Down-Regulation; ErbB Receptors; Hepatocyte Nuclear Factor 3-alpha; Humans; Keratins; Mice; Mice, Nude; PPAR gamma; Quinazolines; Transcription Factors; Transcriptome; Transplantation, Heterologous; Troglitazone; Up-Regulation; Urinary Bladder Neoplasms

Genetic profiling studies on muscle-invasive bladder cancers (MIBCs) have discovered molecular subtypes with different biological characteristics. Immunohistochemical (IHC) markers such as GATA3, cytokeratin (CK) 20, CK5/6, and p53 are associated with these subtypes. In this study, we investigated the biological and prognostic significance of these IHC markers in MIBCs from 91 patients who underwent radical cystectomy. High Ki-67 indices were associated with negative CK20 (p = 0.002) and diffuse CK5/6 (p = 0.001) staining. By contrast, tumors with diffuse GATA3 expression had low Ki-67 index (p = 0.006). Regarding p53, three staining patterns were associated with a high Ki-67 index: (1) complete absence, (2) diffusely strong nuclear reactivity, and (3) diffusely strong cytoplasmic staining (p < 0.001 compared with other patterns). CK5/6 and CK20 expression was typically present in a reciprocal fashion; however, diffuse GATA3 and CK5/6 coexpression was observed in 13 (14.29%) cases. Among 78 chemotherapy-naïve patients, low GATA3 staining was associated with worse recurrence-free survival in both univariate (p = 0.008) and multivariate analyses (p = 0.002). CK20, CK5/6, or p53 expression was not associated with clinical outcome. Based on our results, IHC staining for GATA3 may help risk stratification in patients with MIBC receiving radical cystectomy. In addition, the differences in Ki-67 indices suggested that aberrant p53 expression was better defined by the three aforementioned patterns, rather than percentage of nuclear staining alone.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Female; GATA3 Transcription Factor; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Proportional Hazards Models; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

We evaluated the clinical efficacy and prognosis of muscle-invasive bladder cancer according to the basal/squamous-like (BASQ) classification system based on immunohistochemical staining [CK5/6(+), CK14(+), GATA3(-), and FOXA1(-)].. One hundred patients diagnosed with muscle-invasive bladder cancer (cT2-4 N0-3 M0) were included in the study. All patients underwent radical cystectomy after transurethral removal of bladder tumor. Immunostaining was performed for CK5/6, CK14, FOXA1, and GATA3 antibodies on tissue microarray slides, and expression patterns were quantitatively analyzed using a scanning program.. The median follow-up time was 77.4 (interquartile range: 39-120.9) months. The mean age of the patients was 65.1 ± 11.2 years. FOXA1 or CK14 expression greater than 1% was respectively positively and negatively correlated with overall survival (OS; p = 0.011 and p = 0.042, respectively), cancer-specific survival (CSS; p = 0.050 for both), and recurrence-free survival (RFS; p = 0.018 and p = 0.040, respectively). For CK5/6+ and GATA3- or FOXA1- expression, 10% CK5/6+ cells were negatively correlated with OS (p = 0.032 and p = 0.039, respectively) and with RFS in combination with FOXA1- only (p = 0.050).. In this study, CK14 expression was associated with a poor prognosis. The new classification system of bladder cancer based on molecular characteristics is expected to helpful tool for the establishment of personalized treatment strategies and associated prediction of therapeutic responses.

Topics: Aged; Biomarkers, Tumor; Cystectomy; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Keratin-14; Keratins; Male; Middle Aged; Muscle Neoplasms; Neoplasms, Squamous Cell; Prognosis; Treatment Outcome; Urinary Bladder Neoplasms

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

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

Keratinising squamous cell metaplasia (KSCM) is an uncommon diagnosis in the West. Patients typically present with lower urinary tract symptoms: haematuria (visible and non-visible), dysuria, urgency and frequency. Investigation is rigid cystoscopy. Abnormal bladder wall tissue should be resected and biopsies sent for histopathology to confirm KSCM. This is a preneoplastic condition with strong associations with squamous cell carcinoma. Due to a significant lag time, annual cystoscopy with multiple biopsies is recommended.

Topics: Aged, 80 and over; Biopsy; Carcinoma, Squamous Cell; Cystoscopy; Early Detection of Cancer; Humans; Keratins; Male; Metaplasia; Time Factors; Urinary Bladder; Urinary Bladder Neoplasms

A case of combined micropapillary and plasmacytoid urothelial carcinoma (UC) of the urinary bladder is presented for a 74-year-old male who was admitted to the hospital with gross hematuria and multifocal papillary bladder tumors. Abdominal computed tomography showed a large enhancing mass on the left lateral and anterior wall of the urinary bladder, which was highly suspicious for extravesicular extension and focal extension of the anterior lesion to the pubic bone. In voided urine, cancer cells were scattered as micropapillae or nests as well as single cells on the low power view. On a higher power view, micropapillae or nests were composed of pleomorphic, high grade tumor cells with an inverted nuclear arrangement and with acinar structures occasionally identified. Single cells were discohesive and large with a thick cytoplasm and eccentrically located nuclei. Histologically, the tumor from the resected bladder showed diffusely infiltrating micropapillae or nests with a surrounding halo and dense singly-scattered plasmacytoid cells. Immunohistochemically, the cancer cells were positive for cytokeratin-7 and cytokeratin-20 but negative for S-100, leukocyte common antigen, and vimentin. At the time of radical cystectomy, severe adhesions and peritoneal metastases were found and the surgery was discontinued. The patient received systemic chemotherapy, but died of bladder cancer 14 months after surgery.

Topics: Acinar Cells; Aged; Biomarkers, Tumor; Carcinoma; Cell Nucleus; Cytoplasm; Humans; Keratins; Male; S100 Proteins; Urinary Bladder Neoplasms

-Sarcomatoid urothelial carcinoma (UCa) is a rare but aggressive variant of bladder cancer that can show diagnostic challenges even using ancillary techniques.. -To examine immunohistochemical markers in the context of sarcomatoid UCa, including those associated with epithelial-to-mesenchymal transition.. -Twenty-eight cases of sarcomatoid UCa were rereviewed. Clinical outcomes were obtained through database search. Immunohistochemistry for clinical and epithelial-to-mesenchymal transition markers was performed.. -All patients had biopsy-proven invasive UCa; 61% (17 of 28) had sarcomatoid UCa at initial diagnosis. A recognizable epithelial component(s) was present in 17 lesions. The sarcomatoid component accounted for 65% of the lesion (average), with heterologous elements present in 3 of 28 cases (11%). The morphologic spectrum of the sarcomatoid element included spindled not otherwise specified, myxoid, pseudoangiosarcomatous, and malignant fibrous histiocytoma-like undifferentiated features. The sarcomatoid component was immunoreactive for pancytokeratin (22 of 26; 85%), p63 (20 of 26; 77%), cytokeratin 903 (17 of 26; 65%), cytokeratin 7 (16 of 26; 62%), GATA3 (16 of 26; 62%), and cytokeratin 5/6 (16 of 26; 62%). STAT-6, CD31, CD34, and HMB45 were all nonreactive, whereas smooth muscle actin often showed at least focal immunoreactivity (22 of 26; 85%). Epithelial-to-mesenchymal transition markers were frequently expressed, including vimentin (26 of 26; 100%), FoxC2 (26 of 26; 100%), SNAIL (23 of 26; 88.5%), and ZEB1 (18 of 26; 69.2%). Follow-up was available for 24 patients (median, 7 months). Sixteen of 28 patients (57%) died of disease (overall mean survival, 9.1 months). The presence of myxoid or chordoid features was associated with reduced survival (P < .05).. -Sarcomatoid UCa is an aggressive form of UCa that frequently expresses epithelial-to-mesenchymal transition markers, suggesting a possible mechanism associated with aggressive tumor behavior.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Epithelial-Mesenchymal Transition; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Urinary Bladder; Urinary Bladder Neoplasms; Urothelium

We discuss the histological and immunohistochemical features of 6 cases of urothelial carcinomas of lipid cell variant and 4 cases with shadow cell differentiation, one of which showed additionally sebaceous differentiation, one of which shows additional sebaceous differentiation, from our archive cases from the last 15 years. Conventional urothelial carcinoma (UC) was seen in all lipid cell variant cases, and micropapillary carcinoma was seen in 3. The ratio of the lipid cell component was between 10% and 40% in these 6 cases. Typical histologic features of the lipid cell variant include lipoblast-like cells with a notched nuclear appearance, abundant vacuoles, an eccentric nucleus, and pagetoid spread in some areas. GATA3 and pancytokeratin AE1/AE3 immunohistochemical staining were positive in all cases. Adipophilin was positive in various degrees in 5 of the 6 lipid cell variant cases but was also positive in the case with sebaceous differentiation. α-methylacyl-CoA racemase was positive in the lipid cell areas and negative or focal weakly positive in the conventional UC areas in 4 of the 6 cases. Vimentin, S-100 protein, and PAX8 were negative in the lipid cell component. Follow-up information was available for all cases with follow-up ranging from 6 to 84 months (mean, 34 months). Four patients died of the disease. One pT4 patient who had been followed up for 6 months lives with the disease, whereas another is disease free. In conclusion, the lipid cell variant is a rare UC variant that usually presents at an advanced stage, and tumor cells are histologically similar to lipoblasts, resemble sebaceous differentiation, and show positive immunohistochemical staining with adipophilin.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Transitional Cell; Cell Differentiation; Female; GATA3 Transcription Factor; Humans; Immunohistochemistry; Keratins; Lipid Metabolism; Male; Membrane Proteins; Middle Aged; Neoplasms, Glandular and Epithelial; Perilipin-2; Racemases and Epimerases; S100 Proteins; Sebaceous Glands; Urinary Bladder Neoplasms

The aim of study was to examine the localization and distribution of cytokeratin (CK) and vimentin protein and their association with clinical outcome of the TCC patients. Expression pattern of cytokeratin and vimentin was evaluated by immunohistochemistry in TCC cases and inflammatory lesions. Cytoplasmic expression of CK was noticed in 52.17% of TCC cases and its expression was not observed in inflammatory lesions of bladder specimens. Vimentin showed expression in 69.00% cases of TCC. Significant differences were noticed in expression pattern of CK and vimentin in inflammatory lesion and Transitional Cell Carcinoma cases. Vimentin expression increased with the grade of TCC and this difference was statistically significant whereas expression of CK decreased according to the grade of TCC. Furthermore, it was also observed that expression pattern of vimentin was high in ≥55 years as compared to <55 age group patients and these differences were significant in men as compared to women. Expression pattern of CK did not show any significant relation with age and gender. Therefore, it can be concluded that cytokeratin and vimentin will be helpful markers in the early diagnosis of Transitional Cell Carcinoma/bladder carcinoma.

Topics: Adult; Biomarkers, Tumor; Carcinoma, Transitional Cell; Female; Humans; Keratins; Male; Middle Aged; Urinary Bladder Neoplasms; Vimentin

To report on the clinicopathological features of 20 cases of microcystic urothelial bladder carcinoma.. The extent of microcystic component varied from 50-100% of the specimens. The cysts were round-oval and of varying sizes; the periphery of large cysts was frequently punctuated by many smaller cysts. The cysts were lined by urothelial, low columnar cells or by a single layer of flattened epithelium of low-intermediate nuclear grade. Focal high-grade conventional urothelial carcinoma was present in eight cases. Immunohistochemistry demonstrated variable positivity for cytokeratins 7 and 20, MUC1, MUC5AC, p63 and GATA3. Extent of expression of Ki67, p53 and p27(kip1) ranged from 20-60%, 10-40% and 10-30% of cells, respectively. On follow-up, 11 patients died of disease at 11-56 months and three patients were alive with disease at 26-37 months. Univariate survival analysis showed no differences for microcystic carcinoma versus conventional urothelial carcinoma (P = 0.548).. Microcystic urothelial carcinoma may pose diagnostic difficulties, especially in limited biopsy samples, where it may be mistaken for cystitis glandularis or adenocarcinoma of the bladder. Histological features, clinical history and appropriate immunohistochemical studies should help to distinguish it from its mimics. Aggressiveness seems to be related to higher stage at diagnosis.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Transitional Cell; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Mucins; Urinary Bladder Neoplasms; Urothelium

We present the clinicopathological features of 56 cases of the nested variant of urothelial bladder carcinoma. This is an uncommon variant of bladder cancer, recognized by the current WHO classification of urologic tumors. The nested component represented 100 % of the tumor in 24 cases. The architectural pattern of the tumor varied from solid expansile to infiltrative nests characterized by deceptively bland histologic features resembling von Brunn nests. Typical features of high-grade conventional urothelial carcinoma were present in 32 cases. Most neoplastic cells had nuclei of low to intermediate nuclear grade with occasional nuclear enlargement, most frequently seen in deep areas of tumor. The nested component expressed cytokeratins 7, 20, CAM5.2, and high molecular weight (34ßE12), p63, Ki67, p53, p27, and GATA3. Tumor extension was T1 (n = 9), minimally T2 (n = 10), T2a (n = 1), T2b (n = 4), T3a (n = 8), T3b (n = 13), and T4a (n = 11). On follow-up, 36 of patients died of or were alive with disease from 2 to 80 months (mean 21 months). Four patients died of other causes. Eleven other patients remained disease free. Univariate survival analysis showed no differences for nested carcinoma compared with conventional urothelial carcinoma. As in conventional urothelial carcinoma, in nested carcinoma of the bladder pT category defined different survival groups. In summary, nested variant of urothelial bladder carcinoma is typically associated with advanced stage. In samples of limited volume, it may be misdiagnosed as proliferation of von Brunn nests or other nested-like bladder lesions, delaying definitive therapy.

Topics: Aged; Aged, 80 and over; Carcinoma, Transitional Cell; Cell Proliferation; Female; Follow-Up Studies; GATA3 Transcription Factor; Humans; Kaplan-Meier Estimate; Keratins; Male; Membrane Proteins; Middle Aged; Neoplasm Staging; Survival Rate; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

Muscle-invasive bladder carcinoma (MIBC) constitutes a heterogeneous group of tumors with a poor outcome. Molecular stratification of MIBC may identify clinically relevant tumor subgroups and help to provide effective targeted therapies. From seven series of large-scale transcriptomic data (383 tumors), we identified an MIBC subgroup accounting for 23.5% of MIBC, associated with shorter survival and displaying a basal-like phenotype, as shown by the expression of epithelial basal cell markers. Basal-like tumors presented an activation of the epidermal growth factor receptor (EGFR) pathway linked to frequent EGFR gains and activation of an EGFR autocrine loop. We used a 40-gene expression classifier derived from human tumors to identify human bladder cancer cell lines and a chemically induced mouse model of bladder cancer corresponding to human basal-like bladder cancer. We showed, in both models, that tumor cells were sensitive to anti-EGFR therapy. Our findings provide preclinical proof of concept that anti-EGFR therapy can be used to target a subset of particularly aggressive MIBC tumors expressing basal cell markers and provide diagnostic tools for identifying these tumors.

Topics: Adult; Aged; Aged, 80 and over; Animals; Autocrine Communication; Butylhydroxybutylnitrosamine; Cell Line, Tumor; Cell Proliferation; ErbB Receptors; Female; Gene Expression Regulation, Neoplastic; Hepatocyte Nuclear Factor 3-alpha; Humans; Keratins; Male; Mice; Middle Aged; Molecular Targeted Therapy; Muscles; Neoplasm Invasiveness; Phenotype; Protein Kinase Inhibitors; Signal Transduction; Survival Analysis; Transcriptome; Treatment Outcome; Urinary Bladder Neoplasms

The lipid cell variant of urothelial carcinoma is a rare variant of urinary bladder cancer, comprised of lipoblast-like cells. In this report, we describe a case of the lipid cell variant of aggressive urothelial carcinoma. A 78-year-old man was admitted to the hospital because of gross hematuria. On cystoscopy, an ulcerative lesion, non-papillary architecture, was observed in the lateral wall of the bladder. Transurethral resection was performed. Histopathological findings of the bladder tumor indicated neoplastic cells forming irregular solid nests and sheets. Lipoblast-like neoplastic cells that had eccentric nuclei and cytoplasmic vacuoles were observed, not only in the resected specimen, but also in urine samples. On mucin histochemistry, the tumor cell cytoplasm contained no neutral or acidic mucus. The lipoblast-like cells were positive for cytokeratins (AE1/AE3, CK7) and adipophilin, known as a protein associated with neutral lipid synthesis. In general, it is difficult to prove the existence of intracytoplasmic lipid in formalin-fixed paraffin-embedded materials. This is the first report in which the presence of lipid in vacuoles of the lipid cell variant has been verified by immunohistochemistry.

Topics: Adipocytes; Aged; Carcinoma, Transitional Cell; Cell Nucleus; Humans; Keratins; Lipids; Male; Membrane Proteins; Mucins; Perilipin-2; Urinary Bladder Neoplasms; Vacuoles

Sarcomatoid carcinoma of the urinary bladder is a rare entity, whose histogenesis and biological behavior remain controversial. The cytological literature on sarcomatoid carcinoma in voided urine is very scarce. Clinically, the diagnosis of this tumor can be made by computed tomography (CT), magnetic resonance imaging (MRI), cytology, and biopsy material. In this study, cytology, histopathology, and radiological imaging were employed in order to reach a diagnosis of sarcomatoid carcinoma. CT imaging showed increased thickness of the bladder wall associated to a polypoid mass. MRI showed a 4-cm sized, broadly necked polypoid mass with calcification and ulceration at the right side of the bladder wall. T2W1 imaging showed low signal. Voided urinary cytology showed a scattered cellular presentation. The tumor cells had a high nucleo- cytoplasmic ratio, with elongated cytoplasm with faint with indistinct cytoplasm border. The nucleus was oval to round, with large and irregular nucleoli and irregular nuclear membrane. These tumor cells were positive for cytokeratin (CKAE1/AE3), vimentin, p53, carcinoembryonic antigen (CEA), α1-smooth muscle actin (SMA) by the immunoperoxidase staining. Histopathology showed spindle-shaped and clumped large tumor cells with abundant cytoplasm. Mitotic figures were frequently seen and varied from area to area (50% of the tumor cells were positive for MIB1).

Topics: Actins; Aged; Carcinoembryonic Antigen; Carcinoma; Cytodiagnosis; Epithelial Cells; Female; Humans; Keratins; Magnetic Resonance Imaging; Tomography, X-Ray Computed; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Vimentin

Current clinical judgment in bladder cancer (BC) relies primarily on pathological stage and grade. We investigated whether a molecular classification of tumor cell differentiation, based on a developmental biology approach, can provide additional prognostic information. Exploiting large preexisting gene-expression databases, we developed a biologically supervised computational model to predict markers that correspond with BC differentiation. To provide mechanistic insight, we assessed relative tumorigenicity and differentiation potential via xenotransplantation. We then correlated the prognostic utility of the identified markers to outcomes within gene expression and formalin-fixed paraffin-embedded (FFPE) tissue datasets. Our data indicate that BC can be subclassified into three subtypes, on the basis of their differentiation states: basal, intermediate, and differentiated, where only the most primitive tumor cell subpopulation within each subtype is capable of generating xenograft tumors and recapitulating downstream populations. We found that keratin 14 (KRT14) marks the most primitive differentiation state that precedes KRT5 and KRT20 expression. Furthermore, KRT14 expression is consistently associated with worse prognosis in both univariate and multivariate analyses. We identify here three distinct BC subtypes on the basis of their differentiation states, each harboring a unique tumor-initiating population.

Topics: Biomarkers, Tumor; Cell Differentiation; Cell Membrane; Gene Expression Regulation, Neoplastic; Humans; Keratins; Reproducibility of Results; Risk Factors; Survival Analysis; Urinary Bladder Neoplasms; Urothelium

To evaluate the utility of p63 and high molecular weight cytokeratin in the distinction between urothelial carcinoma with prostatic stromal invasion and urothelial carcinoma with colonisation of prostatic ducts and acini which may be challenging on H&E, especially for general pathologists who may occasionally encounter these cases.. A search of surgical pathology and consultation files was made for cystoprostatectomy specimens with confirmed urothelial carcinoma with prostatic stromal invasion. Intensity for both p63 and high molecular weight cytokeratin within the tumour cells were scored as negative/weak or strong.. A total of 34 cases were identified, 23 (68%) of which had associated foci of urothelial carcinoma with colonisation of prostatic ducts and acini. Mean patient age was 68.5 years (range 44-88 years). In all cases, basal cells of benign prostatic glands showed strong staining for both p63 and high molecular weight cytokeratin. Seventeen of 34 cases (50%) of urothelial carcinoma showed no or weak expression of high molecular weight cytokeratin in the tumour cells. The other 17 cases (50%) of urothelial carcinoma showed strong expression of high molecular weight cytokeratin in the tumour cells. Fourteen of 34 cases (41%) showed negative or weak expression of p63 in tumour cells. Twenty of 34 cases (59%) showed strong expression of p63 in tumour cells. In the 14 of 34 cases (41%) and 17 of 34 cases (50%) which showed negative/weak expression of p63 and high molecular weight cytokeratin, respectively, in the tumour cells, the positive staining of the basal cells by p63 and high molecular weight cytokeratin in the benign prostatic glands and acini or those colonised by urothelial carcinoma, aided in the distinction from urothelial carcinoma with prostatic stromal invasion. In the remaining 20 of 34 cases (59%) and 17 of 34 cases (50%) in which the tumour cells showed strong expression of p63 and high molecular weight cytokeratin, respectively, larger malignant tumour cells and smaller benign basal cells of the prostatic glands and acini were highlighted with these markers, and were easily distinguishable.. Our study suggests that p63 and high molecular weight cytokeratin may be utilised in the distinction between urothelial carcinoma with prostatic stromal invasion and urothelial carcinoma with colonisation of prostatic ducts and acini.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Diagnosis, Differential; Humans; Keratins; Male; Membrane Proteins; Middle Aged; Neoplasm Invasiveness; Prostate; Stromal Cells; Urinary Bladder Neoplasms

Even though urothelial cancer is the fourth most common tumor type among males, progress in treatment has been scarce. A problem in day-to-day clinical practice is that precise assessment of individual tumors is still fairly uncertain; consequently efforts have been undertaken to complement tumor evaluation with molecular biomarkers. An extension of this approach would be to base tumor classification primarily on molecular features. Here, we present a molecular taxonomy for urothelial carcinoma based on integrated genomics.. We use gene expression profiles from 308 tumor cases to define five major urothelial carcinoma subtypes: urobasal A, genomically unstable, urobasal B, squamous cell carcinoma like, and an infiltrated class of tumors. Tumor subtypes were validated in three independent publically available data sets. The expression of 11 key genes was validated at the protein level by immunohistochemistry.. The subtypes show distinct clinical outcomes and differ with respect to expression of cell-cycle genes, receptor tyrosine kinases particularly FGFR3, ERBB2, and EGFR, cytokeratins, and cell adhesion genes, as well as with respect to FGFR3, PIK3CA, and TP53 mutation frequency. The molecular subtypes cut across pathologic classification, and class-defining gene signatures show coordinated expression irrespective of pathologic stage and grade, suggesting the molecular phenotypes as intrinsic properties of the tumors. Available data indicate that susceptibility to specific drugs is more likely to be associated with the molecular stratification than with pathologic classification.. We anticipate that the molecular taxonomy will be useful in future clinical investigations.

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Cell Adhesion; Cell Cycle; Female; Gene Expression Profiling; Genetic Markers; Humans; Keratins; Male; Mutation Rate; Receptor Protein-Tyrosine Kinases; Urinary Bladder Neoplasms

Muscle-invasive bladder cancer is associated with a high frequency of metastasis, and bone is the most common metastatic site outside the pelvis. To clarify its organ-specific characteristics, we generated a successive bone metastatic T24-B bladder cancer subline following tail vein injection of metastatic T24-L cells. Compared with parental T24-L cells, epithelial-like T24-B cells displayed increased adhesion but decreased migration or invasion abilities as well as up-regulation of cytokeratins and down-regulation of vimentin, N-cadherin and MMP2. Mechanically, phosphatidylinositol 3-kinase (PI3K)/Akt targets glycogen synthase kinase-3β (GSK3β)/β-catenin to control ZEB1 gene transcription, and then subsequently regulates the expression of cytokeratins, vimentin and MMP2. Importantly, ZEB1 is essential for bladder cancer invasion in vitro and distant metastasis in vivo, and ZEB1 overexpression was highly correlated with the expression of those downstream markers in clinical tumor samples. Overall, this study reveals a novel mechanism facilitating metastatic bladder cancer cell re-colonization into bone, and confirms the significance of mesenchymal-to-epithelial transition (MET) in formation of bone metastasis.

Topics: Animals; beta Catenin; Bone Neoplasms; Cadherins; Cell Line, Tumor; Down-Regulation; Epithelial-Mesenchymal Transition; Female; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; Homeodomain Proteins; Humans; Keratins; Matrix Metalloproteinase 2; Mice; Mice, Nude; Phosphatidylinositol 3-Kinase; Proto-Oncogene Proteins c-akt; Signal Transduction; Transcription Factors; Transcription, Genetic; Transplantation, Heterologous; Up-Regulation; Urinary Bladder Neoplasms; Vimentin; Wnt Proteins; Zinc Finger E-box-Binding Homeobox 1

To study the clinical and pathologic characteristics of small cell neuroendocrine carcinoma of urinary tract.. All cases of urinary tract carcinoma encountered in the General Hospital of People Liberation Army during the period from 1999 to 2010 were retrospectively reviewed. The clinicopathologic data of small cell neuroendocrine carcinomas were further analyzed, with literature review.. A total of 16 cases of small cell neuroendocrine carcinoma were identified, including 10 from urinary bladder, 2 from ureter, 3 from renal pelvis, and 1 multifocal tumor involving renal pelvis and ureter. There were altogether 8 males and 8 females. The median age of the patients was 63 years (range = 24 to 79 years). Gross hematuria (11 cases) represented the main presenting symptom. Four patients had flank pain and 4 had urinary irritation symptoms. Seven patients underwent radical cystectomy. Six other patients underwent radical nephroureterectomy, 1 partial cystectomy, 1 TURBT and the remaining case biopsy only. The size of the tumor ranged from 0.8 to 8.0 cm (median = 4.5 cm). Histologically, 15 cases represented mixed small cell neuroendocrine carcinoma (with 13 mixed with transitional cell carcinoma and 2 with adenocarcinoma). Immunohistochemical study showed positive staining for neuroendocrine markers. On presentation, 1 patient was in stage pT1, 7 in stage pT2, 6 in stage pT3, 2 in stage pT4. Six patients died of the disease after operation. The overall survival was 25 months and the 5-year survival rate was 32.4%.. Small cell neuroendocrine carcinoma of urinary bladder is a highly malignant disease and associated with poor prognosis. The diagnosis relies on detailed histologic examination. Early diagnosis, when coupled with cystectomy or nephroureterectomy and adjuvant chemotherapy, represents the mainstay of management.

Topics: Adult; Aged; Carcinoma, Neuroendocrine; Carcinoma, Small Cell; CD56 Antigen; Chemotherapy, Adjuvant; Cystectomy; Female; Follow-Up Studies; Humans; Keratins; Kidney Neoplasms; Male; Middle Aged; Neoplasm Staging; Nephrectomy; Phosphopyruvate Hydratase; Retrospective Studies; Survival Rate; Synaptophysin; Ureteral Neoplasms; Urinary Bladder Neoplasms; Urologic Neoplasms; Young Adult

Reactive renal tubular cells show features of an atypical repair reaction. Differentiation between reactive renal tubular cells and low-grade urothelial carcinoma (LG-UC) cells can therefore be a diagnostic challenge based on morphology alone. In this study, we evaluated the diagnostic utility of vimentin and a high-molecular-weight cytokeratin antibody (clone 34ßE12) in differentiating reactive renal tubular cells from LG-UC.. We evaluated voided urine cytology and surgical specimens from 40 patients with renal disease, and 17 patients with LG-UC. All slides were stained with vimentin and 34ßE12.. In the reactive renal tubular cells in voided urine cytology, vimentin showed strong cytoplasmic staining in 39/40 (97.5%) cases, but all were negative for 34ßE12. LG-UC cells showed positive staining for 34ßE12 in 3/17 (17.6%) cases, whereas none were positivity for vimentin. The reactive renal tubular cells of histological specimens in the renal disease group demonstrated positive for vimentin in all 40 cases and all were negative for 34ßE12. The LG-UC group showed abnormal staining for 34ßE12 in 4/17 (23.5%) cases, whereas none were positive for vimentin.. Vimentin expression in urine cytology can help to distinguish reactive renal tubular cells from LG-UC. However, 34ßE12 does not appear to be a useful adjunct to distinguish these two groups in voided urine cytology.

Topics: Biomarkers, Tumor; Carcinoma; Cytodiagnosis; Diagnosis, Differential; Humans; Keratins; Kidney Neoplasms; Kidney Tubules; Neoplasm Staging; Urinary Bladder Neoplasms; Urothelium; Vimentin

Besides worse prognosis of bladder cancer with squamous differentiation (pure squamous cell carcinoma (SCC) or mixed urothelial carcinoma (UC/SCC)), high-grade non-keratinising squamous differentiation is difficult to identify in haematoxylin-eosin stainings. This study aims to validate routine immunohistochemical markers for squamous differentiation in a larger cohort of patients. Tissue microarrays of 89 pure SCCs and mixed UC/SCCs, 66 urothelial carcinomas (UC), precursor lesions and normal urothelium were stained for cytokeratin (CK) 5/6, CK 5/14, CK 7, CK 20 and uroplakin III. Electron microscopy was performed to confirm the differentiation. Pure SCCs displayed staining throughout the epithelium for CK 5/6 (76.6% (36/47)) and CK 5/14 (95.8% (46/48)), focal staining for CK 7 (28.9% (13/45)) and no staining for CK 20 and uroplakin III (both 0% (0/48)). UCs exhibited a basal or diffuse staining for CK 5/6 (30.2% (16/53)) and CK 5/14 (57.1% (32/56)), focal positivity for CK 7 (83.6% (46/55)), CK 20 (50.9% (29/57)) and uroplakin III (21.8% (12/55)). Each marker discriminated SCC and UC significantly (p < 0.01). A third subgroup rarely showed full epithelial staining for CK 5/6 (14.3% (1/7)) and CK 5/14 (28.6% (2/7)), focal staining for CK 7 (85.7% (6/7)) and no staining for CK 20 and uroplakin III (both 0% (0/7)). Electron microscopy could prove both, SCC and UC characteristics, revealing a transient type. A staining pattern with CK 5/6- and CK 5/14-positivity plus CK 20- and uroplakin III-negativity identified squamous differentiation in bladder tumours and revealed a third type of squamous transdifferentiation.

Topics: Biomarkers, Tumor; Carcinoma in Situ; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cell Count; Cell Differentiation; Cystectomy; Humans; Keratins; Phenotype; Prognosis; Tissue Array Analysis; Urinary Bladder Neoplasms; Urothelium

A 66-year-old woman visited our hospital complaining of painful, irritative urinary symptoms and macroscopic hematuria. Cystoscopy revealed a non-papillary tumor covered with necrotic tissue on the right side of the posterior wall of the bladder. Transurethral resection was performed ; histologically, the tumor was found to be composed of carcinomatous and sarcomatous elements. The carcinomatous element consisted of urothelial and squamous cell carcinomas. The sarcomatous element was composed of osteosarcoma, chondrosarcoma and spindle cell sarcoma. Immunohistochemical examination showed that the carcinomatous component was positive for cytokeratin and the sarcomatous component was positive for S-100 protein. The patient underwent total cystectomy with ileal conduit under the diagnosis of carcinosarcoma. Pathological examination showed no residual tumor. She was followed up with no signs of recurrence or metastasis. Computed tomography (CT) at nine months following surgery showed no evidence of recurrence. However, thirteen months after the operation, she complained of lower abdominal pain, and CT demonstrated a bulky intrapelvic tumor and right hydronephrosis. Her condition worsened rapidly and she died one month later.

Topics: Aged; Carcinosarcoma; Cystectomy; Female; Histocytochemistry; Humans; Keratins; Neoplasm Recurrence, Local; S100 Proteins; Urinary Bladder Neoplasms; Urinary Diversion

Adult rhabdomyosarcoma (RMS) in the urinary bladder is rare, and is the subject of case reports and small series. It consists of sheets of small round blue cells with high nuclear cytoplasmic ratio, brisk mitosis and apoptosis. In this study, we reported one case of pure rhabdomyosarcoma and two cases of urothelial carcinomas with extensive rhabdomyosarcomatous differentiation. In addition, their immunohistochemical profile was compared to that of small cell carcinoma of the bladder. Our study showed that sufficient sampling was critical for the diagnosis of urothelial carcinoma with extensive rhabdomyosarcomatous differentiation. As adult RMS in the bladder and urothelial carcinoma with rhabdomyosarcomatous differentiation shared morphological features with small cell carcinoma of the bladder, appropriate immunohistochemical stains were necessary in the differential diagnosis. We showed both rhabdomyosarcoma and rhabdomyosarcomatous areas of the urothelial carcinoma were positive for myogenin, negative for cytokeratin and chromogranin stains. In contrast, small cell carcinoma was positive for cytokeratin, and 7 out of 9 cases were also positive for chromogranin. Both rhabdomyosarcoma and small cell carcinoma could be positive for synaptophysin, a potential pitfall to avoid. In addition, all of the tumors with rhabdomyosarcomatous differentiation were negative for FKHR rearrangement.

Topics: Biomarkers, Tumor; Carcinoma, Small Cell; Cell Differentiation; Chromogranins; Diagnosis, Differential; Female; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratins; Male; Middle Aged; Myogenin; Predictive Value of Tests; Rhabdomyosarcoma; RNA-Binding Protein EWS; Synaptophysin; Translocation, Genetic; Urinary Bladder; Urinary Bladder Neoplasms; Urothelium

Epigenetic alterations are common and can now be addressed in a parallel fashion. We investigated the methylation in bladder cancer with respect to location in genome, consistency, variation in metachronous tumors, impact on transcripts, chromosomal location, and usefulness as urinary markers.. A microarray assay was utilized to analyze methylation in 56 samples. Independent validation was conducted in 63 samples by a PCR-based method and bisulfite sequencing. The methylation levels in 174 urine specimens were quantified. Transcript levels were analyzed using expression microarrays and pathways were analyzed using dedicated software.. Global methylation patterns were established within and outside CpG islands. We validated methylation of the eight tumor markers genes ZNF154 (P < 0.0001), HOXA9 (P < 0.0001), POU4F2 (P < 0.0001), EOMES (P = 0.0005), ACOT11 (P = 0.0001), PCDHGA12 (P = 0.0001), CA3 (P = 0.0002), and PTGDR (P = 0.0110), the candidate marker of disease progression TBX4 (P < 0.04), and other genes with stage-specific methylation. The methylation of metachronous tumors was stable and targeted to certain pathways. The correlation to expression was not stringent. Chromosome 21 showed most differential methylation (P < 0.0001) and specifically hypomethylation of keratins, which together with keratin-like proteins were epigenetically regulated. In DNA from voided urine, we detected differential methylation of ZNF154 (P < 0.0001), POU4F2 (P < 0.0001), HOXA9 (P < 0.0001), and EOMES (P < 0.0001), achieving 84% sensitivity and 96% specificity.. We initiated a detailed mapping of the methylome in metachronous bladder cancer. Novel genes with tumor, chromosome, as well as pathway-specific differential methylation in bladder cancer were identified. The methylated genes were promising cancer markers for early detection of bladder cancer.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Chromosomes, Human, Pair 21; CpG Islands; DNA Methylation; Female; Genes, Neoplasm; Humans; Keratins; Male; Methylation; Middle Aged; Oligonucleotide Array Sequence Analysis; Urinary Bladder Neoplasms

This laboratory has generated a series of seven cadmium (Cd(+2))- and six arsenite (As(+3))-transformed urothelial cancer cell lines by exposure of parental UROtsa cells to each agent under similar conditions of exposure. In this study, the seven Cd(+2)-transformed cell lines were characterized for the expression of keratin 6, 16, and 17 while the six As(+3) cell lines were assessed for the expression of keratin 7 and 19. The results showed that the series of Cd(+2)-transformed cell lines and their respective transplants all had expression of keratin 6, 16, and 17 mRNA and protein. The expression of keratin 6, 16, and 17 was also correlated with areas of the urothelial tumor cells that had undergone squamous differentiation. The results also showed that four of the six As(+3)-transformed cell lines had expression of keratin 7 and 19 mRNA and protein and produced subcutaneous tumors with intense focal staining for keratin 7 and 19. The other two As(+3)-transformed cell lines had very low expression of keratin 7 mRNA and protein and produced subcutaneous tumors having no immunoreactivity for keratin 7; although keratin 19 expression was still present. The peritoneal tumors produced by one of these two cell lines regained expression of keratin 7 protein. The present results, coupled with previous studies, indicate that malignant transformation of UROtsa cells by Cd(+2) or As(+3) produce similar patterns of keratin 6, 7, 16, 17, and 19 in the resulting series of cell lines and their respective tumors.

Topics: Animals; Arsenites; Biomarkers, Tumor; Blotting, Western; Cadmium; Cell Line; Cell Transformation, Neoplastic; Gene Expression Regulation, Neoplastic; Humans; Keratins; Mice; Mice, Nude; Neoplasm Transplantation; Real-Time Polymerase Chain Reaction; RNA, Messenger; Transplantation, Heterologous; Urinary Bladder Neoplasms; Urothelium

Topics: Carcinoma, Renal Cell; Carcinoma, Transitional Cell; Carcinosarcoma; Female; Follow-Up Studies; Humans; Keratins; Kidney Neoplasms; Lymphatic Metastasis; Male; Prostatic Neoplasms; Retrospective Studies; Survival Rate; Ureteral Neoplasms; Urinary Bladder Neoplasms; Urologic Neoplasms; Vimentin

Small cell neuroendocrine carcinoma (SCNEC) of the urinary bladder is a rare but aggressive neoplasm that usually exhibits neuroendocrine differentiation. Here, the authors report a case of SCNEC in an 80-year-old man. The patient had gross hematuria and nodular mass involving the wall of the urinary bladder. Total cystectomy was done. The tumor consisted of small, uniform, round, and spindled-shaped cells with chromatin dark nuclei and numerous mitotic figures. The cells were reactive for chromogranin, neuron-specific enolase (diffuse), and keratin (focal). Ultrastructural studies revealed neurosecretory granules and intermediate filaments. The diagnosis of SCNEC with focal high-grade urothelial component was established. No metastasis was found at the time of diagnosis and the patient refused further chemotherapy or radiotherapy. The histogenesis, differential diagnosis, and prognosis of SCNEC of the urinary bladder were discussed.

Topics: Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Neuroendocrine; Chromogranins; Cystectomy; Cytoplasmic Granules; Humans; Intermediate Filaments; Keratins; Male; Neurosecretory Systems; Phosphopyruvate Hydratase; Urinary Bladder Neoplasms

We present very rare variants of urothelial carcinoma featuring nested, plasmacytoid, and lipoid cell morphology in an 80-year-old female who was admitted to our hospital with mictritional pain and bilateral hydronephrosis. Abdominal computed tomography showed diffuse thickening of the urinary bladder, indicating probable invasion into the surrounding adipose tissue. Cytological tests on a urine specimen revealed medium-sized cancer cell clusters with hyperchromatic nuclei and dense cytoplasm, small clusters of dyshesive cells with eccentric or crescent nuclei, and abundant or vacuolated cytoplasm mimicking plasma cells or lipoblasts. Histopathological findings of the bladder tumor revealed that the nested urothelial carcinoma variants were mainly located in the lamina propria, whereas the plasmacytoid and lipoid-cell variants had deeply infiltrated into cells with a myxoid background. Immunohistochemically, the cancer cells were positive for cytokeratin, epithelial membrane antigen, but not for vimentin, S-100, or hematopoietic markers of plasma cells such as CD79alpha and CD38. The patient received no radiosurgical therapies because of the advanced stage of her disease and died a few months after the initial diagnosis. To our knowledge, this is the first cytological and histological examination of urothelial carcinoma consisting of nested, plasmacytoid, and lipoid-cell variants of the urinary bladder.

Topics: Aged, 80 and over; Carcinoma, Transitional Cell; Diagnosis, Differential; Fatal Outcome; Female; Humans; Keratins; Urinary Bladder Neoplasms

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

Involvement of the urinary bladder by prostatic adenocarcinoma (PCA) occasionally occurs. In this study, we analyzed urine cytological findings in patients with secondary involvement of the urinary bladder by PCA with the help of the immunocytochemistry. The cases were divided into two groups: (1) prospective study group: three cases; and (2) retrospective study group: 12 cases which were retrieved from our cytopathological files. The urine cytology specimens (cytospins) from all cases were submitted for prostatic specific antigen (PSA) immunocytochemistry. Additional immunostaining for high-molecular-weight cytokeratin (HMWCK) was performed if PSA immunoreactivity was negative. All cytospin smears showed atypical cells characterized by large, round and uniform nuclei with prominent nucleoli and dense cytoplasm. They were present as single cells or in cell groups simulating urothelial carcinoma. The diagnosis of PCA was made if the atypical cells were either immunoreactive for PSA or nonreactive for HMWCK. The urothelial cells were PSA- and HMWCK+. The immunostaining supported the PCA diagnosis in all three cases from the prospective group and two cases in the retrospective group. The remaining 10 cases in the retrospective group were diagnosed as negative: 3, atypia: 5 urothelial carcinoma: 2. The positive diagnosis for PCA was based on the PSA immunoreactivity or nonreactivity to HMWCK and the cytological atypia. In conclusions, immunostaining for PSA and HMWCK performed on cytospins of urine specimens from patients with a prior history of high-grade and/or stage of PCA is helpful to make a positive diagnosis of secondary bladder involvement from PCA.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Biopsy, Needle; Humans; Immunohistochemistry; Keratins; Male; Prospective Studies; Prostate-Specific Antigen; Prostatic Neoplasms; Retrospective Studies; Urinary Bladder Neoplasms; Urine

Topics: Biomarkers, Tumor; False Positive Reactions; Female; Humans; In Situ Hybridization, Fluorescence; Keratins; Male; Mass Screening; Nuclear Proteins; Point-of-Care Systems; Sensitivity and Specificity; Urinalysis; Urinary Bladder Neoplasms

The objective of this prospective study is to establish an appropriate cutoff value of urinary CYFRA 21.1 assay and to assess its utility combined with voided cytology and/or haemoglobin dipstick in the follow-up of patients with superficial bladder cancer.. From December 2000 to November 2003, 446 patients in follow-up for superficial bladder cancer (Ta-T1) after transurethral resection of the bladder (TURB) were included in a prospective study. Voided urine specimens were collected 7-14 d before cystoscopy and/or TURB for CYFRA 21.1 (one sample), haemoglobin dipstick (one sample), and cytology (three samples). All samples were processed for CYFRA 21.1 and haemoglobin dipstick according to manufacturer instructions. A control group (n=185) was obtained from patients in follow-up after transurethral resection of superficial disease (without recurrences within the following 6 mo). There were 125 recurrent transitional tumours detected by cystoscopy (34 TaG1; 53 TaG2/T1G1-2; 23 Ta-1G3/Tis, and 15 T2-4). Receiver operator characteristic (ROC) curves were constructed and cutoff values were chosen. Sensitivity, specificity, PPV (positive predictive value), NPV (negative predictive value), and their 95% confidence intervals were calculated.. ROC curve analysis based on the previously reported cutoff value of 4ng/ml for CYFRA 21.1 demonstrated a sensitivity and specificity of 43% and 68%, respectively. At a cutoff value of 1.5ng/ml, sensitivity was 73.8% with a low specificity (41%). Further lowering of the cutoff point below 1.5ng/ml did not demonstrate a significant increase in sensitivity. Therefore, this value was chosen as the most sensitive CYFRA 21.1 cutoff point during the rest of the study. Specificity increased when all the patients treated with pelvic radiotherapy or with UTI, urethral catheterisation, and intravesical instillations within 3 previous months were not included in our analysis. CYFRA 21.1 plus cytology and the combination of CYFRA 21.1, cytology, and haemoglobin dipstick demonstrated the highest overall sensitivities, and detected 91.3% of Ta-1G3 tumours and 93.3% of T2-4 tumours. However, there were one muscle-invasive tumour, two T1G3/Tis, three T1G2, and nine T1G1 neoplasms with negative combination of cytology and CYFRA 21.1 (1,5ng/ml). All these tumours were smaller than 2cm in size; most were single tumours. Nevertheless, there were 16 tumours larger than 0.5cm (0.5-2cm), and multiple neoplasms were endoscopically detected in 14 patients. Similar results were obtained through the combination of CYFRA 21.1 (cutoff: 1.5ng/ml), cytology, and haemoglobin dipstick.. In our experience the low sensitivity of urinary CYFRA 21.1, even using lower cutoff values and/or a combination with cytology and/or haemoglobin dipstick, makes its application not very useful as a surveillance tool for superficial bladder carcinoma.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Cystoscopy; Cytodiagnosis; Hemoglobinometry; Humans; Keratin-19; Keratins; Neoplasm Recurrence, Local; ROC Curve; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urine

Topics: Adult; Aged; Aged, 80 and over; Female; Humans; Keratins; Male; Middle Aged; Therapeutic Irrigation; Urinary Bladder; Urinary Bladder Neoplasms

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

CYFRA21-1, telomerase and vascular endothelial growth factor (VEGF) are regarded as useful tumor markers in the detection of bladder transitional cell carcinoma. However, the sensitivity of each single marker seemed to be unsatisfactory. In the current study, we assessed the sensitivity of the combined assay of these three markers in the detection of human bladder transitional cell carcinoma.. Voided urine samples of 100 patients with bladder transitional cell carcinoma were obtained and each sample was aliquoted for the various assay. Urinary CYFRA21-1 and VEGF were detected by enzyme-linked immunosorbent assay and telomerase detected by the telomeric repeat amplification protocol. The sensitivity of combined assay was compared to that of each single marker and urinary cytology, respectively.. In the 100 patients, the sensitivity was 74% for urinary CYFRA21-1, 71% for telomerase, 69% for VEGF, and 38% for cytology. CYFRA21-1, telomerase and VEGF proved significantly more sensitive than cytology, respectively (P = 0.000). The sensitivity of the combined assay in the current study was 94% and it was significantly higher than that of cytology, urinary telomerase, CYFRA21-1 or VEGF (P = 0.000). In 50 patients with hematuria but without bladder cancer, the overall specificity of the assays was 78% for CYFRA21-1, 84% for telomerase, 88% for VEGF, and 92% for cytology.. Combined assay of the markers which show different characteristics of the tumor behaviors seemed to be of interest in the detection of bladder transitional cell carcinoma.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Female; Humans; Keratin-19; Keratins; Male; Middle Aged; Sensitivity and Specificity; Telomerase; Urinary Bladder Neoplasms; Vascular Endothelial Growth Factor A

Some patients who have had prior bladder biopsies or transurethral resections undergo a repeat resection within several months for various reasons. The detection of a few residual tumor cells in bladder specimens with prior biopsy site changes can be challenging based on histology alone. Immunohistochemistry for cytokeratins may be used as an adjunct in this situation. We have noted several cases in which keratin stains were performed and positive cells were noted, raising the issue as to whether the cytokeratin positive cells were residual tumor cells or stromal cells. Immunohistochemistry for a panel of antibodies [AE1/AE3, CAM 5.2, high molecular weight cytokeratin, smooth muscle actin (SMA), desmin, and anaplastic lymphoma kinase (ALK)] was performed on 29 cases of bladder biopsies with prior biopsy site changes. Of 29 patients, 25 had a prior history of bladder tumor: 17 had invasive high-grade urothelial carcinoma (T1, 5 cases; T2, 11 cases; T3,1 case); 7 had noninvasive high-grade papillary urothelial carcinoma; 1 had noninvasive low-grade papillary urothelial carcinoma). One of the patients with noninvasive high-grade papillary urothelial carcinoma and one of the patents with invasive high-grade urothelial carcinoma had associated carcinoma in-situ. Four patients had prior benign bladder diagnoses: cystitis cystica et glandularis; polypoid cystitis; follicular cystitis; and neurogenic bladder with benign prostate hyperplasia. Of the 29 cases, 6 (21%) had cells with staining for at least 2 of the cytokeratin markers. Cytokeratin (CK) AE1/ AE3 was positive for cells in 8/29 cases (28%). In 6 of these cases, cells displayed a spindle cell and 2 cases a more epithelioid morphology. CAM 5.2 was positive in cells in 5/29 cases (17%); 3 of the cases had spindle cell and 2 cases epithelioid morphology. High molecular weight cytokeratin was expressed in cells in 2/29 cases (7%) with 1 case having spindle cell and 1 epithelioid morphology. SMA was positive in cells with a spindle cell morphology and negative in the more epitheloid cytokeratin positive cells. Desmin was positive in 3/6 keratin positive spindle cells and negative in keratin positive epithelioid cells. ALK was negative in all the cases. Three cases with spindle cell morphology and positivity for at least 1 of the keratins and SMA stains were interpreted as aberrant keratin expression in myofibroblastic cells based on the staining and the morphology of the spindle cells. Another 3 cases with co

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy; Carcinoma, Transitional Cell; Diagnostic Errors; Humans; Immunohistochemistry; Keratins; Middle Aged; Neoplasm Recurrence, Local; Neoplasm, Residual; Urinary Bladder; Urinary Bladder Neoplasms; Urothelium

This prospective study is the first immunocytochemical investigation of the frequency and prognostic value of CK+ tumour cells in the bone marrow of patients with transitional cell carcinoma (TCC).. Bone marrow aspirates from 228 TCC patients were taken preoperatively. Cytospins were made and stained by immunocytochemistry using the monoclonal antibodies CK2 and A45-B/B3. 27 patients with no evidence of any malignant disease served as control group.. CK+ tumour cells were detected in 28% (63/228) of the TCC patients. No CK+ cells (0/27) were detected in the control group. In multivariate analysis the detection of > or =3 CK+ cells in bone marrow was an independent prognostic factor (hazard ratio=2.7, p<0.05) in patients with T2-4 tumour classification.. Disseminated CK+ cells play a role in the biology of tumour spread of TCC, and their immunocytochemical detection can be useful in assessing the prognosis of TCC patients with an invasive tumour.

Topics: Adult; Aged; Aged, 80 and over; Antibodies; Bone Marrow Examination; Bone Marrow Neoplasms; Carcinoma, Transitional Cell; Case-Control Studies; Female; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Male; Middle Aged; Prognosis; Prospective Studies; Urinary Bladder Neoplasms

Topics: Biomarkers, Tumor; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Chromosome Aberrations; Diagnosis, Differential; Humans; Keratins; Lymphoma; Mucin-1; Urinary Bladder; Urinary Bladder Neoplasms

Topics: Aged; Biomarkers; Carcinoma, Transitional Cell; Female; Humans; Keratin-20; Keratin-7; Keratins; Male; Plasma Cells; Urinary Bladder Neoplasms

A clinicopathologic study of 108 cases of high-grade urothelial carcinomas of the renal pelvis is presented. Of the 108 tumors, 44 (40%) showed unusual morphologic features, including micropapillary areas (four cases), lymphoepithelioma-like carcinoma (two cases), sarcomatoid carcinoma (eight cases, including pseudoangiosarcomatous type), squamous differentiation and squamous cell carcinoma (15 cases), clear cells (two cases), glandular differentiation (two cases), rhabdoid, signet-ring or plasmacytoid cells (four cases), pseudosarcomatous stromal changes (four cases) and intratubular extension into the renal pelvis (three cases). Pathological staging was available in 62 patients; of these, 46 cases (74%) were in high stage (pT2-pT4) and 16 (26%) were in low stage (pTis, pTa, pT1). Clinical follow-up ranging from 1 to 256 months (median: 50 months) was available in 42 patients; of these, 26 (61%) died of tumor with a median survival of 31 months. The patients who did not die of their tumors showed only minimal or focal infiltration of the renal parenchyma by urothelial carcinoma, whereas those who died of their tumors showed massive infiltration of the kidney by the tumor. High-grade urothelial carcinomas of the renal pelvis can show a broad spectrum of histologic features similar to those seen in the urinary bladder. Our results support the finding that, unlike urothelial carcinomas of the bladder, the majority of primary urothelial carcinomas of the renal pelvis are of high histologic grade and present in advanced stages. Our study further highlights the fact that, in the renal pelvis, urothelial carcinomas show a tendency to frequently display unusual morphologic features and metaplastic phenomena. The importance of recognizing these morphologic variants of urothelial carcinoma in the renal pelvis is to avoid confusion with other conditions. The possibility of a high-grade urothelial carcinoma should always be considered in the evaluation of a tumor displaying unusual morphologic features in the renal pelvis, and attention to proper sampling as well as the use of immunohistochemical stains will be of importance to arrive at the correct diagnosis.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Renal Cell; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; In Situ Hybridization; Keratin-7; Keratins; Kidney Neoplasms; Kidney Pelvis; Male; Middle Aged; RNA, Viral; Urinary Bladder Neoplasms

This study addresses the diagnostic value of the Nuclear Matrix Protein 22 (NMP22) test and the Urinary Bladder Cancer (UBC II) test, in comparison to bladder wash cytology for the detection of early recurrence of bladder cancer. Patients with transitional cell carcinoma of the bladder (TCC, n = 60) and patients with benign urological diseases (n = 30) were included in this study. Voided urine samples were divided into 2 aliquots: aliquot 1 was assayed for NMP22 and aliquot 2 was tested for UBC II. Saline bladder washings were used for cytologic examination. Urine samples from TCC patients were collected before transurethral resection and on postoperative day 10. On day 10, 15 NMP22 results and 7 UBC II results exceeded the normal ranges; 4 of the cytology samples were positive for malignancy. Based on cystoscopic findings at 3 mo post-resection, 21 of the cases were classified as early recurrence; 11 of the early recurrences had been in the elevated NMP22 group, 4 in the elevated UBC II group, and 3 in the positive cytology group at 10 days post-resection. The NMP22 test gave the highest sensitivity for detecting early recurrent tumors (11 of 21, 52%). Such high sensitivity did not occur with the UBC II test (4 of 21, 19%) or cytology (3 of 21, 14%). These differences were significant (p = 0.024 and 0.009, respectively). Thus, the NMP22 test showed superiority over the other tests for detection of early recurrence of bladder cancer.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Middle Aged; Neoplasm Recurrence, Local; Nuclear Proteins; Prognosis; ROC Curve; Sensitivity and Specificity; Urinary Bladder Neoplasms

A simple and non-invasive methods for the diagnosis of transitional cell carcinoma of the bladder are needed for the prevention of invasive tumor. A proteomic technology has recently been developed to facilitate protein profiling of biological mixtures. We tried to detect the marker proteins by proteomic approach during the initiation stages on N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced bladder carcinogenesis in rats.. Ten rats in group A were given 0.05% BBN in drinking water for 12 weeks. Other 10 rats in group B with 10 rats were designated as a control group and were not given BBN. Whole urinary bladders of all rats were excised at 12 weeks from the beginning of the experiment. Conventional proteomics was performed with high resolution 2-dimension gel electrophoresis followed by computational image analysis and protein identification using mass spectrometry.. A comparison of urinary bladder hyperplasia tissue with control tissue showed that five proteins; actin gamma2 propeptide, cytokeratin-20 (CK-20), proapolipoprotein, alpha2 actin (alpha-cardiac actin) and heat shock 27 kDa protein-1 were over-expressed in hyperplastic tissues. Three proteins; transcription factor myocardin, seminal vesicle secretory protein VI (SVS-VI) precursor and hypothetical protein RMT-7 were under-expressed in hyperplastic tissues.. In our animal mode, BBN-induced urinary bladder mucosal hyperplasia resulted in an increase in the expression of five proteins and a decrease in the expression of three proteins. Of these, CK-20 and SVS-VI seem to be of particular interest. However other method such as Western blotting seems to be needed for confirmation of these proteins and more information on human bladder tissue is needed for clinical application.

Topics: Animals; Biomarkers; Butylhydroxybutylnitrosamine; Keratin-20; Keratins; Male; Precancerous Conditions; Proteomics; Rats; Rats, Sprague-Dawley; Seminal Vesicle Secretory Proteins; Urinary Bladder Neoplasms

Pseudosarcomatous myofibroblastic proliferation of the genitourinary tract is rare and may develop after trauma or spontaneously. The aim of this study was to characterize further the clinicopathological features of these lesions and to examine their relationship to inflammatory myofibroblastic tumour (IMT).. Twenty-seven cases of pseudosarcomatous myofibroblastic proliferation were analysed. There were seven males and 20 females; median age was 37 years (range 16-88). Most lesions were from the bladder (n = 21), while others were in the urethra, vulva, vagina, rectum and retrovesical space. Median tumour size was 30 mm (range 6-120 mm). Seven cases (25%) had a history of prior trauma or surgery. Three cases recurred locally but not destructively. The tumours had fasciitis-like features including bland spindle cells with evenly distributed chromatin, admixed inflammatory cells (mainly lymphocytes) and often a myxoid stroma. Immunohistochemistry showed positivity for smooth muscle actin in 14/20 cases, keratin in 8/19, desmin in 7/20 and anaplastic lymphoma kinase (ALK) in 10/21 cases. Fluorescent in situ hybridization was performed in six ALK+ cases; all were negative for ALK gene rearrangement.. Pseudosarcomatous myofibroblastic proliferations of the genitourinary tract may show ALK immunopositivity but do not show consistent ALK rearrangement. Given subtle morphological differences and more consistently benign behaviour, their relationship to inflammatory myofibroblastic tumour at other sites remains uncertain.

Topics: Actins; Adolescent; Adult; Aged; Aged, 80 and over; Anaplastic Lymphoma Kinase; Desmin; Female; Fibroblasts; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratins; Male; Middle Aged; Muscle, Smooth; Myofibroma; Protein-Tyrosine Kinases; Receptor Protein-Tyrosine Kinases; Sarcoma; Urethral Neoplasms; Urinary Bladder Neoplasms

There is evidence that carcinoma in situ (CIS) is the precursor of invasive urothelial carcinoma, a tumour characterized by frequent gene promoter methylation. The timing of altered DNA methylation is unknown in this pathway. Here we investigate gene methylation in 196 consecutive samples of normal urothelium, CIS, and tumours from 104 patients with both CIS and invasive urothelial carcinoma using quantitative methyl-sensitive polymerase chain reaction for six genes (p16, p14, E-cadherin, RARbeta2, RASSF1a, and GSTP1). Control normal urothelial samples from 15 patients with no history of urothelial carcinoma were also analysed. Immunohistochemistry established the expression of well-characterized CIS markers p53 and cytokeratin 20. Promoter methylation occurred frequently in both normal urothelium and CIS samples from patients with urothelial carcinoma, and increased with progression from normal to invasive urothelial carcinoma, at both specific loci (chi2 test: E-cadherin, p=0.0001; RASSF1a, p=0.003, RARbeta2, p=0.007, p16, p=0.024) and in general (methylation indices [t-test, p<0.0001]). Methylation was associated with cytokeratin 20 expression (t-test, p=0.004) and poor prognosis, and with increased progression to tumour death in patients whose CIS samples showed methylation, in comparison with those without methylation (log rank p<0.03). Promoter methylation occurs early in the urothelial carcinogenic pathway and appears to be a good biomarker of the invasive urothelial carcinoma phenotype.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Disease Progression; DNA Methylation; DNA, Neoplasm; Female; Humans; Keratin-20; Keratins; Male; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Polymerase Chain Reaction; Promoter Regions, Genetic; Survival Analysis; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

We have developed an office-based dot-EIA for the detection of a urinary high molecular weight cytokeratin (CK). Immunohistochemical staining and western blot based on CK1K10 monoclonal antibody were used to identify the CK. Urine of 192 patients with different types, grades, and stages of bladder tumor and 72 controls were evaluated using dot-EIA. An intense and diffuse cytoplasmic reaction was shown in bladder squamous cell carcinoma. The target epitope was identified in urine at 65, 56, and 40-kDa. The CK purified from urine showed single polypeptide at 65-kDa using SDS-PAGE and single peak at 7.4 min using capillary zone electrophoresis. The dot-EIA detected the CK with high sensitivity (97%) and specificity (94%). The CK was not detected in urine of bladder cancer patients showing response to radiotherapy. The sensitive and specific office-based detection of urinary cytokeratin would be helpful in rapid diagnosis and follow up of bladder carcinoma.

Topics: Adult; Aged; Antibodies, Monoclonal; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Molecular Weight; Neoplasm Staging; Urinary Bladder Neoplasms

An optimal immunohistochemical panel to distinguish poorly differentiated prostate (PCa) from urothelial (UCa) carcinoma was selected from a panel consisting of prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP), high-molecular-weight cytokeratin (HMWCK), clone 34betaE12, cytokeratin (CK) 7, CK20, p63, and alpha-methylacyl-coenzyme A racemase. The pilot group was composed of poorly differentiated UCa (n = 36) and PCa (n = 42). PSA and PAP stained 95% of PCa vs 0% and 11% of UCa cases, respectively. HMWCK and p63 stained 97% and 92% of UCa vs 2% and 0% of PCa cases respectively. CK7/CK20 coexpression was noted in 50% of UCa cases, whereas 86% of PCa cases were negative with both. A panel of PSA, HMWCK, and p63 was optimal for separating 95% PCa (PSA+/HMWCK and/or p63-) vs 97% UCa (PSA-/HMWCK and/or p63+). This panel was used on 26 diagnostically challenging cases and resolved 81% of cases as UCa vs PCa. The majority of PCa cases retain PSA. Negative PSA with positive HMWCK and/or p63 establishes a diagnosis of UCa.

Topics: Adenocarcinoma; Biomarkers, Tumor; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Male; Membrane Proteins; Molecular Weight; Pilot Projects; Prostate-Specific Antigen; Prostatic Neoplasms; Urinary Bladder Neoplasms; Urothelium

The Urinary Bladder Cancer (UBC) test is a marker that detects urinary fragments of cytokeratin 8 and 18. The aim of this study is to evaluate the usefulness of the pre and post operative UBC test to detect early recurrences of a bladder tumor in the first year after the transurethral resection of a bladder tumor.. A multicentric perspective study on 36 patients with superficial bladder cancer (pTa-pT1) treated with transurethral resection (TUR) was performed. Each patient underwent 4 specific urine collections: 1) preoperatively, 2) 3 days after TUR, 3) 7 days after TUR, 4) 30 days after TUR. UBC was analysed on urine with the IRMA method and the cut off value of 12 mg/L was used. Cystoscopy was performed after 3, 6, 9, and 12 months after TUR, with the aim of identifying all cancer recurrences in the first year postoperatively. Statistical analyses to identify differences between patients with or without early recurrence were performed in accordance with Fisher's exact test and Chi-square analysis.. Of the 36 patients included in the study 15 showed early recurrence and 21 were recurrence free 1 year after surgery. UBC levels measured in recurrence free patients 30 days after TUR showed normal values, values decreasing as compared with preoperative levels or both circumstances, even if a statistically significant difference was not found between the two groups.. In this study we reported an insignificant correlation between the postoperative modifications of UBC levels and the risk of tumor recurrence during the first year of follow-up. A larger study group with longer follow-ups will probably allow better evaluation of the real power of UBC tests in clinical practice.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Transitional Cell; Humans; Keratins; Neoplasm Recurrence, Local; Postoperative Care; Preoperative Care; Prospective Studies; Time Factors; Urinary Bladder Neoplasms

Topics: Adenocarcinoma, Clear Cell; Aged; Carcinoembryonic Antigen; Carcinoma, Transitional Cell; Diagnosis, Differential; Gene Expression Regulation, Neoplastic; Humans; Keratins; Male; Membrane Proteins; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

The case of an 83-year-old woman with an uncommon presentation of cutaneous metastases from muscle-invasive transitional cell carcinoma of the urinary bladder is reported. The band-like eruption of the metastatic lesion can often be misdiagnosed and treated initially as herpes zoster. A detailed immunohistochemical analysis is also described to differentiate metastatic lesions from other sources.

Topics: Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Diagnosis, Differential; Fatal Outcome; Female; Herpes Zoster; Humans; Keratins; Neoplasm Proteins; Skin Neoplasms; Urinary Bladder Neoplasms

Epithelial-to-mesenchymal transition (EMT) increases cell migration and invasion, and facilitates metastasis in multiple carcinoma types, but belies epithelial similarities between primary and secondary tumors. This study addresses the importance of mesenchymal-to-epithelial transition (MET) in the formation of clinically significant metastasis. The previously described bladder carcinoma TSU-Pr1 (T24) progression series of cell lines selected in vivo for increasing metastatic ability following systemic seeding was used in this study. It was found that the more metastatic sublines had acquired epithelial characteristics. Epithelial and mesenchymal phenotypes were confirmed in the TSU-Pr1 series by cytoskeletal and morphologic analysis, and by performance in a panel of in vitro assays. Metastatic ability was examined following inoculation at various sites. Epithelial characteristics associated with dramatically increased bone and soft tissue colonization after intracardiac or intratibial injection. In contrast, the more epithelial sublines showed decreased lung metastases following orthotopic inoculation, supporting the concept that EMT is important for the escape of tumor cells from the primary tumor. We confirmed the overexpression of the IIIc subtype of multiple fibroblast growth factor receptors (FGFR) through the TSU-Pr1 series, and targeted abrogation of FGFR2IIIc reversed the MET and associated functionality in this system and increased survival following in vivo inoculation in severe combined immunodeficient mice. This model is the first to specifically model steps of the latter part of the metastatic cascade in isogenic cell lines, and confirms the suspected role of MET in secondary tumor growth.

Topics: Animals; Blotting, Western; Carcinoma, Transitional Cell; Cell Differentiation; Cell Line, Tumor; Epithelial Cells; Gene Expression Regulation, Neoplastic; Humans; Keratins; Mesoderm; Mice; Neoplasm Metastasis; Neoplasms, Experimental; Protein Isoforms; Receptor, Fibroblast Growth Factor, Type 2; Reverse Transcriptase Polymerase Chain Reaction; Survival Analysis; Time Factors; Transplantation, Heterologous; Urinary Bladder Neoplasms; Vimentin

We present 51 cases of primary small cell carcinoma of the bladder in a clinicopathological study with emphasis on features that aid in the initial recognition and diagnosis of small cell carcinoma of the bladder.. The patients were 40 men and 11 women between the ages of 39 and 87 years (mean age 67 years). Clinical data were available in 41 cases. The most common symptomatology was haematuria in 63% of the patients while dysuria was present in 12%. Thirty-eight patients were caucasians; seven patients were Hispanics; two patients were Asian; one patient was African-American; in the three additional patients no racial information was obtained. Biopsy material was obtained in all of the patients. Cystectomy was performed in 20 patients. At diagnosis, clinical stage was as follows: stage I in two (5%), stage II in 18 (44%), stage III in 10 (24%), and stage IV in 11 (27%). Histologically, urothelial carcinoma was present in 70% of the cases, adenocarcinoma in 8%, and squamous cell carcinoma in 10% of the cases. Small cell carcinoma was the only histology present in only 12% of the cases studied. Immunohistochemical studies using chromogranin, synaptophysin and chromogranin were positive in 30-70% of the cases.. The present study highlights the unusual phenomenon of pure small cell carcinoma of the bladder and its association with other non-small cell carcinomas in that anatomical location. In addition, the study highlights the different modalities employed to treat patients in whom there is a component of small cell carcinoma of the bladder.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Chromogranins; Disease-Free Survival; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplasm Staging; Synaptophysin; Urinary Bladder Neoplasms

A 49-year-old woman underwent hysterectomy and bilateral adnexectomy after the diagnosis of a right ovarian tumor with paraaortic and pelvic lymph node metastases. The pathological diagnosis was undifferentiated carcinoma of the ovary. After the operation, a bladder tumor was discovered during the evaluation for microscopic hematuria. The bladder tumor was pathologically diagnosed as transitional cell carcinoma, pT1b, G3. Although the pathological findings of the bladder cancer and ovarian cancer were very similar, we could diagnose primary bladder cancer with ovary and lymph node metastases according to the immunohistochemical staining pattern of cytokeratins 7 and 20. Herein, the clinical usefulness of immunohistochemical staining using cytokeratins for making a differential diagnosis of the origin of a tumor in the pelvic cavity is demonstrated.

Topics: Biomarkers, Tumor; Carcinoma; Carcinoma, Transitional Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Middle Aged; Ovarian Neoplasms; Urinary Bladder Neoplasms

The inducible form of cyclooxygenase (COX), COX-2, is up-regulated in many epithelial cancers and its prostaglandin products increase proliferation, enhance angiogenesis, and inhibit apoptosis in several tissues. Pharmacologic inhibition and genetic deletion studies showed a marked reduction of tumor development in colon and skin. COX-2 has also been strongly implicated in urinary bladder cancer primarily by studies with nonselective COX- and COX-2-selective inhibitors. We now show that forced expression of COX-2, under the control of a keratin 5 promoter, is sufficient to cause transitional cell hyperplasia (TCH) in 17% and 75% of the heterozygous and homozygous transgenic lines, respectively, in an age-dependent manner. TCH was strongly associated with inflammation, primarily nodules of B lymphocytes; some T cells and macrophage infiltration were also observed. Additionally, transitional cell carcinoma was observed in approximately 10% of the K5.COX-2 transgenic mice; no TCH or transitional cell carcinoma was observed in wild-type bladders. Immunohistochemistry for vascular proliferation and vascular endothelial growth factor showed significant increases above that in wild-type urinary bladders. Our results suggest that overexpression of COX-2 is sufficient to cause hyperplasia and carcinomas in the urinary bladder. Therefore, inhibition of COX-2 should continue to be pursued as a potential chemopreventive and therapeutic strategy.

Topics: Animals; B-Lymphocytes; Carcinoma, Transitional Cell; Cell Proliferation; Cyclooxygenase 2; Gene Expression Regulation, Enzymologic; Humans; Hyperplasia; Inflammation; Keratin-15; Keratin-5; Keratins; Macrophages; Membrane Proteins; Mice; Mice, Transgenic; Neoplasm Staging; Promoter Regions, Genetic; Prostaglandin-Endoperoxide Synthases; T-Lymphocytes; Transcription, Genetic; Urinary Bladder; Urinary Bladder Neoplasms; Vascular Endothelial Growth Factor A

We report a 73-year-old male patient with leptomeningeal metastasis from urinary bladder adenocarcinoma. He was presented with prominent hyperactive delirium during the course of the disease. Meningeal carcinomatosis was detected 5 days before his death, but the primary site of the malignant tumor could not be determined. Necropsy revealed leptomeningeal infiltration of many adenocarcinoma cells that covered the cerebrum. The leptomeninges of the right middle frontal gyrus, superior temporal gyrus, precentral gyrus and inferior parietal lobe were most severely affected by tumor cell infiltration. Cerebral edema was found to extensively cover the basal part of the temporal lobe. In the cerebrum, tumor cells were clustered in the perivascular spaces and had invaded localized areas of the frontal lobe. Vascular cell adhesion molecule (VCAM)-1 expression was detected in the small vessels of the cerebral upper cortical layers and of temporal subcortical u-fibers. Numerous astrocytes positive for cytokeratin AE1/AE3 were found in the frontal and temporal lobes. Meningeal carcinomatosis from urinary bladder adenocarcinoma is extremely rare and up-regulation of the adhesion molecules in the meningeal adenocarcinoma was confirmed.

Topics: Adenocarcinoma; Aged; Astrocytes; Fatal Outcome; Humans; Immunohistochemistry; Keratins; Magnetic Resonance Imaging; Male; Meningeal Neoplasms; Urinary Bladder Neoplasms; Vascular Cell Adhesion Molecule-1

To present a pilot study to determine whether the alpha1-adrenoceptor antagonist terazosin can induce apoptosis in transitional cell carcinoma (TCC) of the bladder, similar to the effect seen with prostate cancer. The alpha1-adrenoceptor antagonist terazosin has recently been shown to induce apoptosis in prostate cancer cells both in vitro and in vivo and to reduce prostatic tissue vascularity by potentially affecting endothelial cell adhesion.. The records of 24 men who underwent radical cystectomy for TCC of the bladder at the Lexington Veterans Affairs Medical Center were reviewed. The control group consisted of 15 men who were never exposed to terazosin. The study group consisted of 9 men who were treated with terazosin before cystectomy. Sections of the bladder tumor and normal trigone were subjected to immunohistochemical analysis for microvessel density, endothelial cell CD31 expression, and apoptosis detection (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling), as well as high-molecular-weight cytokeratin staining.. A significant reduction in tissue vascularity (14.0 versus 19.2, P <0.05) and a significant increase in the apoptotic index (3.0% versus 1.7%, P <0.05) was detected in terazosin-treated bladder tumors compared with untreated bladder tumors. Most TCC specimens (80%) exhibited strong and consistently uniform immunostaining for high-molecular-weight cytokeratin staining.. These results suggest that terazosin reduces tumor vascularity and induces apoptosis in TCC of the bladder. Additional studies with more patients are necessary to reach definitive conclusions. However, considering the proven apoptotic action of terazosin in prostatic tissue, this study may have implications for the use of terazosin in the treatment of bladder TCC.

Topics: Adrenergic alpha-Antagonists; Aged; Aged, 80 and over; Antineoplastic Agents; Apoptosis; Carcinoma, Transitional Cell; Humans; In Situ Nick-End Labeling; Keratins; Male; Middle Aged; Neovascularization, Pathologic; Prazosin; Urinary Bladder Neoplasms

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

We evaluated serum levels of soluble fragments of cytokeratin 19 (CYFRA 21-1) by immunoassay (ES-700; Roche Diagnostics, Mannheim, Germany) to assess its usefulness in the diagnosis and follow-up of bladder cancer. The study included 39 patients with a diagnosis of transitional cell carcinoma (group 1) and 190 patients (group 2) with no evidence of tumor. In group 2, 180 patients had a history of bladder cancer, and 10 had benign prostatic hyperplasia. Significant differences in CYFRA 21-1 concentrations between groups 1 and 2 (P<0.01) were noted. However, CYFRA 21-1 levels did not significantly differ between the patients with benign prostatic hyperplasia and those with bladder cancer (P=0.274). CYFRA 21-1 values were higher in invasive bladder cancer compared to that detected in superficial stages (P=0.011). Setting the optimal cutoff value at 2.5 ng/mL resulted in a sensitivity of 43.6% and a specificity of 82.1%. No statistical differences were found when we compared disease-free time among the 66 patients with recurrences (30.7 months with levels <2.5 ng/mL vs. 41.2 months with levels >2.5 ng/mL; P=0.248). The risk of recurrence in patients with levels lower than 2.5 ng/mL (0.79) was no different (P=0.097) from that found in patients with higher levels (1.69). CYFRA 21-1 serum levels do not provide enough sensitivity to justify its application in routine protocols for the detection and follow-up of bladder cancer.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Female; Follow-Up Studies; Humans; Keratin-19; Keratins; Male; Middle Aged; Urinary Bladder Neoplasms

Lymphoepithelioma-like carcinoma (LELC) of the urinary bladder is very rare. It is mandatory to make differential diagnosis among lymphoma, chronic cystitis and LELC because of different therapeutic approach. A bladder tumor was found in a 90-year-old patient suffering from hematuria. After transurethral resection, undifferentiated tumor with prominent lymphoid infiltration was seen on light microscopy. Immunohistochemical examination demonstrated positive staining of tumor cells with cytokeratin (CK), epithelial membrane antigen and CK-20. We presented the case because of its rarity and related literature was reviewed.

Topics: Aged, 80 and over; Diagnosis, Differential; Humans; Immunohistochemistry; Keratin-20; Keratins; Male; Mucin-1; Urinary Bladder Neoplasms

Previous studies have shown that serum levels of the degradation products of cytokeratins could be used as surrogate markers in the diagnosis and followup of patients with solid tumors, including tumors of the bladder.. The soluble cytokeratin 19 fragment CYFRA 21-1 was measured by solid phase radioimmunoassay in the serum of 142 patients with invasive transitional cell cancer of the bladder. Of the patients 56 had clinical stage I to III locally confined disease (T1-4aN0M0) and 86 had stage IV metastatic disease with lymph node and/or distant metastases. A control group consisted of 33 healthy volunteers. In a subgroup of 49 patients with metastatic disease receiving combined platinum based chemotherapy serum CYFRA 21-1 was determined prior to the initiation of therapy and after the documentation of response.. Abnormal CYFRA 21-1 was observed in 7% of patients with locally invasive disease and in 66% of those with metastatic disease (p < 0.0001). There was no correlation of CYFRA 21-1 with tumor differentiation. Patients with abnormal CYFRA 21-1 showed statistically significant worse median overall survival. Moreover, in the subgroup of patients with metastatic disease receiving chemotherapy CYFRA 21-1 levels correlated with the response to treatment.. Patients with transitional cell cancer of the bladder with evidence of distant metastases showed a significant increase in serum CYFRA 21-1. During chemotherapy CYFRA 21-1 appears to be a potentially sensitive and useful indicator for monitoring treatment response.

Topics: Aged; Analysis of Variance; Antigens, Neoplasm; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Carcinoma, Transitional Cell; Case-Control Studies; Cohort Studies; Female; Humans; Keratin-19; Keratins; Male; Middle Aged; Monitoring, Physiologic; Neoplasm Invasiveness; Neoplasm Staging; Prognosis; Reference Values; Risk Assessment; Sensitivity and Specificity; Statistics, Nonparametric; Survival Analysis; Treatment Outcome; Urinary Bladder Neoplasms

Distinguishing between primary adenocarcinomas and secondary colonic adenocarcinomas of the urinary bladder is often difficult because they appear morphologically similar but invariably require different treatment strategies. The aim of the study was to define the utility of a limited immunohistochemical panel consisting of CDX-2, cytokeratins 7 (CK7) and 20 (CK20), and carcinoembryonic antigen (CEA) in differentiating primary from secondary bladder adenocarcinomas. Formalin-fixed, paraffin-embedded tissues from 8 primary bladder adenocarcinomas and 23 colorectal adenocarcinomas involving the bladder were included in the study. Statistical analysis was performed using the Fisher exact test. The majority (87.5%) of primary bladder adenocarcinomas were CDX-2 negative, and only one case of primary bladder adenocarcinoma was positive, while CDX-2 was strongly expressed in the nucleus of all cases of secondary (colonic) bladder tumor (P < 0.0005). Five cases (62.5%) of primary bladder adenocarcinoma and one case (4.3%) of secondary bladder tumor showed positive staining for CK7 (P = 0.002), whereas CK20 showed positive staining in five cases (62.5%) of primary bladder adenocarcinoma and in all the secondary bladder tumors (P = 0.012). All 23 secondary bladder tumors and 7 primary bladder adenocarcinomas (87.5%) expressed CEA (P = 0.25). These data demonstrate that a restricted immunohistochemical panel consisting of CDX-2, CK7, CK20, and CEA may be of use in differentiating primary bladder adenocarcinoma from secondary adenocarcinoma of colorectal origin.

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; Carcinoembryonic Antigen; CDX2 Transcription Factor; Colorectal Neoplasms; Diagnosis, Differential; Female; Homeodomain Proteins; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Trans-Activators; Urinary Bladder Neoplasms

Current methods used to determine pathological examination of the lymphatics after radical cystectomy are tedious and costly. We performed a systemic study of uroplakin II (UP II) and cytokeratin 20 (CK 20) expression in pelvic lymph nodes on multiple sides in patients with bladder cancer.. A total of 82 pelvic lymph node and 19 bladder tumor samples were obtained from 21 patients with bladder cancer by radical cystectomy with pelvic lymphadenectomy for reverse transcriptase-polymerase chain reaction assay.. Of the 19 bladder tumor tissue specimens 19 (100%) and 13 (68.4%) were positive for UP II and CK 20 mRNA expression, respectively. UP II mRNA was detected in 15 of 16 pelvic lymph node samples (93.8%) with pathologically proven metastases, whereas 9 (56.6%) were positive for CK 20 mRNA. The reverse transcriptase-polymerase chain reaction assay for UP II was statistically more sensitive than that for CK 20 in detecting not only primary tumors, but also metastatic pelvic lymph nodes (p = 0.0179 and 0.0373, respectively). Of 66 pelvic lymph node samples without metastasis UP II was detected in 6 (10%), while CK 20 was not. In addition, UP II and CK 20 mRNA could be detected in at least 50 and 500 bladder cancer HT1197 cells, respectively.. These results indicate that UP II might be a more useful marker than CK 20 for detecting micrometastases of bladder cancer in the pelvic lymph nodes, although a greater number of patients and longer followup are needed to come to a definitive conclusion.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Female; Humans; Japan; Keratin-20; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Membrane Proteins; Middle Aged; Neoplasm Staging; Predictive Value of Tests; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Urinary Bladder Neoplasms; Uroplakin II

There were investigated 80 patients with superficial urothelial carcinoma of the urinary bladder (Ta and T1). All cases had a minimum follow-up of 36 months. Patients were selected in order to form two subgroups. The first group (n = 28) had no recurrence during the follow-up, and in the second (n = 52) each patient had a minimum of two recurrences. The pathologic diagnosis was performed on archive slides stained with haematoxylin-eosin. Archive smears performed at the moment of the primary diagnosis from voided urine were discolored, and used for immunocytochemistry. It was investigated the expression of cytokeratin 20, high molecular weight cytokeratin 34betaE12, and overexpression of HER-2/neu protein. Microwave antigen retrieval was performed in all immunocytochemical procedures. The working system was LSAB2 and EnVision, and diaminobenzidine was used as chromogen. The initial conventional cytology was positive for malignant cells in all cases. Cytokeratin 20 was positive in 92.85% of cases without recurrences, and in 94.23% of cases with recurrences. High molecular weight cytokeratin was expressed in 42.85% of cases without recurrence and in 63.46% of patients with recurrences. The overexpression of HER-2/neu protein was found in 7.14% of cases of the first group, and in 84.61% of cases of the second group. Our results support the idea that HER-2/neu and high molecular weight cytokeratin may be used as markers to predict recurrence in superficial urothelial carcinoma.

Topics: Cytoplasm; Humans; Keratin-20; Keratins; Predictive Value of Tests; Receptor, ErbB-2; Recurrence; Urinary Bladder Neoplasms

Topics: Adenocarcinoma; Aged; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Transitional Cell; Diagnosis, Differential; Humans; Keratins; Male; Neoplasm Recurrence, Local; Prostatic Neoplasms; Urinary Bladder Neoplasms

We present a case of adenocarcinoma developing at the vesicocutaneous edge of a vesicostomy, 40 years after it was created, in a patient who underwent cadaveric kidney transplant. Although transitional and squamous cell carcinoma of a vesicostomy have been reported, to our knowledge, the presence of adenocarcinoma at the vesicostomy edge has not been reported previously.

Topics: Abnormalities, Multiple; Adenocarcinoma; Adult; Biomarkers, Tumor; Cell Transformation, Neoplastic; Cystostomy; Dermatologic Surgical Procedures; Female; Humans; Immunosuppression Therapy; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Kidney Failure, Chronic; Kidney Transplantation; Neoplasm Proteins; Postoperative Complications; Surgical Stomas; Time Factors; Urinary Bladder Neoplasms; Urinary Bladder, Neurogenic; Urogenital Abnormalities; Urothelium

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

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

To investigate whether prognosis in micropapillary urothelial carcinoma is related to the proportion of the micropapillary component (MPC), and to identify the immunohistochemical features of MPC.. This study presents a clinicopathological analysis of 20 patients with micropapillary urothelial carcinoma of the bladder with cystectomy specimens for evaluation. Tumours were stratified on the extent of MPC: focal, <10%; moderate, 10-50%; extensive, >50%; and this was correlated with tumour stage and prognosis. Sixteen males and four females were aged 56-81 years (mean 69 years). All cases had high-grade morphology in the micropapillary carcinoma and typical urothelial carcinoma. All cases with extensive MPC (n = 4) were of a high pathological stage (pT3 or pT4) and died of disease (DOD) or other causes. Eighty percent with moderate MPC (eight of 10 cases) were pT3 or pT4 and 50% DOD or are alive with disease. Eighty-four percent with focal MPC (five of six cases) were pT1 or pTa. In high-stage cases, the most invasive component was MPC. High-stage cases had an 85% risk of being advanced at presentation with micropapillary carcinoma. All pT2 or lower stage cases had micropapillary carcinoma on prior transurethral resections of bladder tumour (TURB). High-stage carcinomas had 30% and 54%, respectively, of surface MPC and urothelial carcinoma in situ, in comparison with 85% and 28% in lower stage carcinomas. Immunohistochemical staining was similarly positive in MPC and typical urothelial carcinoma with cytokeratin (CK)7, CK20, epithelial membrane antigen, carcinoembryonic antigen and cytokeratin 34betaE12. CA125 staining was seen only in MPC in 43% of cases.. Micropapillary urothelial carcinoma is a high-grade carcinoma in which the prognosis is related to the proportion and location of the MPC. Cases with moderate or extensive MPC are at high risk of being advanced at presentation. Cases with <10% MPC and surface MPC have a high chance of detection at an early stage. The morphology and immunohistochemical profile of the MPC suggest that it is a form of glandular differentiation in urothelial carcinoma.

Topics: Aged; Aged, 80 and over; CA-125 Antigen; Carcinoembryonic Antigen; Carcinoma, Papillary; Carcinoma, Transitional Cell; Female; Follow-Up Studies; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Middle Aged; Mucin-1; Survival Rate; Urinary Bladder Neoplasms

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Carcinoma, Transitional Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Middle Aged; Prostatic Neoplasms; Receptors, Androgen; Urinary Bladder Neoplasms

We report a case of mucinous adenocarcinoma of the renal pelvis associated with bladder carcinoma in situ (CIS). Transitional cell carcinoma (TCC) of the renal pelvis and CIS were also observed adjacent to the adenocarcinoma. Immunohistochemical assessment of the pelvic adenocarcinoma revealed positive expressions for mucin, epithelial membrane antigen, cytokeratin 7, cytokeratin 19 and carcinoembryonal antigen, but not vimentin or chromogranin. Based on the histopathological examinations, the adenocarcinoma of the renal pelvis in the present case may have a similar biological nature to conventional TCC and probably originated by development of pre-existing TCC of the renal pelvis.

Topics: Adenocarcinoma, Mucinous; Aged; Carcinoembryonic Antigen; Carcinoma in Situ; Carcinoma, Transitional Cell; Humans; Keratin-7; Keratins; Kidney Neoplasms; Kidney Pelvis; Male; Mucin-1; Mucins; Neoplasms, Multiple Primary; Urinary Bladder Neoplasms

To evaluate the value of urine tests based on the detection of cytokeratins 8 and 18 for the diagnosis of bladder cancer compared with urine cytology.. Samples from 112 patients before transurethral resection (group 1), 40 patients before secondary surgical treatment (group 2), 29 healthy control subjects (group 3, controls), and 10 women with acute urinary tract infection (group 4, controls) were examined with the UBC Rapid and UBC II enzyme-linked immunosorbent assay (ELISA) tests and voided urine cytology.. Of the 112 patients in group 1, 90 had transitional cell carcinoma. For the UBC Rapid, UBC ELISA, and cytology, the sensitivity and specificity was 64.4%, 46.6%, and 70.5% and 63.6%, 86.3%, and 79.5%, respectively. The cytology had the greatest accuracy (72.3%) compared with both cytokeratin tests (54.4% and 64.2%). For all three tests, sensitivity increased with tumor grade and stage. In group 2, 16 of 40 patients had residual carcinoma. The sensitivity was similar for all three tests, and the specificity of cytology was lower compared with its specificity in group 1 (47.9% versus 70.5% in group 1). In the controls with or without urinary tract infection, the specificity of cytology was greater than that of both other tests. The combination of the UBC ELISA test with cytology increased the sensitivity to 83% (specificity 68%).. Both cytokeratin tests detected patients with transitional cell carcinoma, but were inferior to voided urine cytology in test quality.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Middle Aged; Sensitivity and Specificity; Urinalysis; Urinary Bladder Neoplasms; Urinary Tract Infections; Urine

The morphology of urothelial carcinomas, particularly when poorly differentiated or in metastatic sites, is not distinctive and overlaps significantly with other poorly differentiated nonurothelial carcinomas. Currently, there is no widely used single marker or panel of markers to confirm urothelial origin. We evaluated a panel consisting of antibodies to uroplakin III (UROIII), thrombomodulin (THR), high molecular weight cytokeratin (HMWCK), and cytokeratin 20 (CK20) in a wide range of urothelial tumors. Immunohistochemistry was performed on 112 paraffin-embedded urothelial neoplasms: 14 low malignant potential, 16 low-grade noninvasive, 16 high-grade noninvasive, 36 invasive, and 25 metastatic and 5 small cell carcinomas of the urinary bladder. Tissue microarray analysis was used to examine 498 tissue cores of nonurothelial tumors and normal tissue using antibodies to UROIII, THR, and HMWCK. Overall positive staining results in all urothelial tumors are as follows: UROIII, 64 of 112 (57.1%); THR, 77 of 112 (68.8%); HMWCK, 88 of 110 (80%); and CK20, 53 of 110 (48.2%). The expression of the four markers varied with tumor grade and stage. All small cell carcinomas were negative for all markers. Variant morphologic subtypes showed similar staining as conventional urothelial carcinomas. Tissue microarray analysis showed no UROIII immunoreactivity in tissue cores of nonurothelial tumors. THR was expressed by a limited number of nonurothelial cores (10 of 37 [27%] non-small cell lung carcinomas, 2 of 36 [5.6%] lymphomas). HMWCK was expressed by 43.8% of non-small cell lung carcinomas and essentially absent in other nonurothelial tumor cores. Based on the results of the study, the expression of UROIII in a tumor is essentially diagnostic of urothelial origin; however, it is expressed in only slightly more than half of urothelial tumors. The coexpression of THR, HMWCK, and CK20 strongly suggests urothelial origin. The coexpression of two of three non-UROIII markers (THR, HMWCK, CK20) suggests urothelial origin but requires clinicopathologic correlation. The results of the study indicate a role for an antibody panel that includes UROIII, THR, HMWCK, and CK20 in the diagnosis of urothelial tumors.

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Female; Histocytological Preparation Techniques; Humans; Immunohistochemistry; Keratins; Male; Membrane Glycoproteins; Sensitivity and Specificity; Thrombomodulin; Urinary Bladder Neoplasms; Uroplakin III

There is no well-established positive immunomarker for urothelial carcinoma. We evaluated the diagnostic utility of high molecular weight cytokeratin (HMWCK) antibody clone 34betaE12 in differentiating high-grade invasive urothelial carcinoma from prostate cancer.. Formalin-fixed paraffin-embedded sections from 28 cases of high-grade invasive urothelial carcinoma (20 not otherwise specified (UC-NOS), eight with glandular differentiation) and 20 cases of poorly differentiated prostate carcinoma were immunostained with a monoclonal antibody to carcinoembryonic antigen (CEA), clone 85A12 and with HMWCK antibody clone 34betaE12 after microwave pretreatment or protease 24 predigestion. All cases of UC-NOS expressed HMWCK on 34betaE12 immunostaining after microwaving or enzyme predigestion. Immunoreactivity was intense and diffuse in all the cases after microwave pretreatment, whilst with enzyme predigestion immunoreactivity was sometimes patchy with <50% tumour cells positive in 20% of cases. In comparison with 34betaE12, 85A12 was insensitive with 15% of UC-NOS cases totally CEA-negative and <50% tumour cell immunoreactivity in 60% of cases. Rare positive cells were present in two (10%) cases of prostate cancer with monoclonal anti-CEA and 34betaE12 on microwaved sections, but all the cases were HMWCK-negative using 34betaE12 on sections pretreated by enzyme digestion.. HMWCK antibody clone 34betaE12, particularly when used with microwave heat retrieval, is a very sensitive positive marker for high-grade invasive urothelial carcinoma.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Transitional Cell; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Male; Prostatic Neoplasms; Urinary Bladder Neoplasms

Immunohistochemistry for uroplakin III (UP III), cytokeratin 7 (CK 7), and cytokeratin 20 (CK 20) using commercially available antibodies was done in normal canine urinary bladder and 72 canine urinary bladder tumors that had been fixed in formalin and embedded in paraffin. Prolonged fixation (3-28 days) did not significantly alter the immunostaining for UP III. There was moderate reduction in the intensity for CK 7 and CK 20 after 1 week of fixation. UP III was detected in superficial (umbrella) cells and some intermediate cells of the normal urinary bladder, 7 of 7 transitional cell papillomas (TCPs), 50 of 55 transitional cell carcinomas (TCCs), and 4 of 5 metastatic TCCs. Staining was typically outlined in the plasma membrane, but diffuse or focal cytoplasmic staining was also observed. Intracytoplasmic lumina were usually positive for UP III. One squamous cell carcinoma of the bladder, 4 nonepithelial bladder tumors, and 285 nonurothelial tumors from different nonurinary locations were negative for UP III. CK 7 was detected in 7 of 7 TCPs, 53 of 54 TCCs, and 5 of 5 metastatic TCCs. The staining for CK 7 was diffuse cytoplasmic. CK 20 was detected in 1 of 7 TCPs, 37 of 54 TCCs, and 1 of 5 metastatic TCCs. The staining with CK 20 was cytoplasmic and weaker than with antibodies to UP III or CK 7. There was concurrent expression of UP III, CK 7, and CK 20 in 36 of 54 TCCs. UP III is a specific and sensitive marker for canine transitional epithelial (urothelial) neoplasms, detecting 91% of TCCs. Negative results may be observed with anaplastic tumors.

Topics: Animals; Carcinoma, Transitional Cell; Dog Diseases; Dogs; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Membrane Glycoproteins; Papilloma; Tissue Fixation; Urinary Bladder Neoplasms; Uroplakin III; Urothelium

Previous investigations have demonstrated the prognostic value of disseminated cytokeratin positive cells in bone marrow of patients with breast, gastric, colon and prostate cancer. We evaluated the potential of an immunocytochemical assay, using a monoclonal antibody against cytokeratin 18 (CK 18), for the detection of disseminated tumor cells in bone marrow aspirates of patients with transitional cell carcinoma.. Bone marrow aspiration was performed preoperatively on 128 patients with transitional cell carcinoma of various stages and on 27 controls with nonmalignant disease. Cytospin preparations of mononuclear bone marrow cells were incubated with a monoclonal anti-CK 18 antibody and stained using the alkaline phosphatase anti-alkaline phosphatase technique.. Of the patients with transitional cell carcinoma 29.7% and none of the controls had a CK 18 positive bone marrow result. A significant correlation between the incidence of CK 18 positive cells in bone marrow and invasive transitional cell carcinoma (p <0.01), lymph node involvement (p <0.01), medium/high grade transitional cell carcinoma (p <0.01) and tumor progression in recurrent transitional cell carcinoma (p <0.05) was demonstrated. Furthermore, the mean number of CK 18 positive cells in bone marrow aspirates of patients with stage M+ and/or N+ disease was nearly 3 times as high as that of patients without clinically evident metastatic disease (10.4 versus 3.8 CK 18 positive cells per patient).. A significant correlation between the incidence of CK 18 positive bone marrow results in patients with transitional cell carcinoma and established risk factors could be demonstrated in our study. Further prospective followup studies should be performed to determine the prognostic value of these findings.

Topics: Aged; Biomarkers, Tumor; Biopsy, Needle; Bone Marrow; Bone Marrow Neoplasms; Carcinoma, Transitional Cell; Disease Progression; Female; Humans; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Neoplasm Staging; Prognosis; Risk Factors; Survival Rate; Urinary Bladder Neoplasms

We report a urachal adenocarcinoma metastatic to both ovaries in a 50-year-old Japanese woman. Pelvic examination and imaging studies revealed a large cystic tumor occupying the pelvis and another cystic tumor between the umbilicus and the urinary bladder. A laparotomy was performed. Histopathological examination revealed a urachal tumor that was a well-differentiated invasive mucinous adenocarcinoma; the overlying urothelium was intact. The right and left ovarian tumors were well-differentiated mucinous adenocarcinomas. The urachal and ovarian tumors were immunoreactive for cytokeratin 20 and carcinoembryonic antigen, but negative for cytokeratin 7. The patient is alive with lymph node and bone metastases 6 months postoperatively. This is the eighth reported case of an adenocarcinoma of the bladder with ovarian metastasis.

Topics: Adenocarcinoma, Mucinous; Adult; Bone Neoplasms; Carcinoembryonic Antigen; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lymphatic Metastasis; Ovarian Neoplasms; Urachus; Urinary Bladder Neoplasms

The differential expression patterns of cytokeratin 20 (CK20) and 34betaE12 antigen in low-grade papillary urothelial tumors of the bladder are discussed.. A retrospective study of 120 patients with low-grade papillary bladder tumors (45 neoplasms of low malignant potential and 75 low-grade WHO G1 carcinomas) was performed. All tumors were graded in accordance with the 1998 World Health Organization/International Society of Urological Pathology (WHO/ISUP) and 1999 WHO classifications. The mean follow-up was 76.6 months (range, 36-168 mos), considering for prognostic purposes the time to first recurrence, or relapse-free interval (RFI), and the total number of recurrent patients. Immunohistochemically, normal or abnormal CK20 and 34betaE12 antigen expression patterns were determined for each patient. CK20 (clone IT-Ks) and a high-molecular weight cytokeratin (clone 34betaE12) were the monoclonal antibodies used in the immunohistochemical study.. Seventy-seven of 120 patients (64.2%) experienced a recurrence during follow-up. In recurrence prediction, the differential expression pattern of both cytokeratins showed a high sensitivity (76.6% for CK20 and 80.5% for 34betaE12 antigen) and a high positive predictive value (85.5% for CK20 and 75.6% for 34betaE12 antigen), although specificity was higher for CK20 (76.7%) than it was for 34betaE12 antigen (53.4%). Independent of adjuvant intravesical chemotherapy, these 2 markers showed a strong statistical correlation (p < 0.001) in univariate studies with both the prediction of disease recurrences and RFI.. CK20 and 34betaE12 antigen have proved to be strong predictive markers of disease recurrences when considering different topographic expression profiles, and, in the authors' opinion, these profiles could be incorporated into follow-up clinicopathologic strategies.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Papillary; Disease Progression; Disease-Free Survival; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratin-20; Keratins; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasm Staging; Prognosis; Retrospective Studies; Survival Rate; Urinary Bladder Neoplasms

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

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

The content of urinary bladder cancer antigen (UBC), tissue polypeptide specific antigen (TPS), nuclear matrix protein 22 (NMP22), and the expression of LewisY carbohydrate antigens and of epidermal growth factor receptor (EGF-R) in bladder tumor tissues were determined. These included 14 well, 6 moderately and 11 poorly differentiated bladder cancers. Cytosol UBC and TPS were higher in the well and in the moderately differentiated bladder tumors than in the poorly differentiated bladder cancers; whereas cytosol NMP22 was higher in the poorly differentiated bladder cancers than in the well and in the moderately differentiated bladder tumors. The Lewis related carbohydrate antigens, evaluated by the reactivity of the tissues to monoclonal antibody B3, were highly expressed in poorly differentiated tumors. The EGF-R was strongly expressed in a large number of poorly differentiated bladder tumors. These data suggest that the determination of cytosol NMP22 and the immunoblotting with B3 and EGF-R antibodies might be useful to obtain more information on the differentiation of bladder tumors.

Topics: Aged; Aged, 80 and over; Carbohydrates; Cell Differentiation; Cell Membrane; Cytosol; ErbB Receptors; Female; Humans; Immunoblotting; Keratins; Lewis Blood Group Antigens; Male; Middle Aged; Nuclear Matrix-Associated Proteins; Nuclear Proteins; Urinary Bladder Neoplasms

Fine-needle aspiration biopsy (FNAB) is a technique used increasingly for the investigation of primary and metastatic cutaneous tumors. Trichoblastoma is a rare benign skin appendage tumor of hair germ origin. We report the diagnosis by FNAB of a rare giant subcutaneous tumor, trichoblastoma, from an 81-yr-old woman with a subcutaneous mass in the interscapular area of her back. The cytologic characteristics of the tumor are discussed in detail in this report. The findings have been compared with the histologic features of the tumor after surgical excision. We have characterized several distinctive cytologic features that may aid in the diagnosis of this rare neoplasm. While most reported cases have been diagnosed from surgical excisional biopsy specimens, FNAB may also be a valuable tool for the accurate diagnosis of trichoblastoma in the proper clinical context.

Topics: Aged; Biomarkers, Tumor; Biopsy, Fine-Needle; Carcinoma, Transitional Cell; Combined Modality Therapy; Humans; Immunohistochemistry; Keratin-8; Keratins; Male; Middle Aged; Penile Neoplasms; Prostate-Specific Antigen; Prostatic Neoplasms; Urinary Bladder Neoplasms

Metastatic malignancy to the penis is an uncommon clinicopathologic entity, with only 300 cases reported since 1870. Of the reported cases, 75% were secondary to genitourinary primary tumors. Priapism is the most frequent symptom, although dysuria, ulceration, and node formation have also been described. We report three cases of penile metastatic involvement from primary tumors in the urinary bladder (two cases) and prostate (one case), respectively. Fine-needle aspiration (FNA) cytology from the penile nodules was performed in each case. The smears in all cases were highly cellular, and atypical neoplastic cells were observed singly, in clusters, or in papillary formations. The cells were pleomorphic with hyperchromatic nuclei and prominent nucleoli. Immunocytochemistry was performed for keratin 8 and 18 and prostatic-specific antigen (PSA). In conclusion, although it has rarely been used as a diagnostic tool, FNA of the penis can be proved effective and safe in diagnosing a suspected secondary malignancy.

Topics: Aged; Biomarkers, Tumor; Biopsy, Fine-Needle; Carcinoma, Transitional Cell; Combined Modality Therapy; Humans; Immunohistochemistry; Keratin-7; Keratins; Male; Middle Aged; Penile Neoplasms; Prostate-Specific Antigen; Prostatic Neoplasms; Urinary Bladder Neoplasms

Urothelial carcinoma with gland-like lumina is an uncommon type of tumor, reported only occasionally in literature. Its diagnosis usually does not offer any difficulties, and its prognosis is determined by the accompanying classic transitional or squamous component. It is important though, not to misdiagnose it as a mixed transitional cell adenocarcinoma. In that respect, features such as the type of epithelium lining, the gland-like structures, as well as the type of luminal mucin have been used to make the diagnosis. Recently, an immunohistochemical panel of antibodies has proven helpful in differentiating primary and metastatic adenocarcinomas of urothelial tract from urothelial carcinoma with gland differentiation. In their series of 16 cases, Tamboli et al included only one case of transitional cell carcinoma with gland differentiation. We present two additional cases of urothelial carcinoma with gland-like lumina in two men, 60 and 79 years old, respectively. Both tumors were grade 2 of Ash-Bergkvist, and the stage was pT(1) in both cases. Immunohistochemical study with cytokeratins 7 and 20, and with c-erbB-2, was performed. Both tumors expressed cytokeratins 7 and 20; c-erbB-2 was only expressed in one, in spite of the same staging. Although some relation has been found in animals between gland-like lumina phenotype and expression of epidermal growth factor (the receptor of which is homologous to c-erbB-2), it seems that this relationship might not be constant in humans.

Topics: Aged; Biomarkers, Tumor; Carcinoma; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Middle Aged; Receptor, ErbB-2; Urinary Bladder Neoplasms; Urothelium

The urinary concentration of soluble cytokeratin 19 fragments, measured by the CYFRA 21-1 assay, may be used for the noninvasive, early detection of bladder carcinoma.. This prospective study examined urine samples from 325 patients. The authors included 152 patients who presented with hematuria or irritative voiding symptoms (Group 1), 107 patients who were under surveillance after undergoing transurethral resection of bladder carcinoma (Group 2), 46 patients with urinary tract pathology other than bladder carcinoma (Group 3), and 20 healthy participants (Group 4). The urine concentration of CYFRA 21-1 was measured by an immunoradiometric assay. The patients in Groups 1 and 2 underwent cytoscopy and urine cytopathology. Biopsies were obtained if a tumor was seen on cytoscopy or if there was a suspicion of carcinoma in situ (CIS).. The optimal cut-off concentration for the detection of primary bladder tumors, 4.9 microg/L, resulted in a sensitivity of 79.3% and a specificity of 88.6%. The optimal threshold for the detection of recurrent bladder tumors (excluding patients who had been treated with intravesical bacillus Calmette-Guerin [BCG]), 4.04 microg/L, resulted in a sensitivity of 76.2% and a specificity of 84.2%. There was no significant advantage for centrifugation of the urine samples or for determination of the creatinine concentration in the urine samples. The CYFRA 21-1 assay of urine samples provided a three-fold greater sensitivity compared with the sensitivity of cytology for detecting Grade 1 transitional cell tumors. CYFRA 21-1 detected 91.9% of Grade 3 tumors, 100% of CIS, and 92.8% of invasive bladder tumors (T2 or higher classification). The CYFRA 21-1 assay detected all tumors that had positive cytology with the exception of only one tumor. Conversely, the assay identified 71% of primary tumors and 65% of recurrent tumors that were missed by cytopathology. Urinary stones, infection, and previous intravesical BCG immunotherapy caused many false positive results.. The urinary CYFRA 21-1 assay is a useful test for the noninvasive detection of bladder carcinoma and for surveillance of patients who were not treated previously with BCG. It may be used in combination with urine cytology and bladder ultrasound. Multi-institutional trials are required to compare the accuracy as well as the cost of this combination of tests with cystoscopy.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Female; Hematuria; Humans; Immunoassay; Keratin-19; Keratins; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Prospective Studies; ROC Curve; Sensitivity and Specificity; Urinary Bladder Neoplasms

The aim of the study was to compare serum levels of cytokeratin-18 of patients with bladder cancer with those of the healthy controls, and to investigate the relation between cytokeratin level and the tumor stage.. Serum cytokeratin-18 levels of 38 patients with bladder cancer and of 25 healthy people were determined. Tumor stage was T(1) in 12 patients, T(2) in 9 patients, T(3) in 10 patients and T(4) in 7 patients. The serum cytokeratin-18 levels in these cases were analyzed with respect to the stage of the tumor.. Cytokeratin-18 level in the patient group was found to be significantly higher than that of the control group (p < 0.010) when the groups were totally compared. However, when the levels in patients with different tumor stages were compared with that of the controls, the difference was not significant in patients with stage 1 and 2 tumors (p > 0.05). Regarding the cut off value as 4.0 ng/mL, sensitivity and specificity for serum cytokeratin-18 were found to be 53% and 72% respectively. When sensitivity was calculated with respect to tumor stages, it was 8% for T(1,) 33% for T(2,) 90% for T(3) and 100% for T(4.) On the other hand, considering higher stage (T(3) and T(4)) tumors only, the sensitivity was calculated as 94%, but the sensitivity for lower stage (T(1) and T(2)) tumors was 19%.. It is clear that serum cytokeratin-18 level increases in patients with bladder cancer. However, it can only be useful as a tumor marker in the diagnosis of T(3) and higher staged tumors. This study indicated that cytokeratin-18 does not have any diagnostic value in lower stage bladder cancers.

Topics: Biomarkers, Tumor; Cluster Analysis; Control Groups; Female; Humans; Keratins; Luminescent Measurements; Male; Middle Aged; Neoplasm Staging; Sensitivity and Specificity; Urinary Bladder Neoplasms

Nephrogenic adenoma is a rare benign lesion of the urinary tract involving mainly the urinary bladder. The clinical manifestations, endoscopic signs and histopathological pattern of NA should be differentiated from a cancer, which may be a source of misinterpretation. Here we report a case of a 70-year-old man previously treated for papillary urothelial carcinoma. Six months later he developed a tumour, giving rise to a suspicion of recurrence. Histopathologically the tumour was diagnosed as nephrogenic adenoma. It is the first case of nephrogenic adenoma in Polish literature.

Topics: Adenoma; Aged; Antigens, Nuclear; Biomarkers, Tumor; Humans; Immunohistochemistry; Keratins; Male; Neoplasms, Second Primary; Nuclear Proteins; Treatment Outcome; Urinary Bladder Neoplasms; Vimentin

A case of sarcomatoid carcinoma of the bladder is reported herein. Immunohistochemical staining with human pancytokeratin antibody was negative, while vimentin staining was strongly positive, suggesting a diagnosis of sarcoma of the bladder. Further immunohistochemical analysis revealed positivity for AE1/AE3 cytokeratins, permitting a correct diagnosis of sarcomatoid carcinoma of the bladder. It can be difficult to distinguish between sarcomatoid carcinoma, undifferentiated carcinoma and sarcoma, particularly if the biopsy specimens are of small size. In rare cases, sarcomatoid tumors may express epithelial markers different from those revealed by human pancytokeratin staining.

Topics: Aged; Carcinoma; Female; Humans; Immunohistochemistry; Keratins; Sarcoma; Urinary Bladder Neoplasms

A histopathological and immunohistochemical study of a case of nephrogenic adenoma of the bladder associated to glandular cystitis is presented with a very similar immunostaining to adenomatoid tumors in other organs and probably of a mesothelial origin. Its pathogenesis seems to correspond to a metaplastic change of the bladder's urothelium through anomalous differentiation of the reserve cells faced with different irritating agents. Because of its benign characteristics, we think that treatment can be confined to endoscopic observation and conservative technique.

Topics: Adenoma; Aged; Biomarkers, Tumor; Carcinoma; Cystitis; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Male; Metaplasia; Mucin-1; Neoplasm Proteins; Urinary Bladder Neoplasms; Urography; Urothelium; Vimentin; von Willebrand Factor

Pure yolk sac tumor is the most common malignant gonadal tumor of infants and toddlers. However, the majority of extragonadal germ cell tumors in the midline are either seminomas (germinomas) or teratomas, and pure yolk sac tumors account for only a small fraction of these lesions. To date, only 1 primary urachal pure yolk sac tumor has been reported in the literature. We describe another case, occurring in a 7-month-old male infant who presented with a rapidly enlarging intra-abdominal tumor with marked engorgement of the superficial venous plexus around the umbilicus. With periodic follow-up for 3 years following surgical extirpation of the tumor and adjuvant chemotherapy, this patient is still alive without evidence of disease. Notably, the glandular elements predominating in the frozen sections resulted in the initial misdiagnosis of the tumor as a urachal adenocarcinoma, although the entirely resected specimen revealed typical histologic patterns and Schiller-Duval bodies. Immunohistochemistry showed that the tumor cells were diffusely reactive to alpha-fetoprotein, alpha(1)-antitrypsin, and cytokeratin. Tumor cells were negative for p53 protein, but revealed overexpression for MDM2 protein. Flow cytometry demonstrated a diploid DNA content with S-phase being as high as 55.36%. This case emphasizes that pure yolk sac tumor can occur primarily in the remnant of the urachus in young children.

Topics: alpha 1-Antitrypsin; alpha-Fetoproteins; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Bleomycin; Chemotherapy, Adjuvant; Cisplatin; DNA, Neoplasm; Endodermal Sinus Tumor; Etoposide; Flow Cytometry; Humans; Immunohistochemistry; Infant; Keratins; Male; Nuclear Proteins; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-mdm2; Tomography, X-Ray Computed; Treatment Outcome; Urachus; Urinary Bladder Neoplasms

The diagnosis of bladder cancer is confirmed by histological analysis of tissue biopsies. Cytology of urine samples is a noninvasive alternative. The aim of this work was to find out whether flow cytometry of urine samples is more sensitive than cytology. For this purpose we studied 115 patients suspected of having bladder cancer. Cells isolated from urine samples were analyzed by cytometry for the expression of cytokeratin and CD 45 and for DNA measurements such as: DNA index, synthesis phase fraction and proliferative index (SPF + G2/M phase). At the same time we carried out cytological analysis. All positive cases were confirmed by histology (21/115), 18 were diagnosed by flow cytometry and 16 by cytology, with a sensitivity of 85.7% and 76.1%, respectively. Two cases were found to be positive by flow cytometry, which were not confirmed by histology, while no false positives were detected by cytology. We found that both techniques gave almost identical results for the diagnosis of bladder cancer, although there were differences in non-malignant samples. In conclusion, flow cytometry is slightly more sensitive than cytology but the combination of the two techniques improves the diagnosis.

Topics: Aged; Aged, 80 and over; Cytological Techniques; DNA; Female; Flow Cytometry; Humans; Keratins; Leukocyte Common Antigens; Male; Middle Aged; Ploidies; Reproducibility of Results; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urine

The subclassification of T1 bladder tumors into T1A and T1B has an important prognostic significance and a great impact on patient management. Unfortunately, staging T1 tumors is highly subject to interpathologist variation that can be critical for patients included in randomized clinical trials. To determine the value of immunohistochemistry (IHC), such as desmin and keratin, in comparison to hematoxylin-eosin (H&E) in classifying T1 stage disease, we retrospectively examined 93 consecutive cases diagnosed at our department.. The study was conducted in two phases (H&E then IHC), each in two time periods. First H&E, and then IHC slides were reviewed independently by two experienced pathologists and discrepant cases from each phase were discussed between the two pathologists to reach a final decision.. The two methodologies (H&E and IHC) showed total agreement in 76 out of 93 cases. IHC downstaged seven cases, that is from T1B to T1A, upstaged four cases, that is from T1A to T1B, lowered the rate of imprecision and eliminated the disagreement between the two pathologists. However, IHC failed to subclassify T1 tumors in three cases. Finally, the discussion supported by the IHC was very useful in reaching the diagnosis in some cases.. IHC appears to be a useful tool in staging T1 bladder cancer, especially in difficult cases where specimen orientation and artifact could create a major hindrance in reaching an accurate diagnosis.

Topics: Desmin; Humans; Immunohistochemistry; Keratins; Neoplasm Invasiveness; Neoplasm Staging; Retrospective Studies; Urinary Bladder Neoplasms

This study was designed to characterize the expression profiles of nine bladder cancer cell lines (T24, J82, 5637, HT1376, RT4, SCaBER, TCCSUP, UMUC-3, and HT1197) using cDNA microarrays (8976 genes and expressed sequence tags). Novel targets involved in bladder cancer progression of potential clinical relevance were validated by immunohistochemistry using tissue microarrays of primary bladder tumors (n = 193 cases). Hierarchical clustering classified uroepithelial cells based on their histopathogenesis and cell cycle alterations. Keratin 10 and caveolin-1 transcripts were more abundant in tumor cells from squamous and invasive origin. Their combined expression was shown to stratify bladder tumors and define squamous differentiation. To assess the robustness of the clustering analysis, a bootstrap resampling technique was used. This grouped tumor cell lines based on their biological properties, including cell cycle and cell adhesion features. E-cadherin, zyxin, and moesin were identified as genes differentially expressed in these clusters and related to the p53, RB, and INK4A status of the cell lines. Loss of these adhesion molecules was associated with stage and grade in primary tumors (P < 0.05), and moesin expression was also associated with survival (P = 0.01). Deregulation of cell cycle and apoptotic pathways, such as mutations or altered expression of p53, pRB, and INK4A (p16), is necessary for uroepithelial transformation. However, it appears that deregulation of cell adhesion is a common event associated with tumor progression in uroepithelial neoplasms.

Topics: Biomarkers, Tumor; Cadherins; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Caveolin 1; Caveolins; Cell Differentiation; Cluster Analysis; Cytoskeletal Proteins; Gene Expression Profiling; Glycoproteins; Humans; Keratin-10; Keratins; Metalloproteins; Microfilament Proteins; Monte Carlo Method; Neoplasm Staging; Oligonucleotide Array Sequence Analysis; Retinoblastoma Protein; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Zyxin

This study was designed to detect the expression of CK-19 and CK-20 in tissue specimens and IL-6 in the sera (as a noninvasive maneuver) of bladder cancer patients. Results were correlated with the clinico-pathologic parameters, Bilharziasis and the occurrence of relapse of the carcinoma among Egyptian bladder cancer patients.. Subjects of this study were 50 cases of bladder carcinoma (19 cases were positive for Bilharziasis) as well as 20 cystitis cases as control (7 cases were positive for Bilharziasis). Messenger ribonucleic acid extracted from fresh frozen tissue specimens with bladder tumor and the control group were collected and subjected to RT-PCR to detect expression of the amplification bands of CK-19 and CK-20 (214 and 370 base pairs). In the mean time, Interleukin-6 was quantified in the sera of the patients using ELISA kit.. CK-19 and CK-20 RNAs were expressed in bladder cancer cases, but not expressed in the control group. They were significantly correlated to advanced tumor stage and grade, while CK-19 positivity, was also, correlated to tumor recurrence and tumor pathology being more in SCC than TCC. Moreover, IL-6 positivity was correlated to the occurrence of malignancy, advanced grade and pathology being more in SCC than TCC. ROC curve was utilized to choose the best cut-off for serum IL-6 (49.2 pg/mL). At the determined cut-off, the sensitivity was 66% and the specificity was 95%. Bilharziasis was found to be related to advanced stages and grades of bladder cancer.. CK-19, CK-20 and IL-6 were strongly associated with malignant phenotype of Egyptian bladder tissues, so they may be used as additional markers for assessment of bladder cancer patients.

Topics: Adult; Aged; Egypt; Genetic Markers; Humans; Interleukin-6; Intermediate Filament Proteins; Keratin-20; Keratins; Middle Aged; Schistosomiasis; Urinary Bladder Neoplasms

We developed a method for culturing normal and malignant human bladder epithelial cells for many generations.. Cells from bladder washes or surgical specimens were plated in culture with lethally irradiated cells of the LA7 rat mammary tumor line (American Type Culture Collection, Bethesda, Maryland) at a confluent density and fed regularly. After cells from surgical specimens became confluent they were diluted and subcultured with fresh irradiated LA7 cells. The growth rate was measured by cell counts and cell sorter analysis. Expression of intermediate filaments was determined by immunocytochemical testing.. All 5 normal and 3 of the 4 tumor specimens developed into long-term cell strains. A single normal strain was carried through passage 11, amounting to 37 cell doublings. Cell numbers doubled in 2 days in medium with 0.5% serum and in 5.6 days in 5% serum. Plating efficiency was almost 100% and cloning efficiency was approximately 9%. LA7 conditioned medium did not stimulate bladder cell proliferation. Two tumor strains were carried through passage 9, amounting to 20 and 27 doublings, respectively. No cell strains expressed signs of senescence at culture termination. Normal and tumor strains expressed keratins 7, 10, 11, 18 and 19, and ZO1 tight junction protein but not vimentin or keratin 14. Umbrella cells comprised the uppermost cell layer in cultures from normal bladder.. LA7 feeder cells stimulate human bladder cell proliferation for many generations in culture by a juxtacrine mechanism and promote the expression of differentiated traits.

Topics: Animals; Carcinoma, Transitional Cell; Cell Culture Techniques; Cell Division; Cells, Cultured; Coculture Techniques; Culture Media, Conditioned; Epithelial Cells; Gap Junctions; Humans; Immunohistochemistry; Keratins; Mammary Glands, Animal; Rats; Tight Junctions; Tumor Cells, Cultured; Urinary Bladder; Urinary Bladder Neoplasms

To evaluate the role of BTA stat, BTA TRAK, UBC Rapid, UBC IRMA and voided urinary cytology in the detection of bladder transitional cell carcinoma (TCC).. The study included 78 patients with TCC of the bladder (group A), 62 patients with a history of bladder TCC without tumor recurrence at the time of examination (B, control group), 20 patients with other malignancy of the urinary tract (C), 38 patients with non-malignant urinary tract diseases (D), 10 patients with urinary tract infection (E) and 10 healthy volunteers (F). Except in group F, voided urine was collected before cystoscopy or cystectomy.. The specificity and sensitivity in bladder cancer detection were 87.1 and 74.4%, respectively with BTA stat, 79.3 and 48.7%, respectively with UBC Rapid, 100 and 33.3%, respectively with cytology, 72.6 and 75.6%, respectively with BTA TRAK, 64.5 and 70.5%, respectively with UBC IRMA.. The BTA stat and BTATRAK tests are superior to UBC Rapid, UBC IRMA and urinary cytology in detection of bladder TCC. In daily practice however cytology remains the best adjunct to cystoscopy because of its high sensitivity in Tis and 100% specificity. Cystoscopy cannot be replaced by any of evaluated methods.

Topics: Adult; Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Complement Factor H; Cystoscopy; Female; Humans; Keratins; Male; Middle Aged; Prognosis; Prospective Studies; Sampling Studies; Sensitivity and Specificity; Urinary Bladder Neoplasms

Cytokeratins are intermediate filament proteins which are part of the framework of eucariotic cells. A characteristic pattern of expression of cytokeratins has been described for each type of epithelium. This pattern is maintained during the process of malignancy and metastasis. Immunohistochemical analysis of cytokeratins 7 and 20 (Ck 7 and Ck 20) has been considered particularly useful for identifying the origin of carcinomas.. A total of 122 cases of transitional cell carcinoma of urinary bladder were studied immunohistochemically with monoclonal antibodies against Ck 7 and Ck 20. The peroxidase labelled-streptavidin biotin technique was performed. Cases were considered positive when at least 1% of tumoral cells were stained.. All cases investigated expressed Ck 7 whereas Ck 20 was positive in 75% of them. Reactivity for Ck 20 was always focal. No tumor was negative for Ck 7.. Ck 7+/Ck 20+ immunophenotype is highly characteristic of transitional cell carcinoma of urinary bladder, whereas Ck 7+/Ck 20- is less frequent. Immunohistochemical determination of these cytokeratins is useful for the diagnosis when tumors are poorly differentiated and for identifying the primary site of metastatic carcinomas.

Topics: Carcinoma, Transitional Cell; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Urinary Bladder Neoplasms

Paget disease of the vulva can be mimicked by several disease entities histopathologically, but most of these entities can be clinically distinguished from vulvar Paget disease. However, vulvar Paget disease is in itself a heterogeneous group of epithelial neoplasms that can be similar both clinically and histopathologically. The subtypes of vulvar Paget disease include primary Paget disease arising from a pluripotent stem cell within the epithelium of the vulva, and secondary Paget disease of the vulva. Secondary vulvar Paget disease results from spread of an internal malignancy, most commonly from an anorectal adenocarcinoma or urothelial carcinoma of the bladder or urethra, to the vulvar epithelium. We have recently proposed that these lesions be classified as primary (of cutaneous origin) or secondary (of extracutaneous origin). These subtypes can present similarly as eczematoid skin lesions and may appear similar on routine hematoxylin and eosin-stained slides. Immunohistochemical studies can help differentiate between them. Our current study includes 17 patients with a pathologic diagnosis of vulvar Paget disease. We performed a panel of immunohistochemical stains, including cytokeratin (CK) 7 and 20, carcinoembryonic antigen (CEA), gross cystic disease fluid protein-15 (GCDFP-15), and uroplakin-III (UP-III). Of these 17 patients, 14 (80%) had primary intraepithelial cutaneous Paget disease, 13 without invasion and 1 with associated invasion. Three patients had urothelial carcinoma with spread to the vulva, manifesting as secondary vulvar Paget disease. Immunohistochemically, primary vulvar Paget disease is immunoreactive for CK 7 and GCDFP-15, but uncommonly for CK 20. Vulvar Paget disease secondary to anorectal carcinoma demonstrates CK 20 immunoreactivity but is usually nonreactive for CK 7 and consistently nonimmunoreactive for GCDFP-15. Vulvar Paget disease secondary to urothelial carcinoma is immunoreactive for CK 7 and CK 20 but nonimmunoreactive for GCDFP-15. In addition, we propose the use of a new, commercially available antibody, UP-III, which is specific for urothelium and, in our experience, is immunoreactive in secondary vulvar Paget disease of urothelial origin. The distinction between these 3 types of Paget and Paget-like lesions is essential in that the specific diagnosis has a significant influence on current treatment. The difference in surgical approach to the subtypes of vulvar Paget disease justifies classifying them into disti

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratin-7; Keratins; Membrane Glycoproteins; Paget Disease, Extramammary; Urinary Bladder Neoplasms; Uroplakin III; Urothelium; Vulvar Neoplasms

To determine whether Brenner tumors and transitional cell carcinomas (TCCs) of the ovary are urothelial in type, the immunoprofiles of 14 Brenner tumors, including three malignant examples, and eight ovarian TCCs were compared with those of Walthard nests, urothelium, 12 urinary bladder TCCs and 17 ovarian adenocarcinomas (serous, endometrioid, mucinous, and undifferentiated type). The immunohistochemical stains used included those for cytokeratins CKs 5/6, CK7, CK8, CK13, and CK20, vimentin, CA125, and the specific urothelial differentiation marker uroplakin III. CK7 and CK8 were broadly expressed in most tumors of ovary and bladder examined, while vimentin was focally present in some ovarian TCCs and adenocarcinomas. As in normal and neoplastic bladder urothelium, urothelial markers, including uroplakin III, CK13, and CK20, were detected in the epithelial nests of Brenner tumors. Brenner tumor cells also expressed uroplakins Ia and II. CA125 was observed focally in some Brenner tumors. In contrast, TCCs of the ovary and Walthard nests lacked uroplakins and were essentially negative for CK20 and CK13 but quite strongly expressed CA125. This immunophenotype closely resembled that found in ovarian adenocarcinomas. Thus, it appears that the only true urothelial-type ovarian neoplasm is the Brenner tumor, whereas ovarian TCC most likely represents a poorly differentiated adenocarcinoma with a morphologic transitional cell pattern. These results may explain the controversies as expressed in the recent literature concerning TCC of the ovary and establish its place among the ovarian adenocarcinomas of müllerian type.

Topics: Adenocarcinoma; Brenner Tumor; Carcinoma, Transitional Cell; Cell Transformation, Neoplastic; Female; Humans; Keratins; Membrane Glycoproteins; Ovarian Cysts; Ovarian Neoplasms; Urinary Bladder Neoplasms; Uroplakin III; Urothelium

Despite the large number of studies performed in solid tumors, few attempts at molecular detection of urothelial cells in blood have been made. Specifically, only uroplakin II (UP-II) and cytokeratin 20 (CK-20) have been suggested as tumor markers in the blood of bladder cancer patients. Epidermal growth factor receptor (EGFR) mRNA expression was found in the blood of patients with some types of carcinoma; nevertheless, its expression has been never investigated in the blood of patients with urothelial tumors. We used a EGFR-based reverse transcription-PCR assay for the detection of tumoral cells in the blood of 27 patients with bladder cancer, in 30 healthy donors, and in 9 patients with cystitis. EGFR expression was compared with that of known markers of circulating epithelial cells, CK-19 and CK-20, and to a urothelial-specific marker, UP-II. Analysis by reverse transcription-PCR and Southern blot hybridization showed no evidence of EGFR and UP-II mRNA expression in any of the samples used as controls. Analysis of healthy donors showed mRNA expression for CK-19 and CK-20 in 6 of 30 and in 4 of 30 samples, respectively. All patients with cystitis resulted negative for EGFR expression, whereas 3 of 9, 2 of 9, and 3 of 9 were found expressing CK-19, CK-20, and UP-II, respectively. Among blood samples from tumoral patients, 74% had EGFR mRNA and 41% had positive signals for CK-19, whereas positivity for CK-20 and UP-II was found in 15% and 37% of patients, respectively. These results seem to indicate that EGFR mRNA in the blood may be a useful tumor marker in bladder cancer patients, as well as in other patients with epithelial tumors.

Topics: Adult; Biomarkers, Tumor; Blotting, Southern; Carcinoma, Transitional Cell; Cystitis; ErbB Receptors; HeLa Cells; Humans; Intermediate Filament Proteins; Keratin-20; Keratins; Lymphatic Metastasis; Membrane Proteins; Neoplastic Cells, Circulating; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Urinary Bladder Neoplasms; Uroplakin II

We study the potential diagnostic use of urinary bladder cancer antigen, CYFRA 21-1 and NMP22*; for evaluating symptomatic patients who present with microscopic hematuria and are at risk for bladder cancer.. Urinary tumor markers were determined in 187 samples from 112 patients symptomatic of bladder cancer (group 1), and 75 with benign and other urological conditions (group 2). Immunoassays were used to measure the 3 selected biomarkers. Sensitivity and specificity were established by previously defined cut points. Biomarker results were reported as corrected and uncorrected for urinary creatinine. Urinalysis was performed in all samples.. Positive and negative predictive values were 85.5%, 80.5% and 81.1%, and 80.8%, 79.2% and 76.5% for urinary bladder cancer antigen, CYFRA 21-1 and NMP22, with the cutoffs 9.7 microg./l., 5.4 microg./l and 10.0 units per ml., respectively. These predictives values were 85.2% and 72.5%, respectively, for urinary cytology. The combination of biomarkers decreased the positive predictive values to 72.3% to 78.6% and increased negative predictive values to 84.2% to 86.1%. Urinary tract infection, inflammation and malignancy associated with other genitourinary organs were the primary cause for false-positive test results in the 3 assays evaluated.. With a single biomarker, around 80% of the positive results would have correctly identified symptomatic patients for cystoscopy. Of the negative results 75% would have correctly reduced the number of cystoscopies. Sensitivity and negative predictive values could be improved with the combination of biomarkers but with a loss of specificity and positive predictive values. Urinary tract inflammation and other genitourinary malignancies might contribute to the reduction in specificity of these tests.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Creatinine; Cystoscopy; Cytodiagnosis; Female; Humans; Keratin-19; Keratins; Male; Middle Aged; Nuclear Proteins; Predictive Value of Tests; Risk Factors; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urine

-Cytokeratin 7 (CK7) and cytokeratin 20 (CK20) are 2 types of intermediate filament protein. Expression of CK7 is seen in the majority of primary urinary bladder carcinomas. CK20 is restricted to superficial and occasional intermediate cells of the normal urothelium of the bladder. Aberrant CK20 expression has been documented in urothelial carcinoma and has proved useful as an ancillary diagnostic aid for urinary bladder tumor. Our hypothesis is that the pattern of CK7 and CK20 expression in metastatic urothelial carcinoma duplicates the expression of the same markers in the primary tumors. Therefore, immunohistochemical staining of metastatic tumors for these 2 markers may be helpful for differential diagnosis in ambiguous metastatic tumor deposits.. -To determine the concordance of CK7 and CK20 expression in primary bladder urothelial carcinoma and the matched lymph node metastasis.. -We studied 26 patients with lymph node metastases who underwent radical cystectomy and bilateral lymphadenectomy for bladder carcinoma. Immunohistochemical staining for CK7 and CK20 was performed on formalin-fixed paraffin-embedded tissues containing primary cancers and lymph node metastases.. -In all cases, there was a concordant expression of CK20 in the primary cancer and its matched lymph node metastasis. Twelve cases (46%) showed positive CK20 immunoreactivity in the primary tumor and its matched lymph node metastases, whereas 14 cases (54%) were negative for CK20 in both the primary tumor and lymph node metastasis. All cases showed positive CK7 immunoreactivity in the primary cancers and matched lymph node metastases.. -CK20 immunoreactivity is reliably observed in metastases from bladder cancer when the primary tumor expresses CK20.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Staining and Labeling; Tissue Distribution; Urinary Bladder Neoplasms

Lymphoepithelioma-like carcinoma (LELCA) of the urinary bladder is a rare variant of bladder cancer characterized by a malignant epithelial component densely infiltrated by lymphoid cells. It is characterized by indistinct cytoplasmic borders and a syncytial growth pattern. These neoplasms deserve recognition and attention, chiefly because they may be responsive to chemotherapy. We report on the clinicopathologic features of 13 cases of LELCA recorded since 1981. The chief complaint in all 13 patients was hematuria. Their ages ranged from 58 years to 82 years. All tumors were muscle invasive. A significant lymphocytic reaction was present in all of these tumors. There were three pure LELCA and six predominant LELCA with a concurrent transitional cell carcinoma (TCC). The remainder four cases had a focal LELCA component admixed with TCC. Immunohistochemistry showed LELCA to be reactive against epithelial membrane antigen and several cytokeratins (CKs; AE1/AE3, AE1, AE3, CK7, and CK8). CK20 and CD44v6 stained focally. The lymphocytic component was composed of a mixture of T and B cells intermingled with some dendritic cells and histiocytes. Latent membrane protein 1 (LMP1) immunostaining and in situ hybridization for Epstein-Barr virus were negative in all 13 cases. DNA ploidy of these tumors gave DNA histograms with diploid peaks (n=7) or non-diploid peaks (aneuploid or tetraploid; n=6). All patients with pure and 66% with predominant LELCA were alive, while all patients having focal LELCA died of disease. Our data suggest that pure and predominant LELCA of the bladder appear to be morphologically and clinically different from other bladder (undifferentiated and poorly differentiated conventional TCC) carcinomas and should be recognized as separate clinicopathological variants of TCC with heavy lymphocytic reaction relevant in patient management.

Topics: Aged; Aged, 80 and over; Aneuploidy; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; DNA, Neoplasm; Female; Humans; Image Cytometry; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Mucin-1; Neoplasms, Multiple Primary; Survival Rate; Urinary Bladder Neoplasms

We present the first report of bladder carcinoma that demonstrates a mixture of two distinct histological patterns resembling malignant lymphoma. The patient was a 79-year-old man. One of the histological patterns was a diffuse growth of monomorphic carcinoma cells, and the other was a dense lymphoplasmacytic infiltrate, obscuring the carcinoma. The tumor cells showing both patterns expressed cytokeratin and epithelial membrane antigen, but not lymphoid markers. Careful immunohistochemical evaluation should be done when diagnosing urinary bladder carcinomas resembling lymphomas (other than primary lymphomas).

Topics: Aged; Carcinoma; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Lymphoma; Male; Mucin-1; Urinary Bladder Neoplasms

We report herein a case of a collision tumor composed of high-grade urothelial carcinoma and a Gleason grade 3+4 prostate adenocarcinoma metastasizing to the same lymph node. After the patient underwent cystoprostatectomy for known urothelial carcinoma, he was incidentally discovered to have a second primary prostate tumor. Lymph node examination revealed that one node appeared to have metastatic foci from both primary tumors. The presence of 2 tumor types colliding in the same lymph node was confirmed using immunohistochemical stains, including monoclonal carcinoembryonic antigen, prostate-specific antigen, prostatic acid phosphatase, cytokeratins 7 and 20, and CD57. We also stained both primary tumors with the same panel as an internal control. Although 2 similar collision tumors have been reported in the literature in the past, neither used a battery of immunohistochemical stains to definitively distinguish one tumor from the other. Herein, we review the literature on urothelial and prostate collision tumors and some of the special stains used to distinguish them.

Topics: Acid Phosphatase; Adenocarcinoma; Aged; Carcinoma, Transitional Cell; CD57 Antigens; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Neoplasms, Second Primary; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms; Urinary Bladder Neoplasms

Topics: Aged; Carcinoma, Squamous Cell; Cystectomy; Cystoscopy; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Keratins; Urinary Bladder Neoplasms

We assessed diagnostic criteria among 38 spindle cell tumors of the urinary bladder and obtained follow-up in 36 patients. Patients comprised 28 males and 10 females aged 2.5 months to 87 years. Hematuria was the commonest presenting symptom (27 patients). After review and immunohistochemical workup, 17 patients had inflammatory pseudotumor (myofibroblastic tumor), 4 postoperative spindle cell nodule, 1 leiomyoma, 13 sarcoma (7 low-grade; 6 high-grade), and 3 carcinoma. Mean age was 38 years for pseudotumor (range 15 to 74), 65 for postoperative spindle cell nodule, 51 for sarcoma, and 76 for carcinoma. Size of pseudotumor averaged 4.4 +/- 0.7 cm (range 1.5 to 13.0), similar to sarcoma, 4.0 +/- 0.6 cm (range 0.5 to 7.0). Similar proportions of benign tumors and sarcomas had muscularis propria invasion. The criteria that best differentiated sarcoma from inflammatory pseudotumor were presence of necrosis at the tumor-detrusor muscle interface in muscle-invasive cases, and nuclear atypia. Sarcoma also had less prominent microvasculature, less variable cellularity, consistently > or =1 mitotic figure per 10 high-power fields, and predominant acute inflammation without plasma cells. p53 protein nuclear immunostaining was moderate, unlike the rare to absent staining in pseudotumors. Because all 12 sarcomas were desmin-negative, we did not call them leiomyosarcoma; they overlapped with benign tumor in epithelial, mesenchymal, and actin immunostaining. Among 12 sarcoma patients, 2 died of tumor (at 3 months). Two of four experienced tumor recurrence after partial cystectomy (2 and 26 months). No pseudotumors recurred after transurethral resection or partial cystectomy, although one patient, 5 months after transurethral resection, had histologically identical pseudotumor that the surgeon considered residual. Another patient with pseudotumor, not a candidate for tumor ablation after transurethral resection, had continued tumor growth and he died of urosepsis. In conclusion, inflammatory pseudotumor, although overlapping with sarcoma in presentation, age range, and size, does not metastasize and remains histologically distinct from low-grade sarcoma.

Topics: Actins; Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; Cystectomy; Desmin; Diagnosis, Differential; Female; Follow-Up Studies; Granuloma, Plasma Cell; Humans; Immunohistochemistry; Infant; Keratins; Male; Middle Aged; Mucin-1; Muscle, Smooth; S100 Proteins; Sarcoma; Treatment Outcome; Tumor Suppressor Protein p53; Urinary Bladder; Urinary Bladder Diseases; Urinary Bladder Neoplasms; Vimentin

ECV304 cells reported as originating from human umbilical vein endothelial cells by spontaneous transformation have been used as a model cell line for endothelia over the last decade. Recently, deoxyribonucleic acid fingerprinting revealed an identical genotype for ECV304 and T24 cells (urinary bladder carcinoma cell line). In order to resolve the apparent discrepancy between the identical genotype and the fact that ECV304 cells phenotypically show important endothelial characteristics, a comparative study was performed. Immortalized porcine brain microvascular endothelial cells/C1-2, and Madin Darby canine kidney cells were included as typical endothelial and epithelial cells, respectively. Various methods, such as confocal laser scanning microscopy. Western blot, and protein activity tests, were used to study the cell lines. ECV304 and T24 cells differ in criteria, such as growth behavior, cytoarchitecture, tight junction arrangement. transmembrane electrical resistance, and activity of gamma-glutamyltransferase. Several endothelial markers (von Willebrand factor, uptake of low-density lipoprotein, vimentin) could clearly be identified in ECV304, but not in T24 cells. Desmoglein and cytokeratin, both known as epithelial markers, were found in ECV304 as well as in T24 tells. However, differences were found for the two cell lines with respect to the type of cytokeratin: in ECV304 cells mainly cytokeratin 18 (45 kDa) is found, whereas in T24 cells cytokeratin 8 (52 kDa) is predominant. As we could demonstrate, the ECV304 cell line exposes many endothelial features which, in view of the scarcity of suitable endothelial cell lines, still make it an attractive in vitro model for endothelia.

Topics: Antigens, CD; ATP Binding Cassette Transporter, Subfamily B, Member 1; Blotting, Western; Cadherins; Cell Count; Cell Division; Cell Line; Cytoskeletal Proteins; Desmogleins; Desmoplakins; Endothelium, Vascular; gamma-Glutamyltransferase; Humans; Keratins; Kinetics; Phenotype; Platelet Endothelial Cell Adhesion Molecule-1; Time Factors; Tumor Cells, Cultured; Umbilical Veins; Urinary Bladder Neoplasms; Vimentin; von Willebrand Factor

The objective of the current study was to comparatively analyze the sensitivity and specificity of flow cytometric DNA/cytokeratin 8/18 measurements and the urinary bladder carcinoma antigen (UBC) enzyme linked immunoabsorbent assay (ELISA) test for the detection of bladder carcinoma in voided urine samples.. Eighty-one fresh urine voided samples, preserved frozen for a maximum period of 3 months, belonging to patients with an active bladder carcinoma (n = 37), patients who were free of disease as confirmed by cystoscopy (n = 19), patients receiving intravesical therapy (n = 17), and individuals with other benign and malignant conditions (n = 8), were collected. Flow cytometry measurements of thawed samples were based on the detection of cytokeratin (CK) 8+ and CK18+ cells using the 3F3 and 6D7 monoclonal antibodies alone or in combination with the measurement of cell DNA contents, after propidium iodide staining. Urinary bladder carcinoma antigen test was measured by ELISA.. Patients were grouped according to the presence (n = 44) or absence (n = 29) of bladder carcinoma as confirmed by cystoscopy, and taking cutoffs of 9.7 microg/L for UBC-ELISA, 75% for the percentage of 3F3 (+) and 6D7 (+) cells, and 10.6% for the proportion of hyperdiplod cells that suggested a specificity of 83%, the individual sensitivity obtained for each parameter was 77%, 5%, 9%, and 77%, respectively. The presence of DNA aneuploid populations showed a relatively low sensitivity (36%) although it was the most specific parameter (93%). Combining UBC antigen test with the proportion of cells showing DNA content higher than 2n increased to 89% the sensitivity of the UBC antigen alone. However, false-positive results for both techniques were found in individuals with urologic diseases other than bladder carcinoma and in patients receiving intravesical therapy.. The authors' results suggest that the combined use of the UBC antigen test and DNA/cytokeratin flow cytometry double stainings for the analysis of freshly obtained urine voided samples, cryopreserved to assure cellular integrity, is of great clinical utility for the detection of tumor recurrence in patients with bladder carcinoma.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; DNA, Neoplasm; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Flow Cytometry; Humans; Keratins; Middle Aged; Reagent Kits, Diagnostic; Sensitivity and Specificity; Urinary Bladder Neoplasms

Cross-section studies have shown the diagnostic characteristics of certain urinary tumor markers for the detection of bladder carcinoma. However, the role of serial urinary tumor markers in the monitoring of patients with bladder carcinoma in daily clinical surveillance has not been completely defined yet.. The study comprised 1185 urine samples belonging to 232 patients with a previous bladder carcinoma: 106 patients under follow-up (Group 1) and 126 bladder carcinoma patients receiving intravesic instillations (Group 2). Patients were monitored with urinary tumor markers during a one-year follow-up period. Urine samples were collected before cystoscopies and in the intercystoscopic periods for patients in Group 1 and before intravesic instillations for patients Group 2. Urinary bladder carcinoma antigen (UBC), CYFRA 21-1 and nuclear matrix proteins (NMP22) were measured by immunoassays.. Monitoring of the disease with urinary tumor markers could detect recurrence sooner than scheduled cystoscopies in 27 patients (87%) for UBC, 27 patients (87%) for CYFRA 21-1, and 26 patients (84%) for NMP22 out of 31 Group 1 patients who recurred; and in 16 patients (67%) for UBC, 17 patients (71%) for cytokeratin fragments (CYFRA) 21-1, and 13 patients (54%) for NMP22 out of 24 Group 2 patients who recurred. The most relevant finding was that persistence of negative urinary markers during follow-up was largely indicative of disease free status in 65 of 75 (87%) patients of Group 1 and 31 of 102 (30%) cases of Group 2. Although false positive results were present, they were mainly associated with sporadic urinary tract infections in 10 of 75 (13%) cases of Group 1 and in 36 of 102 (35%) patients of Group 2; and with urine samples collected in the first two months at the beginning of intravesic therapy in 35 of 102 patients (34%) in Group 2.. Monitoring of bladder carcinoma patients with serial urinary tumor markers could anticipate detection of recurrence. Persistent negative results might postpone and reduce the number of cystoscopies. Once the limitations leading to false positive results are controlled by urinalysis and by starting sample collection when basal levels are reached in patients with intravesic therapy, urinary tumor markers might eventually individualize the intervals between cystoscopies in the surveillance of patients with bladder carcinoma.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Cystoscopy; Female; Humans; Keratin-19; Keratins; Male; Nuclear Proteins; Urinary Bladder Neoplasms

Primary adenocarcinoma of the seminal vesicles is an extremely rare neoplasm. Because prompt diagnosis and treatment are associated with improved long-term survival, accurate recognition of this neoplasm is important, particularly when evaluating limited biopsy material. Immunohistochemistry can be used to rule out neoplasms that commonly invade the seminal vesicles, such as prostatic adenocarcinoma. Previous reports have shown that seminal vesicle adenocarcinoma (SVCA) is negative for prostate-specific antigen (PSA) and prostate-specific acid phosphatase (PAP); however, little else is known of its immunophenotype. Consequently, we evaluated the utility of cancer antigen 125 (CA-125) and cytokeratin (CK) subsets 7 and 20 for distinguishing SVCA from other neoplasms that enter the differential diagnosis. Four cases of SVCA-three cases of bladder adenocarcinoma and a rare case of adenocarcinoma arising in a mullerian duct cyst-were immunostained for CA-125, CK7, and CK20. Three of four cases of SVCA were CA-125 positive and CK7 positive. All four cases were CK20 negative. All bladder adenocarcinomas and the mullerian duct cyst adenocarcinoma were CK7 positive and negative for CA-125 and CK20. In addition, CA-125 immunostaining was performed in neoplasms that commonly invade the seminal vesicles, including prostatic adenocarcinoma (n = 40), bladder transitional cell carcinoma (n = 32), and rectal adenocarcinoma (n = 10), and all were negative for this antigen. In conclusion, the present study has shown that the CK7-positive, CK20-negative, CA-125-positive, PSA/PAP-negative immunophenotype of papillary SVCA is unique and can be used in conjunction with histomorphology to distinguish it from other tumors that enter the differential diagnosis, including prostatic adenocarcinoma (CA-125 negative, PSA/PAP positive), bladder transitional cell carcinoma (CK20 positive, CA-125 negative), rectal adenocarcinoma (CA-125 negative, CK7 negative, CK20 positive), bladder adenocarcinoma (CA-125 negative), and adenocarcinoma arising in a mullerian duct cyst (CA-125 negative).

Topics: Adenocarcinoma; Biomarkers, Tumor; CA-125 Antigen; Carcinoma, Transitional Cell; Cysts; Diagnosis, Differential; Genital Neoplasms, Male; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Mullerian Ducts; Prostatic Neoplasms; Rectal Neoplasms; Seminal Vesicles; Urinary Bladder Neoplasms

Paraganglioma of the urinary bladder is rarely encountered and its biologic behavior is uncertain. The authors sought to determine the prognostic factors that would predict patient outcome.. The Mayo Clinic experience over 53 years with paraganglioma of the bladder was reviewed. All histologic slides from 16 patients were reviewed by the authors. Eight cases were examined immunohistochemically with cytokeratin (AE1/3, cytokeratin 7, and cytokeratin 20), vimentin, S-100 protein, neuroendocrine markers (chromogranin, synaptophysin, and neuron specific enolase), p53 protein, and MIB-1. DNA ploidy was determined by digital image analysis in formalin fixed, paraffin embedded tissue. The mean follow-up was 6.3 years (range, 0.4-16.4 years).. Paraganglioma usually occurred in young adult women (mean age, 45 years; range, 16-74 years). The male-to-female ratio was 1 to 3. The common symptoms and signs were hypertension and hematuria. The tumors were usually located intramurally in the lateral and posterior wall of the bladder and were multifocal in 3 cases (18%). Seven patients were treated by transurethral resection, eight by partial cystectomy, and one by radical cystectomy. T classification was T1 (1 patient), T2 (9 patients), T3 (2 patients), and T4b (4 patients). At the time of diagnosis, one patient had distant metastasis and one had regional lymph node metastasis. One patient developed metastasis 1 year after diagnosis and died of the disease 1.5 years later. None of the patients with T1 or T2 tumors had recurrence or tumor progression. All tumors were aneuploid. The mean MIB-1 labeling index was 1.5% (range, 0.03-7.0%). The tumor cells displayed immunoreactivity for S-100 protein and neuroendocrine markers and were negative for p53 (except 1 case) and cytokeratin.. Paraganglioma of the urinary bladder occurs mostly in young adult women. Patients with tumor of advanced classification (>/=T3) are at risk of recurrence, metastasis, and dying of the disease, whereas patients in this study with T1 or T2 disease had favorable outcomes after complete tumor resection.

Topics: Adolescent; Adult; Aged; Biomarkers, Tumor; DNA, Neoplasm; Female; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Male; Middle Aged; Nerve Tissue Proteins; Paraganglioma; Pheochromocytoma; Ploidies; Prognosis; Retrospective Studies; S100 Proteins; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms; Vimentin

To compare the diagnostic value of two enzyme-linked immunosorbent assay (ELISA) tests, the nuclear matrix protein 22 (NMP22) test and a newly developed urinary bladder cancer (UBC) test, in patients having symptoms suggestive of urothelial cell carcinoma (UCC) and patients under follow-up after transurethral resection (TUR).. Two hundred forty patients with a mean age of 65.8 years (range 22 to 92) were included in this retrospective study. The tests were performed on previously frozen urine samples. Eighty-one patients had symptoms suggestive of bladder cancer and 159 patients were being followed up after complete TUR of UCC. Voided urine was evaluated by the NMP22 test and the monoclonal UBC-ELISA test, which traces cytokeratins 8 and 18. All patients underwent subsequent cystoscopy and biopsy evaluation of any suspicious lesion. The cutoff levels for bladder cancer positivity were 10 U/mL for the NMP22 test and 12 microg/L for the UBC test.. In the 54 patients with histologically proved UCC, the NMP22 test had a sensitivity of 55.5% and the UBC test a sensitivity of 64.8%. According to the histologic stages, the sensitivity of NMP22 was 51.7% in pTa tumors, 46.1% in pT1, and 70% in pT2 or higher tumors; the sensitivity of UBC was 62.1% in pTa, 53.8% in pT1, and 80% in pT2 or higher tumors. For histologic grades 1 to 3, the sensitivity was 50%, 50%, and 68.7% for NMP22 and 66.6%, 60%, and 68.7% for UBC, respectively. The specificity was 79% and 92% for NMP22 and UBC, respectively.. The monoclonal UBC-ELISA test is superior to the NMP22 test in both sensitivity and specificity. Nevertheless, neither test can replace cystoscopy.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Cystoscopy; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Middle Aged; Nuclear Proteins; Predictive Value of Tests; Retrospective Studies; Sensitivity and Specificity; Urinary Bladder Neoplasms

We studied the expression of cytokeratin (CK)-7 and CK-20 in prostate adenocarcinoma and urothelial carcinoma and evaluated their usefulness for distinguishing high-grade forms of these tumors. We examined prostate adenocarcinoma in 59 radical prostatectomy specimens and in 10 autopsy specimens showing metastatic disease, and urothelial carcinoma of the bladder in 28 cystectomy specimens. Immunohistochemical staining for CK-7, CK-20, and prostate-specific antigen (PSA) was performed on paraffin sections. For prostate adenocarcinoma, 5 cases had only CK-7 positivity, 5 had only CK-20 focal positivity, 1 stained for both markers, and 48 were negative for both. PSA was positive in all but 1 poorly differentiated prostatic carcinoma. In the autopsy cases, PSA was expressed in the prostate and the metastatic tumors in most cases; few cases were focally positive for CK-7 or CK-20, but none was positive for both markers. For the urothelial tumors, CK-7 was the sole positive marker in 6 cases, and CK-20 in 1 case; 17 cases were positive for both, and 4 were negative for both. All urothelial carcinomas were PSA negative. Although PSA is useful for differentiating prostatic from urothelial carcinoma, CK-7 and CK-20 are helpful when both are positive, supporting the diagnosis of urothelial carcinoma. However, if only 1 marker is positive or both are negative, these markers have limited usefulness for distinguishing these carcinomas.

Topics: Adenocarcinoma; Carcinoma, Transitional Cell; Humans; Immunoenzyme Techniques; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Prostate-Specific Antigen; Prostatic Neoplasms; Urinary Bladder Neoplasms

The development of urinary tumor markers such as UBC, CYFRA 21-1 and NMP22 appeared to be non invasive alternative methods for the detection of bladder cancer. We compared the individual and combined sensitivity of the urinary tumor markers in the detection of bladder cancer, contrasting them with the conventional diagnostic procedures.. 237 voided urines from subjects under risk for bladder cancer were collected immediately before the endoscopic examinations: 44 patients under suspicion of a primary bladder tumor and 193 patients under follow-up of a previous bladder cancer were included. UBC and NMP22 were measured by enzyme-immunoabsorbent-assays and CYFRA 21-1 by an electro-chemiluminescense-immunoassay.. Taking the cutoffs of 9.7 micrograms/l for UBC, 5.4 ng/ml for CYFRA 21-1 and 10.0 U/ml for NMP22 sensitivities were 70%, 69% and 67% for UBC, CYFRA 21-1 and NMP22 at specificities of 95%, 94% y 80%, respectively. All tumor markers showed higher sensitivities than urinary cytology (7%), microhematuria (62%) and gross hematuria (10%) at specificities of 99%, 78% and 99%, respectively. The combinations of NMP22 plus CYFRA 21-1 reached the highest sensitivity (79%), slightly lower than simultaneously measuring the three tumor markers (80%).. The sensitivities of the urinary markers UBC, CYFRA 21-1 and NMP22 appeared to be high enough so as to substitute urinary cytology. The diagnostic similarity between cytokeratins individually and in each type of patients might not recommend their simultaneous determination. The combined measurement of NMP22 and one cytokeratin marker (CYFRA 21-1 or UBC) appeared to be the most recommended.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antigens, Nuclear; Biomarkers; Biomarkers, Tumor; Female; Humans; Keratin-19; Keratins; Male; Middle Aged; Nuclear Proteins; Prospective Studies; Sensitivity and Specificity; Urinary Bladder Neoplasms

In 1999, the World Health Organization (WHO) published a new classification of papillary urothelial tumors of the urinary bladder. Intended to represent a reproducible, easy-to-use classification system that better separates patients with true malignancies (bladder cancer) from those patients who are at an increased risk for developing bladder cancer, problems in the differential diagnosis of various lesions remained. Probably the most critical distinction is between papillomas, papillary urothelial neoplasms of low malignant potential (lmp), and grade I papillary carcinomas. Conversely, problems in the distinction between reactive atypia, atypia of unknown significance, and dysplasia, as well as the distinction of dysplasia from carcinoma in situ (CIS), are unresolved. Whether urothelial basal cell status assessment on hematoxylin and eosin-stained slides completed by cytokeratin immunohistochemistry with anticytokeratin clone 34betaE12 may help to improve some of the previously mentioned diagnostic dilemmas was investigated. Basal cell status assessment was helpful in the differentiation between dysplasia and CIS. In dysplasia, CK IHC showed a predominantly basal labeling pattern, whereas in CIS, labeling of all urothelial layers was seen. Basal cell status assessment could separate 2 groups of pTa GIb papillary carcinoma. Group 1 with a continuous basal CK labeling and a low MIB-1 labeling index (LI) was compared with group 2, with a diffuse labeling pattern and a significantly higher MIB-1 LI. Whether group 1 carcinomas should better be assigned to the group of papillary urothelial neoplasms of lmp is discussed.

Topics: Antigens, Nuclear; Biopsy; Carcinoma, Papillary; Diagnosis, Differential; Humans; Hyperplasia; Immunohistochemistry; Keratins; Ki-67 Antigen; Nuclear Proteins; Papilloma; Urinary Bladder Neoplasms; Urothelium; World Health Organization

To compare the diagnostic value of two rapid tests, the bladder tumor antigen (BTA stat) test and the newly developed urinary bladder cancer (UBC) Rapid test, in patients having symptoms suggestive of urothelial cell carcinoma (UCC) and patients being followed up after transurethral resection.. One hundred eighty patients with a mean age of 65.8 years (range 22 to 92) were included in the present study. The tests were performed on voided urine samples. Fifty-seven patients had symptoms suggestive of UCC and 123 patients were being followed up after complete transurethral resection of UCC. The voided urine was evaluated by the BTA stat and UBC Rapid test, which detects cytokeratins 8 and 18. All patients underwent subsequent cystoscopic evaluation and biopsy of any suspicious lesion.. In 53 patients with histologically proved UCC, the BTA stat had a sensitivity of 52.8% and the UBC Rapid test of 66%. According to the histologic stage, the sensitivity of the BTA stat was 42.8% in pTa tumors, 61.5% in pT1, and 70% in pT2 or higher tumors. The sensitivity of the UBC test was 60.7% in pTa, 69. 2% in pT1, and 80% in pT2 or higher tumors. For histologic grades 1 to 3, the sensitivity was 38.8%, 52.6%, and 68.7% for the BTA stat and 44.4%, 78.9%, and 75% for the UBC Rapid test, respectively. The specificity was 70% and 90% for the BTA stat and UBC Rapid test, respectively.. The UBC Rapid test was superior to the BTA stat in both sensitivity and specificity. Both assays are simple office procedures and require no special knowledge. However, they cannot replace, but only lower, the number of cystoscopies during the follow-up of patients with previous UCC of the bladder.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Chromatography; Complement Factor H; Cystoscopy; Evaluation Studies as Topic; Follow-Up Studies; Humans; Keratins; Middle Aged; Sensitivity and Specificity; Urinary Bladder Neoplasms

The purpose of the present study was to assess a new quantitative urinary tumor marker for transitional cell carcinoma of the urinary bladder (TCC), measuring fragments of cytokeratin 8 and 18 in the urine (UBC). Urine samples of 355 individuals (77 healthy volunteers, 111 patients with benign urologic disorders, 167 patients with histologically proven bladder cancer) were examined for the presence of UBC antigen. Samples of all patients were obtained prior to therapy. Compared to healthy volunteers or patients with benign urologic disease, patients with TCC had significantly higher median urinary levels of UBC antigen (0 vs. 4.18 vs. 7.46 microg/g creatinine; p<0.001, and p<0.01, respectively). UBC antigen levels were positively correlated with tumor grade and stage. Patients with invasive TCC had significantly higher levels of UBC antigen than patients with superficial TCC (p<0.001). Elevated levels of UBC antigen were also found in patients with benign urologic disorders (median: 4.18 microg/g creatinine vs. 7.46 microg/g creatinine in cancer patients). Using a cutoff of 14.06 microg/g creatinine (corresponding to 95% specificity in the group of healthy individuals), sensitivity of UBC antigen ranged between 21.6% (pTa) and 75% (pT4). Overall specificity was 76.6%. Based on our data we conclude that the UBC antigen test in its current format is not clinically useful for detection of bladder cancer.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Case-Control Studies; Female; Humans; Keratins; Male; Neoplasm Invasiveness; Quality Control; Sensitivity and Specificity; Urinary Bladder Neoplasms

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

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

A case of carcinosarcoma of the urinary bladder in a 2-year-old girl is reported. The tumor, measuring 34 x 20 x 18 mm, was located in the peri-trigone area of the urinary bladder with polypoid features. Histologic examination revealed transitional cell carcinoma at the tumor surface with downward invasion. Concurrently, a sarcomatous area was found beneath the carcinoma, with these two different malignant components sharing on apparent transition without distinct boundaries. Sarcomatous components included immature round cells focally showing rhabdoid features. No rhabdomyomatous component was observed. Immunohistochemistry disclosed vimentin and cytokeratin-double positive cells at the transposition between carcinoma and sarcomatous components. In addition, ultrastructural analysis revealed that the epithelial cells had a distinct junctional complex, and the sarcomatous cells occasionally had a meshwork of cytoplasmic intermediate filaments, indicating bidirectional cytodifferentiation to epithelial and mesenchymal elements. The extremely young age at which this case of carcinosarcoma occurred suggests that the tumor may be of mesodermal stem cell origin.

Topics: Carcinosarcoma; Child, Preschool; Female; Humans; Immunohistochemistry; Keratins; Mesoderm; Microscopy, Electron; Mucin-1; Phosphopyruvate Hydratase; Rhabdoid Tumor; S100 Proteins; Stem Cells; Urinary Bladder Neoplasms; Vimentin

A total of 238 cases of bladder carcinoma stages Ta, Tis, T1 were submitted prospectively to multiparameter flow cytometry and immunohistochemical study in order to determine the biological aggressiveness of the tumour. DNA index (DI), S-phase fraction (SPF) obtained by bivariate cytokeratin 7/DNA analyses, and the immunohistochemical evaluation of p53 and MIB-1 were studied in relation to the traditional prognostic factors in bladder cancer (stage and grade). the variance analysis results showed that DNA aneuploidy was significantly associated with high stage (p = 0.0001), high grade (p = 0.0001), high SPF value > or = 5.5% (p = 0.0001), MIB-1 positivity > or = 31% (p = 0.0001) and high expression of p53 (staining involving > 50% of cells, p = 0.0001). Even if there was no statistical significance the hypotetraploid class (1.70 < DI < 1.89) showed poor prognostic biomarkers more frequently than the other aneuploid classes. Out of 238 cases, 101 were also submitted to flow cytometric measurement of MIB-1 (fMIB-1) to study the correlation between cell proliferation and DNA content. Data obtained from fresh, 3:1 methanol/acetone fixed samples were compared with values obtained from both cell cycle analysis methods and routine application of the MIB-1 immunostaining in histological sections. fMIB-1 values were positively correlated with SPF values (r = 0.801, p < 0.01) and S+G2M fraction (percentage of cells in S and in G2M phases) (r = 0.763, p < 0.01) but no correlation with paraffin sections was found. A fMIB-1 value > 7% was strongly associated with aneuploidy (p = 0.0001). The determination of DNA content coupled with the study of the epithelial (cytokeratin 7) and proliferative (MIB-1) markers could be useful in providing important information on the biological behaviour of superficial bladder tumours.

Topics: Adult; Aged; Aged, 80 and over; Aneuploidy; Antigens, Nuclear; Biomarkers, Tumor; Carcinoma, Transitional Cell; Cell Cycle; Cell Division; Diploidy; Disease Progression; DNA, Neoplasm; Female; Flow Cytometry; Fluorescent Antibody Technique, Indirect; Humans; Keratins; Ki-67 Antigen; Male; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Nuclear Proteins; Prospective Studies; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

In urothelial low-grade carcinomas of the bladder stage pT1, prognosis in general is good. In a subset of these tumors infiltrating beyond the lamina muscularis mucosae, prognosis clearly worsens. Unfortunately, evaluation of the lamina muscularis mucosae often is very difficult or even impossible because of its incomplete extension. In an immunohistochemical study on 131 pTa and pT1 urothelial tumors without provable lamina muscularis mucosae, we evaluated the proliferative activity with the monoclonal antibody MIB-1 and the expression pattern of cytokeratins of high molecular weight with the monoclonal antibody 34betaE12. The highest proliferative indices were found in tumors with a diffuse expression pattern of MIB-1 and 34betaE12. A preliminary analysis of follow-up data showed that 70.6% of the pT1 GIb-GIa tumors that recurred showed a diffuse expression pattern for both markers. Whether these patients are candidates for a doser follow-up or even for a more radical therapy has to be subject to further follow-up studies.

Topics: Antigens, Nuclear; Carcinoma; Cell Division; DNA-Binding Proteins; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Nuclear Proteins; Prognosis; Urinary Bladder Neoplasms

Although rare, renal cell carcinoma (RCC) can metastasize to the bladder. When this occurs, it might complicate diagnosis. Morphologically, RCC can be confused with transitional cell carcinomas (TCCs), especially those exhibiting clear cell features, and also with other bladder tumors, such as paragangliomas and metastatic melanomas. We report seven cases of RCC metastatic to the bladder that occurred in 6 men and 1 woman who were 35 to 69 years old. The most common presenting symptom was the reappearance of hematuria, which developed from 2 to 131 months (mean, 41.3 mo) after the removal of the primary RCC. In all of the patients, the metastatic RCC involved multiple organs; no case had an isolated metastasis to the bladder. The prognosis was poor, and five patients died of disease between 4 and 24 months (mean, 12.8 mo) after diagnosis of the metastasis to the bladder. The remaining two patients were lost to follow-up. All of the tumors were conventional clear or "granular" cell RCCs, with nuclear grades of 2 or 3. In five patients, metastases were confined to the lamina propria, but in two patients, tumors involved the muscularis propria as well. A comparative immunohistochemical study showed that metastatic RCCs were positive for CAM5.2, vimentin, and Leu-M1, and negative for cytokeratin 20, cytokeratin 7, 34betaE12, carcinoembryonic antigen, S-100 protein, HMB45, and chromogranin. Classic and clear cell TCCs were positive for all of the cytokeratins and carcinoembryonic antigen and negative for vimentin. Paragangliomas were positive for chromogranin and showed scattered positivity for the S-100 protein in the sustentacular cells. Metastatic melanomas were positive for S-100 protein and HMB45. The histologic appearance of RCC, particularly the delicate fibrovascular stroma with abundant sinusoidal vessels, is a feature that can be used to recognize the tumor. When there is difficulty diagnosing metastatic RCC, TCC, or other tumors in the bladder, the immunohistochemical findings can assist in the differential diagnosis.

Topics: Adult; Aged; Antigens, Neoplasm; Carcinoembryonic Antigen; Carcinoma, Renal Cell; Carcinoma, Transitional Cell; Chromogranins; Diagnosis, Differential; Female; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Kidney Neoplasms; Lewis X Antigen; Male; Melanoma; Melanoma-Specific Antigens; Middle Aged; Neoplasm Proteins; Paraganglioma; S100 Proteins; Urinary Bladder Neoplasms; Vimentin

Small cell (neuroendocrine) carcinoma of the urinary bladder is clinically more aggressive than urothelial (transitional cell) carcinoma. We have investigated the immunohistochemical markers most useful in diagnosing small cell carcinoma in bladder.. We evaluated the expression of chromogranin A, CD44 variant 6 (CD44v6), cytokeratin (CAM 5.2), gamma-enolase, synaptophysin, and CD45 in 46 small cell carcinomas of the bladder. Small cell and urothelial carcinoma were mixed in 21 (46%) cases. The two immunohistochemical markers with best ability to discriminate between small cell and urothelial carcinoma were chromogranin A and CD44v6. Chromogranin A had 97% specificity for small cell carcinoma, staining 65% of cases with 2+/3+ mean intensity; only one case (5%) of urothelial carcinoma was weakly (1+/3+) positive. CD44v6 was 80% specific for urothelial carcinoma, with immunoreactivity in 60% of cases, compared with 7% of small cell carcinoma cases. In cases positive for CD44v6, the mean percentage of reactive urothelial carcinoma cells was 75% (range 10-100%), greater than the 12% of cells in three cases of small cell carcinoma (P = 0.31); further, the pattern of immunoreactivity was membranous vs. focal cytoplasmic, respectively. All small cell carcinomas stained with one of the three neuroendocrine markers tested; 76% of cases were reactive for synaptophysin and 93% for gamma-enolase, with specificities of 86% and 73% in comparison to urothelial carcinoma. gamma-enolase staining of small cell carcinoma was more intense (P = 0.01) than for urothelial carcinoma. Cytokeratin CAM 5.2 stained a mean 47% of cells in small cell carcinoma, always in a punctate perinuclear pattern, and 75% in urothelial carcinoma, in a membranous pattern.. CD44v6, chromogranin A, and possibly gamma-enolase and cytokeratin (CAM 5.2) help differentiate small cell carcinoma from urothelial carcinoma.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Small Cell; Carcinoma, Transitional Cell; Chromogranins; Diagnosis, Differential; Female; Glycoproteins; Humans; Hyaluronan Receptors; Immunoenzyme Techniques; Keratins; Male; Middle Aged; Phosphopyruvate Hydratase; Urinary Bladder Neoplasms

Nephrogenic Adenoma (NA) is a rare lesion of the urinary tract, considered a metaplastic response to chronic inflammation, trauma or immunosuppression.. We report two cases of NA arising in the urinary bladder of patients with previous history of recurrent urinary tract infections due to neuropsychiatric disease. Pathological examination of the lesions, resected by transurethral (TUR) management, revealed a papillary proliferation of tubules and cysts lined by cuboidal to low-columnar cells without atypia. Immunohistochemistry showed positivity for Cam 5.2, CK7 and EMA. MIB 1 count demonstrated a positivity in 12/200 cells in case 1 and < 2/200 in case 2. No expression of nuclear p53 was evident.. NA is a benign unusual neoplasm which might be misdiagnosed as clear cell adenocarcinoma of the bladder or prostatic adenocarcinoma. Its recognition is important because it is a benign lesion cured by a conservative resection and no additional therapy is generally required.

Topics: Adenocarcinoma; Adenocarcinoma, Clear Cell; Adenoma; Antigens, Nuclear; Biomarkers, Tumor; Calbindin 2; Carcinoma in Situ; Carcinoma, Transitional Cell; Diagnosis, Differential; Epithelial Cells; Female; Humans; Immunophenotyping; Keratins; Ki-67 Antigen; Male; Metaplasia; Middle Aged; Mucin-1; Neoplasm Proteins; Neoplasms, Multiple Primary; Nuclear Proteins; Prostatic Neoplasms; Protein Isoforms; S100 Calcium Binding Protein G; Urinary Bladder Neoplasms

Immunohistochemistry for keratin has enhanced our ability to detect micrometastases in certain cancer patients with negative lymph nodes by routine histologic examination of H&E-stained sections. However, there is no information about micrometastasis of bladder cancer. We performed immunohistochemistry for keratins on 159 pelvic lymph nodes, which were negative for metastatic tumors on routine H&E-stained sections, from 19 patients with high-grade muscle invasive urothelial bladder cancer. In 1 man, 1 lymph node contained a keratin-positive micrometastasis that was not present on the original H&E-stained slide. However, the metastasis was seen readily on a new H&E-stained section prepared from the paraffin block adjacent to the keratin-stained section. Immunohistochemical analysis for keratins revealed no additional case of micrometastasis of urothelial carcinoma of the bladder. The perinodal fibroadipose tissue of a lymph node from a woman contained a few keratin-positive benign glands of endosalpingiosis. A thorough examination of the H&E-stained sections is the best method for detecting lymph node metastases of urothelial carcinoma from the bladder. There is a potential risk for misdiagnosis of metastases by using immunohistochemistry or polymerase chain reactions for keratins because of the occasional presence of benign epithelial cells in pelvic lymph nodes and associated connective tissue.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Transitional Cell; Female; Fluorescent Antibody Technique, Indirect; Humans; Keratins; Lymph Nodes; Lymphatic Metastasis; Male; Middle Aged; Pelvis; Urinary Bladder Neoplasms

We compare the individual and combined sensitivity of urinary CYFRA 21-1, urinary bladder cancer antigen, tissue polypeptide antigen and NMP22 to detect bladder cancer, evaluate the false-positive rates for different pathological conditions, and assess differential sensitivity regarding histological and clinical characteristics of disease.. A total of 267 subjects entered the study. Sensitivities of the tests were evaluated in 111 patients with active bladder cancer and 76 with no evidence of disease. False-positive rates were evaluated in 80 symptomatic and asymptomatic controls, including patients with benign urological conditions and nonbladder malignancies, and healthy subjects. CYFRA 21-1 was determined by electrochemoluminescent immunoassay in the Elecsys 2010, urinary bladder cancer antigen was quantified by enzyme linked immunosorbent assay (IDL Biotech), tissue polypeptide antigen was measured by the Prolifigen TPA-IRMA and NMP22 was assayed by enzyme linked immunosorbent assay (Matritech). Cutoffs were obtained by the 95% percentile in patients with no evidence of disease, which gave a 95% specificity for all biomarkers. Differences in sensitivity of urinary biomarkers regarding stage, grade, tumor size, pattern of growth, focality and recurrence were evaluated.. At a specificity of 95% cutoffs were 5.4 ng./ml. for CYFRA 21-1, 15.5 microg./l. for urinary bladder cancer antigen, 760.8 U./l. for tissue polypeptide antigen and 14.6 U./ml. for NMP22. Using these cutoffs sensitivities were 75.7% for NMP22, 83.8% for CYFRA 21-1, 73.9% for urinary bladder cancer antigen quantitative and 80.2% for tissue polypeptide antigen. The additional determination of cytokeratins increased the sensitivity of NMP22. Cytokeratins did not appear to be specific for bladder cancer, and false-positives rates were between 20% for urinary bladder cancer antigen and 36% for tissue polypeptide antigen for benign urological conditions, and between 40% and 52%, respectively, for nonbladder malignancies. NMP22 showed lower false-positives rates, mainly for benign diseases. Urinary tumor markers appeared to be associated with some of the most relevant histological and clinical parameters of bladder cancer.. Our preliminary evaluation showed the tests to be potential noninvasive adjuncts to help determine the need for cystoscopy. The combination of 2 tumor markers, NMP22 and 1 cytokeratin (CYFRA 21-1 or urinary bladder cancer antigen), seemed to be the most effective. Further comparative studies are needed to assess the promising diagnostic role of these markers.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; False Positive Reactions; Female; Humans; Keratin-19; Keratins; Male; Middle Aged; Nuclear Proteins; Sensitivity and Specificity; Tissue Polypeptide Antigen; Urinary Bladder Neoplasms

The epithelium in kidneys and urinary bladders contain CK18 as in liver cells. The modulation of cytokeratin 18 during tumor transformation in hepatoma had been previously recognized through a series of biochemical and immunological approaches. A 14 KD hepatoma related molecules was found in the previous studies. We would like to utilize the hepatoma transformation model to study the changes in CK18 in transitional cell carcinoma, using immunoblotting and western blotting techniques. The result is that transitional cell carcinoma retain their CK18 molecule. Furthermore, CK18 related molecules similar to those seen in hepatoma also present in transitional cell carcinoma. The conclusions are transitional cell carcinoma contains CK18 related proteins similar to those seen in hepatoma tissues. We suggest that this element would be responsible for the change during the malignant transformation processes.

Topics: Blotting, Western; Carcinoma, Hepatocellular; Carcinoma, Transitional Cell; Keratins; Kidney Neoplasms; Liver Neoplasms; RNA, Messenger; Urinary Bladder Neoplasms

Topics: Carcinoma, Squamous Cell; Humans; Keratins; Male; Middle Aged; Tomography, X-Ray Computed; Urinary Bladder Neoplasms

Of the 20 known cytokeratins, CK-19 is expressed in normal urothelium, whereas the recently identified CK-20 is expressed in urothelial carcinoma cells but not in normal urothelial cells. The aim of this study was to examine whether CK-20 expression could serve as a noninvasive test in which malignant urothelial cells in urine are detected and monitored.. In the current study, the authors used reverse transcriptase-polymerase chain reaction (RT-PCR) methods to determine the expression of CK-20 in cells separated from the urine of patients with bladder carcinoma. Cells were obtained from the urine of 87 patients divided into the following 2 groups: 1) 14 healthy volunteers without any known history of transitional cell carcinoma (TCC), and 2) 73 patients with hematuria suspected for TCC of the bladder. For control purposes, CK-20 expression was examined in cells of 1) bladder carcinoma tumors of 5 patients, 2) blood of either patients with bladder carcinoma (n = 5) or healthy controls (n = 5), and 3) three different cell lines. RNA of the various cell pellets was extracted and RT-PCR was performed with CK-20 and CK-19 primers (CK-19 was used as a marker for normal epithelial cells).. CK-20 amplification band (370 bp) was obtained with mRNA extracted from TCC cells of either bladder tumor or HT-29 line (a CK-20 colon carcinoma line). Sensitivity of the method was found to be 91%, whereas specificity was 67%. Among the 7 false-positive cases, 3 showed atypia, 3 hyperplasia, and 1 metaplasia, and 2 underwent previously successful TCC tumor removals, suggesting that the CK-20 test also responded to premalignant lesions. No false-positive cases were found in the healthy control group. No other preparation, including blood of the patients of with TCC, showed the CK-20 amplification band.. These results indicate that CK-20 is a potential biomarker for noninvasive detection of bladder carcinoma by assaying uroepithelial cells from the voided urine specimen with RT-PCR.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; False Positive Reactions; Female; Gene Expression Regulation, Neoplastic; Hematuria; HT29 Cells; Humans; Hyperplasia; Keratins; Male; Metaplasia; Middle Aged; Polymerase Chain Reaction; Precancerous Conditions; RNA, Messenger; Sensitivity and Specificity; Transcription, Genetic; Tumor Cells, Cultured; Urinary Bladder; Urinary Bladder Neoplasms; Urothelium

A distinctive variant of a papillary noninvasive transitional cell carcinoma (TCC) of the vagina removed from a postmenopausal woman is described. The neoplasm was evaluated by immunohistochemistry. The designation of this neoplasm as a TCC is supported by its morphological features and its coexpression for cytokeratin (CK) 7 and CK 20. Its main feature is pagetoid infiltration into adjacent vaginal epithelium. This is the second reported case involving a transitional cell metaplasia (TCM) of the vagina, a possible precursor lesion of the TCC.

Topics: Adult; Carcinoma, Transitional Cell; Female; Humans; Immunohistochemistry; Keratins; Metaplasia; Neoplasm Recurrence, Local; Neoplasms, Second Primary; Postmenopause; Urinary Bladder Neoplasms; Vagina; Vaginal Neoplasms

Adenocarcinomas may arise primarily from the urinary bladder, but secondary involvement from adenocarcinomas arising in adjacent organs is more common. In the present study we tried to differentiate primary urinary bladder adenocarcinomas from adenocarcinomas arising from the surrounding organs, based on their antigen profiles in routinely processed, paraffin-embedded tissue specimens. We analysed the staining results using stepwise linear discriminant analysis.. We investigated the usefulness of a panel of antibodies against cytokeratin 7, E48, cytokeratin 20, PSA, PSAP, CEA, vimentin, OC125 and HER-2/neu, to discriminate primary bladder adenocarcinoma from adenocarcinomas arising from the prostate, urachus, colon, cervix, ovary and endometrium. In the differential diagnosis with urinary bladder adenocarcinoma, an overall correct classification was reached for 77% and 81% of urachal and colonic carcinomas, respectively, using CEA, for 93% of prostatic adenocarcinomas using PSA, for 82% and 70% of cervical and ovarian adenocarcinomas, respectively, using OC125, and for 91% of endometrial adenocarcinomas using vimentin. Adding other antibodies did not improve the classification results for any of these differential diagnoses.. For the surgical pathologist, a panel of antibodies consisting of CEA, PSA, OC125 and vimentin is helpful to differentiate primary urinary bladder adenocarcinomas from adenocarcinomas originating from prostate and endometrium, less helpful in differentiation with urachal carcinoma, and not helpful in differentiation with colonic, cervical and ovarian carcinoma.

Topics: Abdominal Neoplasms; Acid Phosphatase; Adenocarcinoma; Antibodies, Monoclonal; Antibody Specificity; CA-125 Antigen; Carcinoembryonic Antigen; Carcinoma, Papillary; Cell Adhesion Molecules; Diagnosis, Differential; Endometrial Neoplasms; Female; Glycoproteins; GPI-Linked Proteins; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratin-7; Keratins; Male; Neoplasms, Unknown Primary; Ovarian Neoplasms; Prostate; Prostate-Specific Antigen; Receptor, ErbB-2; Urachus; Urinary Bladder Neoplasms; Uterine Cervical Neoplasms; Vimentin

Epidermal growth factor (EGF) is excreted in high concentrations in the urine and stimulates urothelial cell growth. The cultured bladder cancer cell line KU-1 was used to study the molecular mechanisms by which EGF affects urothelial tumor growth and invasion activity.. KU-1 cells were grown in cell culture in the presence or absence of EGF. Anchorage-independent cell growth assays and Matrigel invasion assays were performed. Expression of cytokeratins was examined by Northern and Western blot analyses. Chloramphenicol acetyltransferase assays were used to determine whether EGF stimulated matrix metalloproteinase expression.. EGF enhanced anchorage-independent growth in soft agar and increased the number of cells penetrating into a Matrigel membrane. A transient transfection assay revealed that EGF increased the promoter activities of the matrix metalloproteinase 1 and 9 genes in KU-1 cells. Moreover, the morphology of KU-1 cells changed after the addition of EGF to the culture medium. Western and Northern blot analyses demonstrated that EGF decreased cytokeratin 19 expression, but did not affect expression of cytokeratin 8 or 18.. EGF increased the invasive activity of KU-1 bladder cancer cells in part by increasing the secretion of matrix metalloproteinases. Morphologic changes may result from altered composition of cytoskeletal proteins.

Topics: Epidermal Growth Factor; Humans; Keratins; Metalloendopeptidases; Neoplasm Invasiveness; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Humans; Keratin-19; Keratins; Reproducibility of Results; Ultrasonography; Urinary Bladder Neoplasms

Using confocal laser scanning microscopy we studied sections of the T24B, a human bladder carcinoma, grown in C.B.-17 scid/scid or NMRI nu/nu mice in order to examine the relationship between tumor tissue and tumor vessels. Tumor cells were labelled with FITC-anti-cytokeratin and blood vessel endothelia with Cy3-labelled BS-I lectin. In contrast to our expectation, no major leaks in the endothelial lining of blood vessels were observed. We are looking for a suitable marker for mouse lymphatics in order to investigate their possible role.

Topics: Animals; Endothelium, Vascular; Fluorescent Antibody Technique, Indirect; Humans; Keratins; Mice; Mice, Nude; Mice, SCID; Microscopy, Confocal; Microscopy, Electron, Scanning; Neoplasm Transplantation; Transplantation, Heterologous; Tumor Cells, Cultured; Urinary Bladder Neoplasms

The present study was designed to evaluate the role of the cytokeratin-19 fragment (CYFRA 21-1) as a tumor marker in bladder cancer. The bladder cancer cell line JMSU1 was used in the preclinical study. Subjects comprised 120 healthy volunteers, 20 patients with benign bladder diseases including acute cystitis and bladder stones, 12 patients with chronic renal failure, and 117 patients with histologically confirmed primary bladder cancer. CYFRA 21-1 concentrations were measured by a sandwich enzyme-linked immunosorbent assay. The preclinical study in vitro and in vivo showed that JMSU1 produced and released CYFRA 21-1 in the culture supernants and serum of JMSU1-bearing nude mice. The cutoff level was set at 3.5 ng/ml (mean + 3 SD) by the analysis in the healthy volunteers. Under this condition, sensitivity was 0% in benign bladder diseases, 83% in chronic renal failure, and 41.9% in bladder cancer. Serum CYFRA 21-1 levels increased significantly as tumor stage advanced or tumor grade increased. A serial follow-up study demonstrated that patients with progressive showed a gradual increase in serum CYFRA 21-1 levels while patients who responded to the treatments had a marked decrease in serum CYFRA 21-1 levels. Higher serum CYFRA 21-1 levels were related to poor survival. The present study suggest that serum CYFRA 21-1 is a useful marker to monitor the clinical course of bladder cancer and to provide prognostic information.

Topics: Adult; Aged; Aged, 80 and over; Animals; Antigens, Neoplasm; Biomarkers, Tumor; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratin-19; Keratins; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Middle Aged; Neoplasm Transplantation; Sensitivity and Specificity; Tumor Cells, Cultured; Urinary Bladder Diseases; Urinary Bladder Neoplasms

We report 25 sarcomatoid carcinomas of the urinary bladder with a prominent myxoid and/or sclerotic appearance. The average age of the patients was 72 years (range, 50-92 yr); 14 were men, and 11 were women. The cystoscopic appearance varied from a large polypoid mass to an intramural mass with bladder wall thickening, often with necrosis and ulceration. The tumors ranged from 3 to 10 cm and were typically rubbery or gelatinous with a brown, pink, or gray color. Microscopy revealed tapering spindle cells with a variable admixture of cohesive non-spindled cells. Twenty-two cases had an invasive overtly epithelial carcinomatous component, and in situ transitional carcinoma was present in 12 cases. All of the cases had areas with myxoid change, ranging from extensive to focal, separating the spindle cells. Fourteen cases had areas of sclerosis. In all the cases, the spindle cells were atypical, at least focally, with hyperchromatic pleomorphic nuclei, prominent nucleoli, and coarse chromatin. Mitotic activity was prominent in the majority of cases, and abnormal mitotic figures were frequent. In eight cases, the myxoid histologic pattern was very reminiscent of an inflammatory pseudotumor, a diagnosis frequently entertained and erroneously made in one case; many of the spindle cells in three of these cases were mildly atypical, with minimal mitotic activity. The spindle cells were immunoreactive for cytokeratin (12 of 19), vimentin (16 of 17), carcinoembryonic antigen (3 of 15) and muscle-specific actin (4 of 16), and nonreactive for epithelial membrane antigen, desmin, S-100, KP1, CD34, and Leu-M1. The epithelioid carcinomatous areas were highlighted by the cytokeratin immunostain. These features and the conventional light microscopic features indicative of a diagnosis of carcinoma distinguish this tumor from reactive of neoplastic mesenchymal lesions.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Diagnosis, Differential; Female; Granuloma, Plasma Cell; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Urinary Bladder Neoplasms; Vimentin

A case of a 66-year-old Japanese woman with two synchronous urinary bladder tumors, namely verrucous carcinoma and poorly differentiated squamous cell carcinoma, is described. Both tumors were accompanied by widespread squamous metaplasia in the background and unassociated with bilharzial infection. The poorly differentiated squamous cell carcinoma, having a satellite tumor in its proximity, was large and in an advanced stage. The verrucous carcinoma was small with minimal invasion to the muscularis propria. The boundary between both tumors was well defined, suggesting colliding growth appearance. Immunoexpression of cytokeratins of verrucous carcinoma was similar to that of squamous metaplasia, and significant differences between verrucous carcinoma and poorly differentiated squamous cell carcinoma were demonstrated in their immunoexpression of cytokeratins.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Neoplasms, Multiple Primary; Schistosomiasis; Urinary Bladder Neoplasms

Vulvar Paget's disease (VPD) is the most common type of extramammary Paget's disease; however, the frequency of dermal invasion and its clinical significance are unclear, as are the frequency and relationship of an associated regional internal cancer. Thus, we studied the clinicopathologic and immunohistochemical features of 19 patients with VPD. Patients ranged in age from 56 to 86 years (median 65). VPD was entirely intraepithelial (IE-VPD) in 13 patients. Three patients developed IE-VPD recurrence and one developed deeply invasive and metastatic VPD at 10.8 years. Five patients had invasive Paget's disease (INV-VPD) characterized by clinically occult microscopic foci of superficial dermal invasion, ranging in depth from 0.3 to 0.9 mm. All five patients were alive without disease after 12 months to 17 years (median 66 months). A regional internal cancer (CA ASSOC-VPD) occurred in one patient whose VPD was preceded by a deeply invasive grade 3 transitional cell carcinoma of the urinary bladder 9 months earlier. Immunophenotypes of 16 cases with IE-VPD or INV-VPD were CK7+/CK20-/GCDFP15+ in 14 cases and CK7+/CK20+/GCDFP15+ in two cases, with concordant immunophenotypes of the intraepithelial and invasive components in all cases studied. The patient with CA ASSOC-VPD had a CK7+/CK20+/GCDFP15- immunophenotype in the invasive TCC of the urinary bladder and the VPD. We conclude that the predominant form of VPD begins as a primary cutaneous intraepithelial neoplasm that is universally CK7+/GCDFP15+. Foci of unsuspected synchronous dermal invasion by Paget's cells can be expected in almost one third of cases. Subsequent progression into an invasive carcinoma occurs less often. Foci of "minimally invasive" carcinoma (<1 mm) probably do not adversely affect prognosis, whereas deeply invasive carcinoma behaves as a fully malignant adenocarcinoma. The rarer form of VPD appears to result from secondary intraepithelial spread from an associated regional internal carcinoma. The finding of Paget's cells that are CK20+/GCDFP15- suggests the presence of a regional internal carcinoma with a corresponding immunophenotype.

Topics: Aged; Aged, 80 and over; Apolipoproteins; Apolipoproteins D; Biomarkers, Tumor; Carcinoma, Transitional Cell; Carrier Proteins; Female; Glycoproteins; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratin-20; Keratins; Membrane Transport Proteins; Middle Aged; Neoplasm Invasiveness; Paget Disease, Extramammary; Urinary Bladder Neoplasms; Vulvar Neoplasms

An 82-year-old man with microhematuria and class V cells in his urinary cytologic specimen was referred to our clinic. Cystoscopy revealed a solid, broad-based tumor of 1 cm in diameter in the bladder diverticulum. A partial cystectomy was performed. The tumor was composed of transitional cell carcinoma (TCC), sarcomatous spindle cells and the area of transition between TCC and spindle cells. The histopathological diagnosis was sarcomatoid carcinoma. Immunohistochemical examination showed that the spindle cells and the area of transition were positive for keratin, cytokeratin, vimentin and muscle actin. The histopathological and immunohistochemical transitions between the TCC and the spindle cells suggested that the sarcomatous elements originated from the TCC during tumor progression.

Topics: Actins; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Transitional Cell; Humans; Immunohistochemistry; Keratins; Male; Sarcoma; Urinary Bladder Neoplasms; Vimentin

Presented is a case report of a urinary bladder carcinoma that had an unusual morphology and phenotype. A 65-year-old Japanese man complained of gross hematuria. Cytological examination of the urine before a partial cystectomy revealed small, round atypical cells with a high nuclear/cytoplasmic ratio, scant cytoplasm, and hyperchromatic nuclei with coarse and granular chromatin in a bloody background. Several tumor cells had relatively large and vesicular nuclei with prominent eosinophilic nucleoli and obscure perinucleolar halos. A small number of large atypical urothelial cells were also recognized. The tumor recurred locally 3 months after the operation. The urine cytology during recurrence showed the same features without the atypical urothelial cells. These cytological findings suggested a case of small cell undifferentiated carcinoma (SCUC) combined with transitional cell carcinoma (TCC). An histology of the resected specimen before the recurrence revealed that the SCUC was consistent with a variant type of SCUC proposed for the lung and showed transition with TCC in situ. M-VAC chemotherapy after a total cystectomy was less effective. The patient died 6 months after diagnosis. A variant subtype of SCUC of the urinary bladder associated with TCC in situ has not been previously reported. Although this histological type is very rare, its earlier cytological detection is needed for appropriate therapy.

Topics: Aged; Biomarkers, Tumor; Carcinoma; Carcinoma in Situ; Carcinoma, Transitional Cell; Cytodiagnosis; Fatal Outcome; Humans; Immunohistochemistry; Keratins; Lewis X Antigen; Male; Microscopy, Electron; Neoplasm Recurrence, Local; Urinary Bladder Neoplasms

The presence of squamous differentiation in transitional cell carcinomas has been variably related to prognosis and response to radiotherapy. This study sought to establish whether cytokeratin (CK) 14, normally expressed in the basal cells of squamous epithelium, could be used as a reliable marker of the emergence of a squamous phenotype in transitional cell carcinomas. In a series of 42 tumours, CK14 was expressed in areas of squamous morphology, whereas CK20 identified continuing urothelial differentiation. Furthermore, focal positivity for CK14 was present in a proportion of tumours with no morphological evidence of squamous differentiation, suggesting that it is a more sensitive marker of phenotypic switch. Investigation of CK subtypes may be more powerful than morphology alone in clinical studies of transitional cell carcinomas as CK expression profiles have been related to treatment response in other tumour types.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cell Differentiation; Humans; Immunohistochemistry; Keratins; Kidney Neoplasms; Prognosis; Ureteral Neoplasms; Urinary Bladder Neoplasms; Urologic Neoplasms

Topics: Antigens, Nuclear; Biomarkers; Carcinoma, Transitional Cell; DNA, Neoplasm; Flow Cytometry; Humans; Immunohistochemistry; Keratin-7; Keratins; Ki-67 Antigen; Nuclear Proteins; Ploidies; S Phase; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

Schistosomiasis remains one of the major public health problems of the tropics. Conservative estimates place the number of infected individuals at about 200 millions. In Egypt, carcinoma of the urinary bladder associated with schistosomiasis is the foremost oncologic problem, because of its high frequency and the late presentation of cases. A newly developed monoclonal antibody CK1K10 to keratinized grade 1 squamous cell carcinoma was used in a dot enzyme-linked immunosorbent assay (Dot ELISA) to test urine samples of 118 patients with bladder carcinoma, 291 patients with genitourinary pathology other than bladder carcinoma, in addition to 550 healthy controls. The overall sensitivity of the dot ELISA was 90% among 118 patients with bladder carcinoma. Twenty-seven of 33 transitional cell carcinoma cases (82%), 68 of the 71 squamous cell carcinoma cases (96%), 7 of 10 undifferentiated tumors cases (70%), and 4 of 4 adenocarcinoma were positive with this assay. The specificity was 90% in our sample population. A comparative study of diagnosis by cytology and dot ELISA was carried out in 57 patients with bladder carcinoma. Dot ELISA was found to be superior as a screening tool for high risk groups (P < .001 using chi-square test). Cytology detected 21% of transitional cell carcinoma, 68% of squamous cell carcinoma, 50% of adenocarcinoma, and 86% of undifferentiated tumors. The dot ELISA assay should be useful for screening high-risk groups because it does not require sophisticated equipment, is noninvasive, does not require highly trained staff, and can be performed in less than 30 minutes.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cytodiagnosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Pregnancy; Risk Factors; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urine

Walthard cell nests, the Brenner tumor (benign, proliferating, low malignant potential, and malignant), and primary ovarian transitional cell carcinoma are considered to be primary female genital tract proliferations of transitional-type (urothelial) epithelium on conventional light microscopic grounds. In order to further investigate the similarities (or dissimilarities) of proliferations of female genital tract transitional epithelium and urothelium, we compared transitional cell proliferations (TCPs) of the female genital tract (n = 25) and urinary bladder (n = 15) using antibodies to carcinoembryonic antigen (CEA; clone 0062), carbohydrate determinant 19-9 (CA19-9; clone 1116-NS-19-9), cytokeratin 7 (CK-7; clone OV-TL 12/30), and cytokeratin 20 (CK-20; clone Ks 20.8), four monoclonal antibodies that have been shown to stain transitional cell urothelial proliferations. Both groups of tumors exhibited significant staining for CEA, CA19-9, and CK-7, and the difference in numbers of cases staining was not significant. CA19-9 was present in 15 of 25 female genital tract TCPs as compared with 12 of 15 bladder TCPs; CEA was present in 17 of 25 female genital tract TCPs and nine of 15 comparable bladder TCPs. CK-7 was present in all cases studied with the exception of one Walthard cell nest and a malignant Brenner tumor that was not immunoreactive with the other antibodies tested. In contrast, 13 of 15 bladder TCPs were CK-20 positive, whereas only one of 25 female genital tract TCPs was positive (< 5% of cells). Walthard cell nests and benign Brenner tumors were more likely to be CA19-9 positive than were Brenner tumors of low malignant potential, malignant Brenner tumors, and primary transitional cell carcinoma of the ovary. We conclude that despite their apparent morphologic and immunologic similarity to TCPs of the urinary bladder (particularly at the histologically low-grade end of the transitional cells spectrum), Walthard cell nests and ovarian Brenner tumors constitute an immunophenotypically distinct form of TCP.

Topics: Antibodies, Monoclonal; Brenner Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Carcinoma, Transitional Cell; Female; Humans; Immunohistochemistry; Keratins; Ovarian Neoplasms; Precancerous Conditions; Urinary Bladder Neoplasms

The determination of tumor markers in urine samples has been proposed as an effective diagnostic tool in bladder cancer. The aim of the present investigation was to validate in urine samples the assay of the CYFRA21.1 cytokeratin-related marker, the serum concentrations of which showed promising diagnostic utility in patients with bladder cancer. First-voided urine samples were collected from patients with different malignancies. CYFRA21.1 was assayed with a commercially available enzyme immunoassay (Boehringer Mannheim). Different centrifugation patterns, the use of different buffers and nonionic detergents, and pH variations were evaluated. We demonstrated that: (a) cells and cell debris contain a large amount of CYFRA21.1 and must be eliminated by centrifugation; (b) storage at -20 degrees C causes amorphous precipitate, which may aspecifically bind CYFRA21.1; (c) the latter behavior may be prevented by diluting fresh urine samples with phosphate buffer with nonionic detergent added; (d) pH variations within the range 4.9-8.2 do not significantly affect CYFRA21.1 assay results. Provided that samples are diluted with buffer containing nonionic detergent, the CYFRA21.1 assay showed good precision and accuracy characteristic in urine samples. We therefore propose a standard protocol for the collection of urine samples for CYFRA21.1 assay. In a preliminary clinical evaluation, CYFRA21.1 concentrations in 16 patients with primary bladder cancer were higher than in healthy subjects. In the urine collected in the follow-up of patients treated for bladder cancer, CYFRA21.1 tended to be higher in relapsed patients than in those without evidence of disease. These preliminary data induced us to extend the clinical trial to establish the actual role of this assay in routine use.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Buffers; Centrifugation; Detergents; Freezing; Humans; Hydrogen-Ion Concentration; Immunoenzyme Techniques; Keratin-19; Keratins; Neoplasm Recurrence, Local; Reproducibility of Results; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urologic Diseases

Circulating cancer cells in the blood play a central role in the metastatic process. Their number can be very small and techniques for their detection need to be both sensitive and specific. Polymerase chain reaction (PCR) has been successfully used to detect small numbers of tumour cells in haematological cancer in which abnormalities in DNA are sufficiently consistent to make this possible. For most solid tumours this not yet feasible. However, we have found that reverse transcriptase (RT)-PRC for tissue-specific gene expression is a useful technique for identifying small numbers of circulating cells in melanoma and neuroblastoma patients. In this report we describe detection of colon carcinoma cells by RT-PCR using CK 20 mRNA as a marker. Unlike other cytokeratin genes examined (CK 8 and CK 19), CK 20 was not transcribed in normal haematopoietic cells. This suggests a role for RT-PCR in the detection of colon carcinoma metastasis in blood and bone marrow, using CK 20 as the target gene. Future analysis of clinical material will determine the clinical significance of this technique.

Topics: Adenocarcinoma; Base Sequence; Biomarkers, Tumor; Bone Marrow; Breast Neoplasms; Carcinoma, Transitional Cell; Colonic Neoplasms; Epithelium; Humans; Keratins; Molecular Sequence Data; Neoplasm Metastasis; Neoplasm Proteins; Neoplastic Cells, Circulating; Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Sensitivity and Specificity; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Epithelial differentiation antigens have been correlated with morphologic differentiation of neoplastic urothelium. Moreover, epidermal growth factor, which is a polypeptide regulating growth and differentiation of normal and neoplastic cells, is found in high concentrations in the urine while its receptors (EGFR) have been identified in bladder tumors. The aim of this study was to investigate the immunohistochemical expression of cytokeratin, epithelial membrane antigen (EMA), CEA and EGFR in transitional cell bladder carcinomas (TCC) and to define any correlation of their expression with tumor grade, stage and patient survival. Twenty-four biopsy specimens obtained from patients with TCC were studied retrospectively. There were 23 men and 1 woman with a mean follow-up of 64 months. Eight biopsy specimens, which represented tumor recurrences of 4 patients, were also included in our material. The immunohistochemical avidin-biotin complex method was performed on paraffin sections for the detection of cytokeratin and EGFR with monoclonal antibodies as well as CEA with a polyclonal antibody. Cytokeratin was detected in 83.5% of the TCC, EMA in 62% and CEA in 70%. The expression of the epithelial differentiation antigens in TCCs was heterogenous, showing an increased incidence in high-grade and high-stage TCC. The CEA expression in TCC demonstrated a statistically significant correlation with patient survival (p < 0.02). EGFR was detected in 50% of the TCC. Although not statistically significant, a trend was found for a higher percentage of EGFR detection in high-grade TCC. EGFR expression was significantly associated with tumor stage and patient survival (p < 0.01 and p < 0.04, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Aged, 80 and over; Antigens, Differentiation; Carcinoembryonic Antigen; Carcinoma, Transitional Cell; Epithelium; ErbB Receptors; Female; Humans; Keratins; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; Mucins; Neoplasm Staging; Prognosis; Urinary Bladder Neoplasms

We have induced tumors by feeding guinea pigs with a diet containing 25 or 30% dried bracken fern for 100 or 150 days. A high incidence of bladder tumors was obtained. All but one animal had preneoplastic or neoplastic lesions after 4 months; after one year, 24 or 25 exposed animals had carcinoma. Bladder tumors obtained were essentially pure transitional cell carcinomas, although 4 cases (7% of the exposed animals and 10% of the 39 transitional cell carcinoma observed) showed areas of focal squamous metaplasia. Immunohistological detection of cytokeratins 10, 13, and 18 confirmed the transitional nature of these tumors. Tumor development can be followed by ultrasonography and cytology. Bladder tumors arose through several steps. Dysplasia and preneoplastic hyperplasia were seen after 4 months and papillary carcinomas appeared after 6 months, whereas muscle-invasive carcinomas required 1 year. Thus this model reproduces the full spectrum of preneoplastic and neoplastic bladder lesions observed in humans. Interestingly, when tumors were induced in older guinea pigs, none of them progressed to a muscle-invasive stage. This phenomenon should provide the opportunity to study the molecular mechanisms associated with these two different growth patterns, a major issue in understanding human bladder tumor progression.

Topics: Animals; Carcinoma, Transitional Cell; Diet; Disease Models, Animal; Epithelium; Female; Follow-Up Studies; Guinea Pigs; Humans; Immunohistochemistry; Keratins; Male; Neoplasm Invasiveness; Plants; Urinary Bladder; Urinary Bladder Neoplasms

To determine p53 expression in cells in bladder washings and to relate this to DNA content and clinical outcome.. Washings from 102 patients (41 with newly diagnosed superficial tumours [pTa and pT1], 49 with recurrent superficial tumours and 12 with carcinoma invading bladder muscle) were studied. In 39 cases, the primary bladder tumour was also analysed. The rates of tumour recurrence and progression were determined for the new superficial tumours and related to both p53 expression and DNA content.. Cells positive for p53 were detected in 22 of 90 (24%) washings from patients with superficial bladder cancer. P53 expression correlated with tumour stage (P < 0.05), grade (P < 0.05) and abnormal DNA content (P < 0.05). The analysis of pure urothelial (cyto-keratin-positive) cells improved the detection of DNA abnormalities (P < 0.001). In 74% of cases where both washings and tumour were analysed, the results for DNA content agreed. Of 41 new superficial tumours, 27 (66%) recurred (11 were p53-positive, 16 were p53-negative, P = 0.221; 17 had abnormal DNA content, 10 were diploid, P = 0.069). Four patients progressed (one was p53-positive, P = 0.315 and all had abnormal DNA content, P = 0.072).. P53-positive cells can be detected in washings using flow cytometry and were more commonly detected in association with aneuploid tumours. At short-term follow-up, flow cytometric analysis of DNA content in washings had greater predictive value than had p53 expression. Few washings contained aneuploid cells when the primary tumour contained diploid cells, although the collection of washings is a convenient way of sampling tumour cells.

Topics: Aged; Carcinoma in Situ; Disease Progression; DNA, Neoplasm; Flow Cytometry; Follow-Up Studies; Humans; Keratins; Neoplasm Recurrence, Local; Ploidies; Tumor Suppressor Protein p53; Urinary Bladder Neoplasms

The display of repertoires of antibody fragments on the surface of filamentous phage offers a new way to produce immunoreagents with defined specificities. Here we report the selection of antibody fragments against the cytoskeletal fraction of T24 bladder cancer cells. To focus selection to a specific antigen, we eluted bound phage with a mouse monoclonal antibody directed against cytokeratins. Our initial studies proved that such a selection procedure with a library, carrying the mouse antibody fragment repertoire, resulted in phage specificity for the antigen against cytokeratin 8, recognized by the mouse monoclonal antibody. To facilitate detection of reactive clones, monoclonal antibodies against phage epitopes were developed. A human synthetic library (> 10(8) clones) was used for selection by competitive elution after binding to T24 cytoskeleton. About 50% of the phage reacted in ELISA with cytoskeletons of T24 cells, while with noncytokeratin containing cells no reaction was observed. Immunofluorescence studies and Western blotting with a number of these clones showed reactivity against cytokeratin. We conclude that the competitive elution method can be used as a rapid technique to obtain immunoreactive phages, and eventually human single chain antibodies directed against defined epitopes which were formerly characterized and validated by mouse monoclonal antibodies.

Topics: Animals; Antibodies, Monoclonal; Bacteriophages; Enzyme-Linked Immunosorbent Assay; Humans; Keratins; Mice; Mice, Inbred BALB C; Tumor Cells, Cultured; Urinary Bladder Neoplasms

To study the clinical, histological, and immunohistochemical findings of small cell carcinoma (SCC) of the urinary bladder, and also to delineate its behaviour in comparison with transitional cell carcinomas of the bladder.. A retrospective review of 552 patients with bladder cancer yielded six cases (1%) of small cell carcinoma which were histologically identical to pulmonary small cell anaplastic carcinoma. Clinical data and follow-up were collected. Aside from the conventional histological parameters, an immunohistochemical study with AE1-AE3 and Cam 5.2 keratins, epithelial membrane antigen, neuron-specific enolase, chromogranin, synaptophysin, ACTH, calcitonin, and prostatic specific antigen was performed.. The clinical presentation did not differ from conventional transitional cell carcinoma, haematuria being the most frequent complaint (four cases). All the cases presented as flat tumours. On light microscopy, there were oat cell (four cases), intermediate (one case) and mixed oat-cell/intermediate (one case) variants. Three cases were associated with transitional cell carcinoma. Dysplastic changes were observed in the adjacent urothelium in one case only. At the time of diagnosis, all tumours were deeply invasive (pT3). Three cases were Stage III and three Stage IV, with involvement of regional lymph nodes and metastases to the liver (two cases) and lung (one case). Immunohistochemically, epithelial markers were variably expressed as AE1-AE3 keratin (5/6), Cam 5.2 keratin (2/6) and epithelial membrane antigen (3/6). Neuron specific enolase was demonstrated in every case. Chromogranin, however, was expressed in only one case. Synaptophysin, ACTH, calcitonin, and prostatic specific antigen all gave negative results. All the patients died of the disease and the overall length of survival was very poor (range 5-25 months, mean 13.3).. Small cell carcinomas show the same histological patterns as their pulmonary counterpart. Immunohistochemistry reveals a wide spectrum of activity, enolase and keratins being the most constant. The present study confirms that the overall prognosis of this tumour is very poor.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Small Cell; Carcinoma, Transitional Cell; Humans; Immunohistochemistry; Keratins; Male; Phosphopyruvate Hydratase; Retrospective Studies; Urinary Bladder Neoplasms

Sarcomatoid carcinoma is a rare tumor in the urinary bladder and accounts for approximately 0.3% of all bladder malignancies. In this study, the clinicopathologic findings of 18 cases are described. Distribution of sex and age and clinical symptoms are not distinctive from transitional cell carcinoma. The tumor behaves as a high-grade malignancy with advanced initial stage and unfavorable outcome. Surgery is the therapy of choice. Histological differentiation from true sarcoma may be difficult. Recognition rests on the co-existence of an overt carcinomatous component or demonstration of the epithelial nature by immunohistochemistry or electron microscopy.

Topics: Aged; Aged, 80 and over; Carcinoma; Female; Follow-Up Studies; Humans; Immunohistochemistry; Keratins; Male; Membrane Glycoproteins; Microscopy, Electron; Middle Aged; Mucin-1; Urinary Bladder Neoplasms; Vimentin

After extraction in high salt buffers and Triton X-100, cytokeratin polypeptides could be isolated from normal epithelium and transitional cell carcinoma of the human bladder in the southwestern coast of Taiwan. The polypeptides were solubilized via lysis buffer, separated by two-dimensional gel electrophoresis, and identified by immunoblot. Five cases of normal epithelium expressed cytokeratin No. 5 (58 kd), 7 (54 kd), 18 (45 kd) and one case further expressed cytokeratin No. 19 (40 kd) in addition to the above cytokeratins. Combined patterns of cytokeratin polypeptides were observed among various Grades of transitional cell carcinoma of bladder. Comparing the patterns between the normal epithelium and the transitional cell carcinoma of bladder, 88% (14/16) of carcinomas expressed cytokeratin No. 8 (52.5 kd) and 63% (10/16) expressed No. 17 (46 kd), which were not seen in the normal epithelium. Some other cytokeratin polypeptides particular to carcinomas were No. 4 (59 kd), No. 10 (56.5 kd), No. 13 (54 kd) and No. 14 (50 kd) and were only present in less than 50% (ranging from 31-56%) of the examined specimens. Thus cytokeratin No. 8 and 17 could be helpful markers for diagnosis of bladder carcinoma.

Topics: Carcinoma, Transitional Cell; Epithelium; Humans; Keratins; Urinary Bladder; Urinary Bladder Neoplasms

Double immunoenzymatic labelling made possible the simultaneous staining of two antigens with a mixture of polyclonal and monoclonal commercial antibodies. Immunocharacterization of intermediate filament proteins was found to be an accurate indicator of histogenesis in urinary bladder tumours of cattle.

Topics: Animals; Antibodies, Monoclonal; Cattle; Cattle Diseases; Desmin; Immunoenzyme Techniques; Intermediate Filaments; Keratins; Urinary Bladder Neoplasms; Vimentin

A phage-display combinatorial library of VL and VH sequences of mouse antibodies was constructed, which contained 4.5 x 10(7) independent clones. From this library pools of phage were selected by up to four biopanning rounds on cytoskeletal preparations of ovarian carcinoma cells (OVCAR-3). Phage of these pools were then allowed to bind to a cytoskeleton preparation of bladder carcinoma cells (T24). The binding phage were challenged by a monoclonal antibody (mAb) directed against an epitope on cytokeratin 8. Displaced phage were rescued and screened for anti-cytokeratin immunoreactivity by ELISA, indirect immunofluorescence and Western blotting. About 50% of the phage selected by competition with the cytokeratin mAb reacted with the cytoskeletal preparations of T24 cells in ELISA. In contrast, in non-cytokeratin-containing cells, no reaction was observed. Immunofluorescence and Western blotting studies with a number of these clones showed reactivity against cytokeratin. We conclude that the phage-display competitive elution method can be used as a rapid technique to obtain immunoreactive phages, and eventually single-chain Fv (scFv) antibodies directed against defined epitopes, which were formerly characterized and validated by mAbs.

Topics: Antibodies, Monoclonal; Bacteriophages; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Fluorescent Antibody Technique; Humans; Immunoassay; Keratins; Recombinant Proteins; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Topics: Antibodies; Biomarkers, Tumor; Breast Neoplasms; Cell Separation; DNA, Neoplasm; Female; Flow Cytometry; Humans; Intermediate Filament Proteins; Keratins; Molecular Probes; Neoplasms; Ploidies; Staining and Labeling; Urinary Bladder Neoplasms

Topics: Humans; Keratins; Leukoplakia; Male; Metaplasia; Middle Aged; Urinary Bladder Neoplasms

Pseudosarcomatous fibromyxoid tumor of the genitourinary tract is a rare pathologic entity of hitherto unknown etiology that, because of the cellular pleomorphism and the infiltrative nature of the lesion, may be mistakenly diagnosed as sarcomatoid carcinoma or sarcoma. We retrospectively studied nine pseudosarcomatous fibromyxoid tumors involving the bladder and prostate to define characteristic parameters that may allow for accurate diagnosis. The study patients included four men and five women with a mean age of 48.7 years. Histologic analysis revealed myxoid lesions with a proliferation of spindle fibroblastic cells in a background of granulation tissue-type vascularity and inflammatory cells. Mitoses were infrequent and no atypical forms were found. Immunostaining was positive for vimentin and smooth muscle actin, and negative for S-100 protein, desmin, myoglobin, and keratin. Ultrastructurally, the lesion displayed fibroblastic and myofibroblastic cell features. Flow cytometric DNA content analysis revealed uniform DNA diploidy and a low S-phase fraction. All patients were alive and well with no evidence of disease after a mean follow-up of 4.8 years. In contrast, the sarcomatoid carcinomas and sarcomas of the urinary bladder and prostate that were used as controls occurred in older patients and had more frequent mitoses with atypical forms, tumor-type necrosis, and different immunostaining profiles; they were preponderantly aneuploid or diploid with high S-phase fraction. Awareness of the clinicopathologic and biologic characteristics of these lesions is necessary to ensure their accurate diagnosis and to prevent unnecessary radical therapy.

Topics: Actins; Adult; Carcinoma; Desmin; Diagnosis, Differential; DNA, Neoplasm; Female; Fibroma; Flow Cytometry; Humans; Immunohistochemistry; Keratins; Male; Microscopy, Electron; Middle Aged; Myoglobin; Ploidies; Prostatic Neoplasms; Retrospective Studies; S Phase; S100 Proteins; Sarcoma; Urinary Bladder Neoplasms; Vimentin

In this study, we attempted to find a gene or genes which were differentially expressed between a non-tumorigenic rat bladder cell line and a highly tumorigenic/metastatic bladder carcinoma cell line that was derived from the former after treatment in vitro with N-methyl-N-nitrosourea. We cloned a rat keratin 5 cDNA by a differential hybridization technique and found that all of the non-tumorigenic cells (7/7) and normal bladder tissue expressed keratin 5, but most of the tumorigenic cells (8/10) did not express keratin 5. Furthermore, in a spontaneously transformed cell line, keratin 5 expression was lost during the transformation process. These results suggest that loss of keratin 5 expression is closely associated with acquisition of a tumorigenic phenotype by rat bladder non-tumorigenic cells.

Topics: Animals; Base Sequence; Biomarkers, Tumor; Cell Transformation, Neoplastic; Cloning, Molecular; DNA Primers; Epithelial Cells; Epithelium; Gene Expression Regulation, Neoplastic; Gene Library; Keratins; Male; Mice; Mice, Nude; Molecular Probe Techniques; Molecular Sequence Data; Rats; Tumor Cells, Cultured; Urinary Bladder Neoplasms

The possibility of using cytokeratin antibodies for the radioimmunolocalization of urinary bladder cancer was studied. A monoclonal murine IgG antibody was raised against cytokeratin 8 and labelled with iodine-125; normal murine IgG was used for control purposes. The urothelial cancer cell line RT4 was transplanted into immunodeficient nude mice. The anti-cytokeratin 8 antibody was administered intraperitoneally and its uptake in the tumour and other organs was analyzed with a computerized gamma camera. Optimal scintigraphic visualization occurred 11 days after antibody administration. The tumour/blood ratio of the specific antibody was 5.64 (+/- 5.01 SD) on day 11, compared with 0.73 (+/- 0.35 SD) in the control. Autoradiography demonstrated antibody uptake preferentially in viable sections of the tumour. The antibody uptake is presumed to be the result mainly of binding to the released cytokeratin in and around cells lysed during natural cellular death. The monoclonal murine anti-cytokeratin antibody is of potential interest in studies aimed at improving the clinical staging of urinary bladder cancer.

Topics: Animals; Antibodies, Monoclonal; Autoradiography; Humans; Immunoenzyme Techniques; Iodine Radioisotopes; Keratins; Mice; Mice, Nude; Neoplasm Transplantation; Radioimmunodetection; Tomography, X-Ray Computed; Urinary Bladder Neoplasms

Inverted papilloma (transitional cell papilloma, inverted type) is a rare, benign urothelial tumor. The distribution of cytokeratin (CK) expression in 22 cases was investigated and compared with normal urothelium and urothelial carcinomas: CK7/8, basal increased positivity; CK13, diffuse positivity; CK18, loss of intensity and loss of umbrella cell staining; CK19, reduction of positivity; CK20, reduction of umbrella cell staining. The data indicate that the inverted papilloma is a basal cell urothelioma.

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Cystectomy; Electrosurgery; Humans; Immunoenzyme Techniques; Keratins; Neoplasms, Second Primary; Papilloma; Papilloma, Inverted; Urinary Bladder Neoplasms

The expression of cytokeratins (CK) 1, 4, 5/6, 8, 13, 18, 19 and 20 and involucrin in 42 cases of squamous cell carcinomas from various locations was examined. The tumours expressed CK5/6 in 55%, CK8 in 76%, CK13 in 43% and CK19 in 95% of cases. The CK5/6-positive primary tumours were from uterine cervix, head and neck, lung, skin, oesophagus and urinary bladder, and the CK13-positive primary tumours were from uterine cervix, lung and vulva. Metastatic squamous cell carcinomas from head and neck more frequently expressed CK5/6 and 13, 7/7 (100%) and 6/7 (86%) compared with 3/5 (60%) and 0/5 (0%) in the primary squamous cell carcinomas. Few cases were CK1, CK4 and CK18 immunoreactive. CK20 immunoreactivity was not observed. Involucrin was expressed in 71% of tumours, and most of the involucrin-positive cells were located at the central parts of tumour cell clusters except for one case in which the peripheral cells around tumour cell clusters were positive. Thus, expression of the so-called simple epithelial markers CK8 and CK19 occurs in the majority of squamous cell carcinomas. The absence of CK20 immunoreactivity may be helpful in differential diagnosis.

Topics: Carcinoma, Squamous Cell; Female; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Lymphatic Metastasis; Mouth Neoplasms; Protein Precursors; Skin Neoplasms; Urinary Bladder Neoplasms; Uterine Neoplasms

Bladder washing specimens containing inflammatory or squamous cells have been difficult to accurately analyze with single-parameter DNA flow cytometric (FCM) methods.. The anticytokeratin 18 antibody, CK5, was used in a multiparameter assay of 275 bladder washing and voided urine specimens to immunoselect only the bladder transitional cells for DNA analysis.. Flow cytometric detection of transitional cell carcinoma was increased by immunoselection of CK5-positive cells in specimens from patients with disease. Unfortunately, a similar increase in hyperdiploid cells in pathologically benign specimens was observed, which resulted in a false-positive rate of 45%. In some instances, multiparameter FCM assays with CK5 could detect aneuploid cell populations not clearly evident by single-parameter analysis.. However, the results from this study of the hyperdiploid cell fraction showed that the increased sensitivity resulting from the use of CK5 was not clinically useful because of the decrease in specificity.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Neoplasm; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; DNA, Neoplasm; Flow Cytometry; Humans; Keratins; Male; Mice; Urinary Bladder; Urinary Bladder Neoplasms

The clinicopathological features of two carcinosarcomas of the urinary bladder are reported. The tumours occurred in a 64- and a 66-year-old patient presenting with haematuria and both were polypoid. The epithelial component was consistent with small cell undifferentiated carcinoma with neuroendocrine differentiation, whereas the sarcomatous component did not display specific features. The carcinomatous component showed immunohistochemical reactivity for different epithelial markers as well as for chromogranin and neuron specific enolase. Conversely, the sarcomatous cells stained strongly for vimentin and in one case for muscle actin and smooth muscle actin. The differential diagnosis of biphasic tumours of the bladder is discussed and the literature reviewed.

Topics: Actins; Aged; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinosarcoma; Chromogranins; Diagnosis, Differential; Female; Humans; Keratins; Male; Middle Aged; Phosphopyruvate Hydratase; Synaptophysin; Urinary Bladder Neoplasms; Vimentin

Results of an immunohistochemical study in normal urothelium and transitional cell carcinomas of the bladder are presented. Paraffin-embedded material was confronted with immunoantisera against carcinoembryonic antigen (CEA), keratin (K), cytokeratin (CK) and epithelial membrane antigen (EMA). Immunohistochemical findings confirm the changes in reactivity of dysplastic urothelium and carcinoma in situ for CEA, CK and EMA, in comparison with normal urothelium. Statistically significant differences were also found, depending upon tumor stage, in staining of transitional cell carcinomas for K and CK. Expression of CK correlated with the tumor differentiation grade: normal urothelium and well-differentiated carcinomas showed a specific pattern of immunostaining for the basal cells, this pattern being lost in poorly differentiated carcinomas.

Topics: Adult; Aged; Antigens, Differentiation; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma in Situ; Carcinoma, Transitional Cell; Epithelium; Female; Humans; Keratins; Male; Membrane Glycoproteins; Middle Aged; Mucin-1; Neoplasm Staging; Urinary Bladder Neoplasms

Keratin No 17 expression (Mab E3) correlated with the stage and degree of malignancy of transitional-cell bladder carcinoma (TCC). Antibodies to keratin No 17 stained only the basal layer of the tumour growth in the TCC I while in TCC II-III positively stained cells were found everywhere in the tumour and staining was diffuse. Immunostaining for the simple keratins No 7, 8, 18, 19 in TCC I-III was diffuse. Distribution of keratin No 17 as well as that of the simple keratins was not regular in the metaplastic and anaplastic forms of TCC. Thus, the expression of keratin No 17 and simple keratins may serve an objective criterion of the tumour progression stage.

Topics: Carcinoma, Transitional Cell; Diagnosis, Differential; Humans; Immunohistochemistry; Keratins; Urinary Bladder Neoplasms

We examined the feasibility of performing non-radioactive in situ hybridization (ISH) in flow cytometrically sorted (tumor) cells with a chromosome #1 specific centromere probe. The study was performed in a model system of HL60 cells mixed with different quantities of HeLa cells. These latter cells were sorted directly onto poly-l-lysine coated glass slides on the basis of their keratin content, a cytoskeletal component not present in HL60 cells. Overall morphology of the separated HeLa cells was excellent and, after the ISH procedure, the appropriate number of ISH spots was observed in more than 85% of the sorted cells. This percentage did not differ significantly in cell mixtures with different percentages of HeLa cells (down to 1%). Sorting of HeLa cells in different phases of the cell cycle, and subsequent ISH, revealed the same spot number for chromosome #1 in all cell cycle stages, including mitosis. In the latter phase of the cell cycle we did not find a duplication of the chromosome #1 centromere, not even after sorting of the mitotic cells on the basis of specific labeling with an antibody to mitotin. The early G2 mitotin negative fraction, however, showed a significant percentage of cells with a duplicate spot number, most likely representing a tetraploid cell fraction in this HeLa cell culture. The protocol that evolved from these model studies was applied to cell suspensions of malignant body cavity effusions as well as solid bladder carcinomas. In several of these cases numerical chromosome aberrations could be detected by ISH more evidently after sorting on the basis of keratin labeling.

Topics: Antibodies, Monoclonal; Cell Cycle; Cell Cycle Proteins; Cell Separation; Centromere; Chromosomes, Human; Chromosomes, Human, Pair 1; DNA Probes; Flow Cytometry; HeLa Cells; Humans; Interphase; Keratins; Leukemia, Promyelocytic, Acute; Minichromosome Maintenance Complex Component 2; Neoplasms; Nuclear Proteins; Nucleic Acid Hybridization; Pleural Effusion; Tumor Cells, Cultured; Urinary Bladder Neoplasms

The clinical, microscopical, immunocytochemical and ultrastructural features of five cases of benign mesenchymal proliferative lesions of the urinary bladder, mimicking sarcoma, are presented. Four of the five patients are alive and disease-free following diagnosis, an interval ranging from 9 months to 9 years, mean 4 years. A fifth patient, who had a pseudosarcomatous stromal response adjacent to a urinary transitional cell carcinoma, now has invasive transitional cell carcinoma. The lesions revealed a striking microscopical, immunocytochemical and ultrastructural similarity to nodular fasciitis, suggesting the lesions represented a bizarre mesenchymal proliferative response to inflammation.

Topics: Adult; Carcinoma; Desmin; Female; Humans; Immunohistochemistry; Keratins; Lectins; Male; Microscopy, Electron; Middle Aged; Plant Lectins; Urinary Bladder Neoplasms; Vimentin

The main distinctive feature of carcinoma in schistosomal bladder is keratinized squamous cell carcinoma. Keratins/cytokeratins constitute a multigeneic family of structurally related polypeptide markers for the malignant state of epithelial cells. A monoclonal antibody (UNME/K1) regognizing keratins associated with squamous cell carcinoma of the human urinary bladder was generated at the Urology and Nephrology Center, Mansoura, Egypt (UNME), by fusion of spleenocytes from a BALB/c mouse immunized with a keratin extract (K1) of human squamous cell carcinoma and P3X63Ag8/U1 syngeneic myeloma cells. UNME/K1 was purified by a protein-A affinity column and was of the IgG2a type, as determined by immunoelectrophoresis and gel diffusion techniques. When tested against keratins of different types of urinary bladder tumors using enzym linked immunosorbent assay (ELISA), UNME/K1 reacted only with the high molecular weight keratin of squamous cell carcinoma and showed selectivity towards specific histopathological grades of tumors.

Topics: Animals; Antibodies, Monoclonal; Carcinoma, Squamous Cell; Humans; Immunoglobulin G; Keratins; Mice; Schistosomiasis haematobia; Urinary Bladder Neoplasms

Flow cytometry analysis of primary bladder carcinoma cells is appropriate for demonstrating the association between the presence of aneuploid cells and increased malignant potential for transitional cell carcinoma (TCC). Our laboratory previously has reported a specific alteration in the cytokeratin (CK) pattern of experimentally induced bladder carcinomas in rats. Unique immunohistochemically detectable changes in CK expression were the loss of CK 13 expression in invasive TCC cells and the loss of CK 19 expression in induced TCC. We now report the correlation of these changes in CK expression with cellular aneuploidy of the induced bladder carcinomas. Bladder carcinomas were induced in rats by direct instillation of N-methyl-N-nitrosourea. Immunohistochemistry was performed on the induced tumors using four commercially available monoclonal antibodies specific for CK 18 (TR1031), CK 13 (K8.12), CK 19 (K4.62), and the CKs 5, 7, and 8 (K8.13). Flow cytometry data from the induced bladder carcinomas was analyzed and the DNA index, proliferative index, and percent aneuploid cells were calculated for each time point. The percent aneuploid cells and CK 19 staining were tested statistically and were shown to be negatively correlated. We therefore hypothesize that the combination of the loss of CK 19 as detected by the antibody K4.62 coupled with the presence of aneuploid cells in histopathologically diagnosed invasive TCC is a significant factor in predicting the prognosis for any given diagnosis.

Topics: Animals; Carcinoma, Transitional Cell; Female; Flow Cytometry; Keratins; Mitosis; Neoplasm Invasiveness; Ploidies; Pregnancy; Rats; Rats, Inbred F344; Urinary Bladder Neoplasms

The new continuous cell line HOK-1 derived from a grade-III transitional-cell bladder carcinoma with foci of squamous and glandular differentiation was shown to retain this phenotypical heterogeneity for more than 45 passages in vitro. Electron microscopy revealed transitional as well as a considerable proportion of squamous carcinoma and adenocarcinoma cells. PAS-positive mucus was detected in numerous cells. These features were principally maintained when grown as multicellular spheroids and in nude mice. More pronounced signs of differentiation (i.e., expression of cytokeratins 10 and 11, formation of glandular structures) were found in xenograft tumours. Independently, cytokeratins 13, 18 and 19 were detected in vitro and in vivo, reflecting the urothelial origin. The line forms distinct colonies in soft agar, expresses Lewis-x and Lewis-y antigens and reacts with monoclonal antibodies (MAbs) against CEA, beta-HCG and URO-5. Cytogenetic analysis revealed several related clones with a rearrangement at chromosome 1 and loss of one X chromosome as common karyotypic changes in all clones. DNA content, as quantified by image analysis, showed a DNA stemline close to 2c. The new cell line HOK-1 can be used as an in vitro model to study the mechanisms of heterogeneous differentiation patterns in bladder cancer.

Topics: Aged; Biomarkers, Tumor; Blood Group Antigens; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cell Differentiation; DNA, Neoplasm; Female; HLA Antigens; Humans; Isoenzymes; Karyotyping; Keratins; Microscopy, Electron; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Cytokeratin shedding into urine was measured using a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) in 282 individuals. Samples included urine from normal controls, patients with urogenital conditions and bladder cancer patients. A monoclonal antibody prepared against cytokeratins extracted from a hyperkeratotic low grade squamous cell carcinoma (UNME/K1) was used in the assay. The results indicated reasonable levels of sensitivity (83%), specificity (67%) and overall accuracy (70%) in the detection of bladder cancer. The levels of sensitivity in detecting squamous and transitional cell carcinoma patients were 87 and 73% respectively. The low level of specificity was due to a high frequency of false positive results (55%) within the urogenital controls; this suggests that further immunochemical and immunohistopathological analyses of associated urothelial cytokeratins are required.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Schistosomiasis haematobia; Sensitivity and Specificity; Urinary Bladder Neoplasms

The L1 antigen was investigated as a marker of squamous differentiation in urothelium using a monoclonal antibody Mac387, and the results were compared with the expression of high molecular weight cytokeratins. L1 antigen was consistently demonstrated in all instances of partial and complete squamous metaplasia and in squamous carcinomas. In contrast, pure transitional cell carcinomas (except one with minor focal staining), adenocarcinomas and normal and hyperplastic urothelium did not label. In a few squamous carcinomas in situ, the pattern of labelling obtained with Mac387 was different from that seen in invasive squamous carcinomas and metaplasias. Compared with high molecular weight cytokeratins, the expression of L1 was more intense in areas of squamous differentiation. L1 expression, as identified by antibody Mac387, may therefore serve as a useful marker of squamous differentiation in urothelial lesions.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cell Adhesion Molecules, Neuronal; Cell Transformation, Neoplastic; Humans; Keratins; Leukocyte L1 Antigen Complex; Metaplasia; Urinary Bladder; Urinary Bladder Neoplasms

A mouse monoclonal antibody (CK1K10) specific for keratin was developed. Enzyme immunoassays (EIAs) were performed to detect keratin in urine samples using CK1K10 and anti-mouse IgG. A high degree of sensitivity and specificity was obtained in the detection of bladder cancer patients. A fast dot-ELISA was developed in which urine is applied onto nitrocellulose (NC) sheets or dipsticks. Dipsticks coated with urine samples can be stored at room temperature for more than 5 days and at -20 degrees C for more than 3 months without loss of efficiency. A high positive correlation was found between the fast dot-ELISA and other EIAs. The assay does not require sophisticated equipment nor highly trained technical staff and can be performed in less than 30 min. The keratin dot-ELISA could be a reliable screening test for the early detection of bladder cancer.

Topics: Animals; Antibodies, Monoclonal; Biomarkers, Tumor; Enzyme-Linked Immunosorbent Assay; Guinea Pigs; Humans; Immunoenzyme Techniques; Keratins; Mass Screening; Mice; Reagent Kits, Diagnostic; Urinary Bladder Neoplasms

Carcinosarcomas of the bladder are rare. We describe three such tumours, including an apparently unique case in which the components comprised liposarcoma and poorly-differentiated transitional cell elements. A second example included chondrosarcomatous elements. These two tumours showed architectural and immunocytochemical features which suggested that they had originated as carcinomas but had subsequently differentiated along both epithelial and mesenchymal pathways. The third tumour contained both carcinoma and osteosarcoma and may represent a collision tumour.

Topics: Aged; Aged, 80 and over; Carcinosarcoma; Female; Humans; Keratins; Male; S100 Proteins; Urinary Bladder Neoplasms

Twenty patients with bladder carcinoma (14 transitional cell carcinoma and six squamous cell carcinoma) and ten controls of (four with bilharzial lesions, three without and three with metaplasia and dysplasia) were subjected to immunocytochemical and immunohistochemical studies. Bladder and urine samples examination were carried out with the high molecular weight cytokeratin and broad spectrum keratin antibodies using peroxidase anti-peroxidase. Cytokeratins of high molecular weight were expressed in all squamous cancer while broad spectrum keratin was positive in 83.3%. The transitional cell carcinoma was positive in 50% with both cytokeratin and broad spectrum keratin. Urine shedded were positive in 83.3% and 66.7% of squamous cell carcinoma with high molecular weight cytokeratin and broad spectrum keratin respectively. However, each of them was positive in 50% of transitional carcinoma, and negative with non-malignant cases except with metaplasia and dysplasia.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Humans; Immunohistochemistry; Keratins; Schistosomiasis; Urinary Bladder Neoplasms

Using an explant tissue culture model developed by this group for use with human surgical and biopsy specimens, data are presented showing the response of normal and tumor bladder urothelium to radiation in combination with cis- and carboplatin. Cellular response is measured after two weeks in culture as a reduction in the extent of outgrowth from the explant. The outgrowth has been shown to be growing and to be epithelial. Results showed that when either drug or radiation is used singly, the tumour is resistant to treatment while the normal cells are severely affected. However, appropriate combinations of either drug with radiation reverse the unfavourable therapeutic ratio and result in higher tumour cell kill. The model may be useful for investigating mechanisms of radiation/chemotherapy action at the cellular level and, if integrated into appropriate clinical trials, may serve as an easy-to-use in vitro test for optimising single agent or combination therapy regimens.

Topics: Carboplatin; Cell Division; Cisplatin; Combined Modality Therapy; Culture Techniques; Drug Screening Assays, Antitumor; Humans; Keratins; Organoplatinum Compounds; Tumor Cells, Cultured; Urinary Bladder; Urinary Bladder Neoplasms

The discrimination of atypical (premalignant) cells from invasive neoplastic cells in primary bladder lesions is a major diagnostic problem in cytopathology and surgical pathology. We have used an animal model of urinary bladder carcinogenesis to determine the specific changes which occur in the expression of certain cytokeratins (CK) during the progression of lesions from regenerative hyperplasia and carcinoma in situ to transitional cell carcinomas. At sequential time points following exposure of the rat bladder epithelium to N-methyl-N-nitrosourea in vivo, immunohistochemical staining of CKs was evaluated in ethanol-fixed samples from the induced urothelial lesions using commercially available anti-CK mouse monoclonal antibodies. Specific changes were found in the expression of CKs 13, 18, and 19 during the neoplastic progression of induced urothelial lesions in the rat. These changes included the reciprocal loss of expression of CK 19 and the reappearance of CK 18 as malignant tumors developed. Invasive cells also did not express CK 13. Our results, based on the rat model, are similar to those reported by others on CK expression in human bladder tumors. Because these changes in CK expression occurred at specific points in the progression of urothelial lesions, the antibodies utilized in this study may be helpful in predicting the invasive potential of cells present in cytopathological specimens and tissue biopsies from human urothelial lesions.

Topics: Animals; Antibodies, Monoclonal; Blotting, Western; Epithelium; Female; Hyperplasia; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Methylnitrosourea; Rats; Rats, Inbred F344; Urinary Bladder; Urinary Bladder Neoplasms

These two cases of bladder tumours with an unusual histological appearance were observed at Hôpital Saint-Louis in 1988. They contained two cellular components: the usual epithelial type and a spindle cell type. Immunohistochemistry performed in order to identify the various cell contingents established the diagnoses of carcinosarcoma and spindle cell carcinoma and emphasised the differences and similarities between these two entities, which we believe can be differentiated.

Topics: Aged; Aged, 80 and over; Carcinoma; Carcinosarcoma; Desmin; Female; Humans; Immunohistochemistry; Keratins; Male; Myoglobin; Urinary Bladder Neoplasms; Vimentin

As bacillus Calmette-Guérin (BCG) immunotherapy is highly effective for most but not all superficial bladder tumors, there is a need to define predictors of response to this mode of treatment. We have investigated a panel of markers defined by monoclonal antibodies, directed against tumor-associated transitional cell carcinoma antigen (G4 and E7), epidermal growth factor receptor, cytokeratin (CK) 18 and blood group antigens A, B and H, using an indirect immunoperoxidase staining on paraffin sections. Twenty superficial bladder tumors (T1) treated with intravesical BCG therapy (10 responders and 10 nonresponders) were tested with this panel. Among the responders, expression of CK18 antigen was positive in 7 and negative in 3, whereas in the nonresponder group it was positive in 2 and negative in 8. The difference was statistically significant (p less than 0.05). Loss of expression of CK18 antigenicity was associated with recurrence or progression of superficial bladder tumors following BCG therapy, indicating that changes in CK patterns should be investigated as potential predictive markers for response to BCG.

Topics: Administration, Intravesical; Antibodies, Monoclonal; BCG Vaccine; Biomarkers, Tumor; Carcinoma, Transitional Cell; Humans; Immunoenzyme Techniques; Immunotherapy; Keratins; Urinary Bladder Neoplasms

Reactivity patterns of seven mouse monoclonal antibodies to human keratin 7 were compared using immunoblotting and immunohistochemistry on cultured cells and normal human and animal tissues. Differences in keratin specificities as determined by two-dimensional immunoblots and interspecies cross-reactivity data on 8 mammalian species suggest that at least six nonidentical epitopes of the keratin 7 molecule are recognized by this panel of reagents. Immunohistochemical examination of a panel of various human neoplasms with monoclonal antibodies monospecific for keratins 7, 18 and 19 revealed potential value of keratin subtyping in differential diagnosis of tumors in general and in subclassification of carcinomas in particular.

Topics: Animals; Antibodies, Monoclonal; Breast Neoplasms; Colonic Neoplasms; Cross Reactions; Fluorescent Antibody Technique; Humans; Intermediate Filament Proteins; Keratins; Neoplasms; Staining and Labeling; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Topics: Adenocarcinoma, Mucinous; Biomarkers, Tumor; Carcinoma, Transitional Cell; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Male; Microscopy, Electron; Middle Aged; Neoplasm Staging; Ureter; Ureteral Neoplasms; Urinary Bladder; Urinary Bladder Neoplasms; Vimentin

The purpose of this study was to determine the tissue distribution of certain cytokeratins in the urothelium. A series of ten cystectomy specimens containing normal tissues was investigated by immunocytochemical labeling with specific antibodies against cytokeratins 19, 18, 8, 7, 10. Monoclonal antibodies were used on frozen sections. Distribution of keratin was correlated with morphologic changes. All specimens were lined by a normal transitional epithelium. Low molecular weight cytokeratins (polypeptides 19, 18, 8, 7) were detected in all cell layers. Anticytokeratin 19 stained epithelium in a rather uniform way. Different antibodies specific for cytokeratin 18 reacted differently on the same tissue. With certain anticytokeratins 18 and 8, a higher staining intensity was found in superficial layers as compared with intermediate cell layers. So-called "umbrella" cells were strongly stained. Anticytokeratin 10 revealed very rare endocrine-like cells. This normal urothelial pattern is in agreement with previous reports. This pattern of expression of cytokeratin polypeptides differs from that of nonkeratinizing squamous epithelium. Therefore, histological differentiation of epithelia is accompanied by pronounced changes in the expression of cytokeratin polypeptides.

Topics: Carcinoma, Transitional Cell; Epithelium; Humans; Keratins; Urinary Bladder; Urinary Bladder Neoplasms

Normal epithelial and carcinoma cells of human bladder were investigated for the cytokeratin which is one of the intermediate filaments and comprises cytoskeleton using the immunofluorescence method. Carcinoma cell lines used were JTC-30, JTC-32, HUB-41 and T-24. In normal urothelium, keratin fibers were fine and straight with unchanged diameter and distributed regularly in the cytoplasm. By contrast, keratin fibers in bladder carcinoma cells were kinked and changed in diameter and were distributed irregularly in the cytoplasm. The above findings were most obvious in T-24 which formed undifferentiated carcinoma when transplanted to nude mice, and keratin fibers were dominantly located in the perinuclear area. The changes of keratin fibers appeared to be parallel to the grade of histological anaplasia of the tumor formed by implantation of bladder carcinoma cell line cells to nude mice. These observations suggest that the morphology of cytokeratin is a useful indicator for evaluating the grade of malignancy in transitional cell carcinoma.

Topics: Animals; Carcinoma, Transitional Cell; Epithelial Cells; Fluorescent Antibody Technique; Humans; Keratins; Mice; Neoplasm Staging; Neoplasm Transplantation; Tumor Cells, Cultured; Urinary Bladder; Urinary Bladder Neoplasms

The focus of my study was the immunohistological expressions of bladder tumour cells when stained by several antigens, those being; Carcinoembryonic antigen (CEA), Ferritin, Beta 2 Microglobulin (beta 2-MG), Keratin and Glycogen. There were 59 cases of bladder cancer studied using the Peroxidase-antiperoxidase technique (PAP) of staining. Specimens were formalin-fixed, paraffin embedded, and then sectioned. Some correlation was found between histological grade, stage and CEA incidence but was not found in the Ferritin or beta 2-MG. Keratin and Glycogen were detected in all cases. These staining patterns were mosaic according to higher grade and stage. It is believed this demonstrates that immunohistochemical expressions of staining with CEA, Keratin, and Glycogen may be a sensitive indicator of histological grade, stage.

Topics: Adult; Aged; Aged, 80 and over; beta 2-Microglobulin; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Transitional Cell; Female; Ferritins; Glycogen; Humans; Immunoenzyme Techniques; Immunohistochemistry; Keratins; Male; Middle Aged; Neoplasm Staging; Urinary Bladder Neoplasms

The focus of my study was the immunohistochemical studies of tumour cells induced by ADM intravesical instillation therapy when stained by several antigens, those being; Carcinoembryonic antigen (CEA), Ferritin, Beta 2 Microglobulin (beta 2-MG), Keratin and Glycogen before and after the therapy. Then I discussed as to their value in indicating therapeutic efficacy. There were 18 cases of bladder cancer studied using the Peroxidase-antiperoxidase technique (PAP) of staining. Data indicated a 44.4% response rate following therapy with intravesical ADM. Immunohistochemical evaluation showed improvement as indicated by not staining with CEA. Also observed was no change and slightly diffuse staining with Keratin: and a decrease in all layers of Glycogen expression: However this could not be observed with the Ferritin or beta 2-MG. It is believed this demonstrates that immunohistochemical expressions of staining with CEA, Keratin, and Glycogen may be a sensitive indicator of a therapeutic response to intravesical ADM.

Topics: Administration, Intravesical; beta 2-Microglobulin; Biomarkers, Tumor; Carcinoembryonic Antigen; Doxorubicin; Drug Evaluation; Female; Glycogen; Humans; Immunohistochemistry; Keratins; Male; Urinary Bladder Neoplasms

We report two cases of osteoclast-like giant cell tumour of urinary bladder associated with papillary transitional cell tumours. Both cases were morphologically identical to giant cell tumour of bone. The giant cells stained strongly for acid phosphatase which was resistant to tartrate digestion, a staining reaction typical of osteoclasts. In view of the ability of urinary bladder to induce metaplastic and neoplastic bone, we believe that these tumours may represent extraosseous giant cell tumours of bone.

Topics: Acid Phosphatase; Aged; Bone Neoplasms; Carcinoma; Carcinoma, Transitional Cell; Diagnosis, Differential; Giant Cell Tumors; Humans; Immunohistochemistry; Keratins; Male; Membrane Glycoproteins; Mucin-1; Muramidase; Osteoclasts; S100 Proteins; Urinary Bladder Neoplasms; Vimentin

The morphologic discrimination between low and high grade malignant tumor cells arising in the urinary bladder is a major diagnostic problem in cytopathology. Using immunochemical peroxidase staining of cytokeratins (CKs) of human bladder exfoliative cytology specimens, we have been able to discriminate between transitional cell carcinoma cells, atypical cells and normal bladder cells. Commercially available monoclonal antibodies used in this study were: anti-CK 13 (Sigma K8.12), anti-CK 5, 7 and 8 (Sigma K8.13), anti-CK 19 (Sigma K4.62), and anti-CK 18 (Transformation Res. 1091). When normal bladder cells are stained with these antibodies, CK 18 is specific for superficial cells; CK 13 is specific for the basal type cells and CKs 5, 7, 8 and 19 are expressed by all urothelial cell types. Four cases diagnosed by cytopathological criteria as 'atypical' and 7 diagnosed as 'positive' (various grades) were used in this study. Cytologic diagnosis was confirmed by histopathology in 7 cases. Tissue was not available for histopathology in 4 cases. Malignant cells with a 'basal' or 'immature' phenotype preferentially stained with CK 13 and were associated with increased metastatic or malignant potential. Patients with higher grade tumors had more cells positive for CK 13 than did patients with atypical or lower grade malignancies. Patients with well-differentiated, low grade tumors predominantly shed cells that expressed CK 18 and CK 19. High grade, invasive bladder lesions were characterized by many cells expressing CK 13, and fewer cells expressing CK 19. The combination of diagnosis by morphologic criteria on Papanicolaou-stained specimens with immunochemical characterization of the same cells for CKs facilitate an accurate diagnosis of bladder lesions.

Topics: Adult; Aged; Antibodies, Monoclonal; Carcinoma, Transitional Cell; Epithelium; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Urinary Bladder; Urinary Bladder Neoplasms

Cytokeratin 18 expression in transitional cells of 76 bladder tumors (11 grade 1, 33 grade 2 and 32 grade 3) and 10 normal biopsies was quantified by flow cytometry after immunolabeling with the anti-cytokeratin 18 monoclonal antibody RGE 53. Combined cytometric analysis of deoxyribonucleic acid content and cytokeratin 18 expression was conducted. Of 76 tumor biopsies 38 had a unimodal deoxyribonucleic acid profile with a deoxyribonucleic acid index close to 1.0, while the other 38 showed a bimodal profile: 1 peak with an index of 1.0 and a second peak with an index of greater than 1.0. The 11 grade 1 tumors belonged to the first group, whereas 10 of the 33 grade 2 and 28 of the 32 grade 3 tumors belonged to the second group. Antibody RGE 53 reacted with 4 per cent of the normal bladder cells (umbrella cells), 14 +/- 3 per cent of the cells from grade 1, 33 +/- 8 per cent from grade 2 and 56 +/- 10 per cent from grade 3 tumors. Analysis of cytokeratin 18 expression according to deoxyribonucleic acid content at the single cell level in tumors with a bimodal deoxyribonucleic acid profile showed that cytokeratin 18 was detected frequently in cells with a high deoxyribonucleic acid index but much less so in cells with an index of 1.0. Therefore, expression of cytokeratin 18 can be recognized as a marker of aggressiveness in bladder tumors, since it increases in parallel with tumor grade and cell deoxyribonucleic acid content.

Topics: Antibodies, Monoclonal; Carcinoma, Transitional Cell; Cell Separation; DNA, Neoplasm; Flow Cytometry; Humans; Keratins; Urinary Bladder; Urinary Bladder Neoplasms

Transitional carcinomas of the urinary bladder were examined immunohistochemically for keratin proteins with the use of polyclonal antiserum (TK, 41-65 kDa) and 3 monoclonal antibodies (KL 1, 55-57 kDa; PKK 1, nos. 19, 18, 8; and K 8.12, nos. 16, 13). Umbrella cells gave particularly strong staining for TK, KL 1 and PKK 1, whereas they were negative for K 8.12. Basal- and intermediate-layer cells in urothelial epithelium were moderately positive for all keratins. Brunn's nests cells showed comparatively slight or moderate keratin staining, and K 8.12 staining of Brunn's nests was higher than in urothelial epithelial cells. Transitional carcinoma (grades I and II) indicated uniform keratin distribution, and staining was strong with TK, while that of KL 1, PKK 1 and K 8.12 varied, and grade III tumors showed the lowest intensity of staining. K 8.12 staining in papillary transitional carcinomas was strongly positive in basal located tumor cells, as compared with apical tumor cells. Squamous cell carcinoma was varying positive to keratin reactions dependent on the degree of keratinization. Heterogenity of keratin distribution in papillary transitional carcinomas was given between basal tumor cells and well differentiated tumor cells including umbrella-like cells.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Epithelial Cells; Epithelium; Humans; Immunohistochemistry; Keratins; Urinary Bladder; Urinary Bladder Neoplasms

A case of atypical fibromyxoid tumor of the urinary bladder in a 32-year-old woman is reported. The patient had never complained of urinary symptoms, and bladder tumefaction was revealed fortuitously at pelvic ultrasound. Cystoscopy revealed a peanut-sized mass. Microscopically, the lesion was composed of strap- and tadpole-shaped cells resembling rhabdomyoblasts. For this reason, the tumor was initially diagnosed as embryonal rhabdomyosarcoma. However, immuno-histochemical study was negative for muscle origin, and the tumor has subsequently proved benign. The reported case illustrates the value of immuno-histochemical study in the evaluation of the true type of bizarre stroma cells in this pseudo-sarcomatous lesion. Their recognition is important, because the therapeutic consequences of misinterpreting this tumor as a sarcoma are great.

Topics: Adult; Female; Fibroma; Humans; Immunohistochemistry; Keratins; Urinary Bladder Neoplasms; Vimentin

Leukocyte infiltration has been studied with flow cytometry in a series of 76 bladder tumors. Cell DNA content and surface immunofluorescence with a panel of 6 monoclonal antibodies (Mabs) were evaluated using a double-staining technique. The following Mabs were used: anti-CD5 (ST1); anti-CD37 (SB3); anti-granulocytes (WEMG1); anti-monocytes/macrophages (PHM2); a Mab directed against a bladder tumor-associated antigen (G4), and the anti-keratin-18 RGE53. Antigens recognized by Mabs ST1, SB3 and WEMG1, were only expressed by cells with a DNA index close to 1, whatever the grade or the DNA profile of the tumor. Conversely, PHM2 was not only expressed by cells with a DNA index close to 1, but also by cells with a DNA index of more than 1.5, i.e., by cells of the second peak in tumors with a bimodal DNA profile. Thus the PHM2 epitope was not exclusively expressed by macrophages but also by urothelial tumor cells with an elevated DNA index. A significant positive correlation (r = 0.87; p less than 0.01) was found between DNA index and PHM2 reactivity among cells of the second peak in tumors with a bimodal DNA profile, whereas there was no correlation between DNA index and cytokeratin-18 expression and a weak negative correlation (r = -0.42; p less than 0.02) between DNA index and G4 Mab reactivity. These findings support the hypothesis of a cell fusion between human malignant urothelial cells and normal host macrophages.

Topics: Antibodies, Monoclonal; Antigens; Antigens, Neoplasm; DNA, Neoplasm; Flow Cytometry; Humans; Keratins; Leukocytes; Macrophages; Urinary Bladder Neoplasms

Two cases of signet-ring cell adenocarcinoma of the urinary bladder with a linitis plastica pattern of infiltration were studied. Mucin histochemistry indicated the presence of neutral and acid mucopolysaccharides and absence of sulphomucin. Immunocytochemistry of the tumour cells was positive for keratin and carcinoembryonic antigen, but negative for prostatic specific antigen and vimentin. The relevance of these observations to the differentiation and histogenesis of these tumours is discussed.

Topics: Adenocarcinoma, Mucinous; Adenocarcinoma, Scirrhous; Carcinoembryonic Antigen; Humans; Immunohistochemistry; Keratins; Linitis Plastica; Male; Middle Aged; Mucins; Staining and Labeling; Urinary Bladder; Urinary Bladder Neoplasms

A rat monoclonal antibody (MAb) termed RS-II (Ig M) was obtained by syngeneic immunization with rat bladder tumor cells induced by N-butyl, N-hydroxybutylnitrosamine (BBN). Immunocytochemical analysis showed that RS-II is also reactive with mouse, dog and human bladder tumor-cell lines and some other human tumor-cell lines but not myeloma or leukemia cells. Immunohistochemical examination of paraffin-embedded tissues has shown that RS-II is reactive with mouse, rat, dog and human bladder tumors (5/5, 5/5, 1/1, 31/49) and some other tumors, but not with normal human urothelium or normal rat tissues. The antigen is expressed on the majority of low-grade or well-differentiated tumors, but less on advanced, invasive tumors. The immunofluorescence staining pattern of cultured cells was cytoplasmic and filamentous. Immunoblotting revealed that the antigen is a Mr 54,000 to Mr 56,000 protein which was extracted with difficulty from the cultured cells with Triton X-100. The antigen was also detected in the culture supernatant by means of ELISA. Our results suggest that the epitope is expressed on cytoskeletons common to a wide variety of mammalian cells at a certain stage of differentiation.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Antigens, Neoplasm; Epitopes; Fluorescent Antibody Technique; Humans; Hybridomas; Immunoblotting; Keratins; Male; Rats; Rats, Inbred Strains; Tumor Cells, Cultured; Urinary Bladder Neoplasms

An 81-year-old woman was admitted to our clinic due to gross hematuria. A large bulky pedunculated mass was found in the bladder by cystoscopic examination. Subtotal cystectomy and bilateral cutaneostomy was performed on January 12, 1987. Histologically the tumor was composed of carcinomatous and sarcomatous elements. The carcinomatous element was composed fundamentally of grade 2, transitional cell carcinoma with numerous foci of squamous metaplasia. The sarcomatous element was composed of myxosarcomatous, chondro-sarcomatous pattern and non-differentiated malignant spindle cell component. Immunohistochemical examination demonstrated the presence of cytokeratin and epithelial membrane antigen in the spindle cell and more obvious carcinomatous regions, using the avidin-biotin conjugated immunoperoxidase technique. The patient died 3 months after operation. Autopsy findings showed multiple organ metastasis which were composed of carcinomatous and sarcomatous elements.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; Female; Humans; Keratins; Membrane Glycoproteins; Mucin-1; Neoplasms, Germ Cell and Embryonal; Urinary Bladder Neoplasms

Normal bladder urothelium and large spectrum bladder lesions have been investigated by immunohistochemistry with monoclonal antibodies of variable specificity (SK 56-23, a large spectrum antibody; SK 60-61, which reacts with cytokeratin polypeptides no. 8 and 18 of Moll's catalogue; SK 2-27, specific for polypeptides no. 14, 16 and 17). The normal urothelial pattern is in agreement with previous reports. In pathological conditions, modified immunostaining has been demonstrated in almost all cases. In detail, the cytoskeletal pattern detected in transitional cell papilloma seems to discriminate between types which are otherwise histologically similar. We also observed a correlation between higher degrees of malignancy and loss of specialization, as demonstrated by the increasing positivity for SK 60-61, which as a rule specifically stains "umbrella" cells, and SK 2-27, an antibody exclusively detected in cells of the basal layer. These findings indicate that the cytokeratin pattern may constitute a modern new tool for the pathologist in the diagnosis of urothelial proliferative disorders.

Topics: Antibodies, Monoclonal; Carcinoma, Transitional Cell; Cystitis; Epithelium; Histocytochemistry; Humans; Immunohistochemistry; Keratins; Metaplasia; Papilloma; Urinary Bladder; Urinary Bladder Diseases; Urinary Bladder Neoplasms

Cytokeratin shedding into the urine was measured using an enzyme-linked immunosorbent assay in 154 individuals. Samples include urines of normal controls, patients with urological conditions other than bladder cancer and bladder cancer patients. The assay results reflect the potential value of cytokeratin shedding in urine as a new urinary marker for bladder cancer. A 94% sensitivity for patients with squamous cell carcinoma of the bladder suggested the importance of using antibodies prepared against cytokeratin extracted from the same cell type of carcinoma as that to be detected. The relatively low sensitivity shown while detecting transitional cell carcinoma patients and the relatively low degree of assay specificity suggested the use of a panel of monoclonal antibodies specific to various types of cytokeratins of bladder carcinoma.

Topics: Adult; Antibodies, Monoclonal; Antibody Specificity; Biomarkers, Tumor; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratins; Male; Middle Aged; Predictive Value of Tests; Schistosomiasis haematobia; Sensitivity and Specificity; Urinary Bladder Neoplasms

A human ovarian Brenner tumor presenting a wide spectrum of benign and malignant histologic features was studied for its patterns of intermediate filament expression. All epithelial elements of the tumor, regardless of their morphologic type, contained cytokeratins as their only intermediate filament component. Differences were detected, however, between tumor nests that displayed transitional epithelium and those with squamoid features. These differences were manifested by the presence of cytokeratin 18, in the former type only, and by the abundance of cytokeratins 10/11 in the latter. We also detected mixed epithelial nests in which both features were present, suggesting that the transitional epithelium transforms in polar fashion into squamous epithelium. Examination of cytokeratin patterns found in urothelium and in the surface epithelium of the ovary pointed to certain differences from the Brenner tumor epithelia. The significance of these latter findings with regard to cellular transformation and histogenesis of the Brenner tumor are discussed.

Topics: Antibodies, Monoclonal; Brenner Tumor; Cytoskeletal Proteins; Desmin; Desmoplakins; Desmosomes; Epithelium; Female; Fluorescent Antibody Technique; Humans; Keratins; Membrane Glycoproteins; Middle Aged; Ovarian Neoplasms; Ovary; Urinary Bladder Neoplasms; Vimentin

The pattern of cytokeratins expressed in normal urothelium has been compared with that of various forms of transitional cell carcinomas (TCCs; 21 cases) and cultured bladder carcinoma cell lines, using immunolocalization and gel electrophoretic techniques. In normal urothelium, all simple-epithelium-type cytokeratins (polypeptides 7, 8, 18, 19) were detected in all cell layers, whereas antibodies to cytokeratins typical for stratified epithelia reacted with certain basal cells only or, in the case of cytokeratin 13, with cells of the basal and intermediate layers. This pattern was essentially maintained in low-grade (G1, G1/2) TCCs but was remarkably modified in G2 TCCs. In G3 TCCs simple-epithelial cytokeratins were predominant whereas the amounts of component 13 were greatly reduced. Squamous metaplasia was accompanied generally by increased or new expression of some stratified-epithelial cytokeratins. The cytokeratin patterns of cell culture lines RT-112 and RT-4 resembled those of G1 and G2 TCCs, whereas cell line T-24 was comparable to G3 carcinomas. The cell line EJ showed a markedly different pattern. The results indicate that, in the cell layers of the urothelium, the synthesis of stratification-related cytokeratins such as component 13 is inversely oriented compared with that in other stratified epithelia where these proteins are suprabasally expressed, that TCCs retain certain intrinsic cytoskeletal features of urothelium, and that different TCCs can be distinguished by their cytokeratin patterns. The potential value of these observations in histopathologic and cytologic diagnoses is discussed.

Topics: Antibodies, Monoclonal; Carcinoma; Carcinoma, Transitional Cell; Cell Differentiation; Epithelium; Fluorescent Antibody Technique; Humans; Keratins; Microscopy, Fluorescence; Reference Values; Tumor Cells, Cultured; Urinary Bladder Neoplasms; Urinary Tract; Urogenital Neoplasms

The level of urinary keratin was compared in patients with and without bladder cancer using the newly developed IRMAK-18. Patients with bladder cancer had higher levels of keratin (74.6 +/- 146.9) than normals (2.5 +/- 4.7). When the upper limit of normal was set at keratin less than 20 ng./ml., the test was highly specific (98%) for bladder cancer but had a sensitivity of only 46%. Adjusting to keratin less than 10 ng./ml. increased the sensitivity to 57% but decreased the specificity to 91%. Elevated keratins were associated with stage and grade, aneuploidy and disease history. The IRMAK-18 assay for keratin may be useful for monitoring the clinical progress of some patients.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Transitional Cell; DNA; Flow Cytometry; Humans; Immunoassay; Immunoenzyme Techniques; Keratins; Middle Aged; Radiometry; Urinary Bladder Neoplasms

Topics: Antibodies, Monoclonal; Carcinoma, Transitional Cell; Flow Cytometry; Humans; Keratins; Urinary Bladder Neoplasms

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Transitional Cell; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Urinary Bladder Neoplasms

Expression of low and high molecular weight cytokeratin proteins was investigated immunohistochemically in a variety of transitional and squamous epithelial lesions of the urinary tract with and without schistosomiasis. The monoclonal antibodies used were CAM 5.2 and NCL5D3 for low, PK 63 and 121 for high, and MAK 6 for a broad range of intermediate molecular weight cytokeratins. On staining with CAM 5.2 and NCL5D3, urothelial hyperplasias (n = 12) and grades 1 (n = 5) and 2 (n = 10) papillary transitional cell carcinomas showed labelling patterns quite distinct from carcinoma in situ (n = 4) and non-papillary grades 2 (n = 6) and 3 tumours (n = 3). Among squamous lesions only focal positivity was obtained in 14 of 22 moderate to poorly differentiated squamous cell carcinomas. By contrast, PK 63 and 121 stained squamous lesions exclusively. MAK 6 stained the whole range of urothelial and squamous lesions with the exception of squamous metaplasias. Polyclonal antikeratin adequately labelled spindle cell areas of high grade tumours. The distinctive staining patterns given by these or similar antibodies may help in the identification of squamous metaplasia and in diagnosing tumours of the urothelium.

Topics: Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Epithelium; Humans; Keratins; Lymphatic Metastasis; Molecular Weight; Schistosomiasis haematobia; Urinary Bladder Neoplasms

The clinicopathologic and immunohistochemical features of 16 pure adenocarcinomas primary in the urinary bladder were reviewed. Only 3 patients were found to have disease confined to the urinary bladder. Of 13 cases with follow-up only 3 are free of disease. Histologically, the tumors were classified as signet ring cell (3), colloid (3), colonic type (5), clear cell (1), and not otherwise specified (NOS, 4). Immunohistochemically, all tumors but one colloid carcinoma were immunoreactive for cytokeratin and epithelial membrane antigen, and most tumors were likewise immunoreactive for carcinoembryonic antigen. Eight cases were immunoreactive for Leu M1 antigen. Prostate specific antigen, S-100 protein, and placental alkaline phosphatase were uniformly negative. No correlation between immunohistochemical profile and histologic type or clinical outcome was found. The utility of immunohistochemistry and other pathologic findings is reviewed.

Topics: Adenocarcinoma; Adenocarcinoma, Mucinous; Adult; Aged; Carcinoembryonic Antigen; Female; Humans; Keratins; Male; Membrane Proteins; Middle Aged; Mucin-1; Urinary Bladder; Urinary Bladder Neoplasms

Thirty-seven transitional-cell carcinomas (TCC) of the urinary bladder were analyzed by DNA flow cytometry (FCM). After labelling of the cell suspensions with antibodies to cytokeratin, the cytokeratin-positive cells and the non-epithelial cytokeratin-negative cells could be analyzed separately. After estimation of S- and G2M phase, 3/17 cases (18%) with a normal DNA index showed elevated proliferative levels, among cytokeratin-labelled suspensions only. Of these 17 cases, 14 showed chromosomal abnormalities. The remaining 20 cases were abnormal, irrespective of the technique used. Although immuno-labeling of tumor cells for cytokeratin in FCM increases the sensitivity of this method in detecting aneuploid tumors or tumors with high proliferation fractions, the discriminating power of chromosomal analysis of TCC is greater than FCM.

Topics: Carcinoma, Transitional Cell; Cell Cycle; Chromosome Aberrations; DNA, Neoplasm; Female; Flow Cytometry; Humans; Karyotyping; Keratins; Male; Urinary Bladder Neoplasms

Tissue polypeptide antigen (TPA) both in normal and neoplastic urinary bladders has been studied by immunocytochemistry. A comparison of TPA with epithelial membrane antigen (EMA), keratins and carcinoembryonic antigen (CEA) in various tumor grades and stages has been performed better to define TPA role in bladder carcinomas. Well-differentiated tumors were strongly stained for TPA with a uniform staining intensity. Undifferentiated tumors were weakly stained for TPA with an uneven staining intensity. There was no relation between TPA findings and stages of invasion. However, TPA seems to be a very helpful diagnostic tool for tumor grading and staging.

Topics: Adult; Aged; Antigens, Neoplasm; Biopsy; Carcinoma, Transitional Cell; Epithelium; Humans; Keratins; Middle Aged; Neoplasm Staging; Peptides; Prognosis; Staining and Labeling; Tissue Polypeptide Antigen; Urinary Bladder; Urinary Bladder Neoplasms

Topics: Adenocarcinoma; Breast Neoplasms; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Cytoskeleton; Diagnosis, Differential; Epithelium; Female; Gastrointestinal Neoplasms; Humans; Intermediate Filament Proteins; Intermediate Filaments; Keratins; Lung Neoplasms; Neoplasms; Skin Neoplasms; Urinary Bladder Neoplasms; Uterine Cervical Neoplasms

Squamous cell carcinomas of the urinary bladder and the epithelial lesions associated with infection by Schistosoma haematobium were histopathologically and immunohistochemically described for keratin proteins (TK, 41-65 kDa; KL1, 55-57 kDa; PKK1, 40, 45 and 52.5 kDa), involucrin, and epithelial membrane antigen (EMA). Normal urothelial epithelium was positive for all keratins, and showed absent or slight reactions for involucrin and EMA in superficial umbrella cells. The intestinal type of epithelium was composed of columnar cells and small basal cells; TK was positive in the basal cells, KL1 staining was positive in the columnar cells, whereas PKK1 was negative or slight in the columnar cells. Involucrin was confined to columnar cells. Squamous metaplastic epithelium showed a rather regional keratin distribution: TK was distributed in all layers, KL1 decorated upper spinous and granular layers, but PKK1 did not bind, and involucrin staining existed only in upper spinous and granular cells. Keratin expression in squamous cell carcinomas indicated heterogeneity and its stainability was dependent on the degree of keratinization: The G 1 type revealed strong reaction, the G 2 type showed a similar distribution pattern, but the staining intensity was less, and the G3 type showed irregular staining with decreased intensity. Involucrin staining was limited to keratinized cells of carcinoma as was that for EMA.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Female; Histocytochemistry; Humans; Immunochemistry; Keratins; Male; Membrane Proteins; Middle Aged; Mucin-1; Protein Precursors; Schistosomiasis haematobia; Urinary Bladder Neoplasms

The distribution of tissue polypeptide antigen (TPA), epithelial membrane antigen (EMA) and various cytokeratins was studied with immunohistochemical techniques in malignant, normal and inflamed urothelium. TPA and cytokeratins 8, 18 and 19 were similarly distributed and there was no difference between benign and malignant conditions. All keratins were most abundant in the outer cell layers of the urothelium. EMA was present primarily in the so called umbrella cells. Increased levels of TPA in urine and serum are found in patients with urothelial carcinoma. TPA is reported to be related to the cytoskeletal keratins and its presence definitely does not imply malignant transformation as we have demonstrated it in both benign and malignant urothelium. The increased levels of TPA in urine and serum are probably due to a more rapid turnover and autolysis of cells and may parallel a more aggressive tumor behaviour.

Topics: Antibodies; Cystitis; Humans; Immunohistochemistry; Keratins; Membrane Glycoproteins; Mucin-1; Peptides; Tissue Polypeptide Antigen; Urinary Bladder; Urinary Bladder Neoplasms

The monoclonal antibody CAM 5.2 was used to investigate the expression of lower molecular weight cytokeratin proteins immunocytochemically in conventionally processed samples of schistosoma-associated squamous metaplasias and carcinomas of the urinary bladder. The antibody did not differentiate between squamous metaplasias and well differentiated squamous carcinomas, though it did focally label moderate to poorly differentiated tumours. The results suggest that squamous carcinomas of the bladder in this setting differ fundamentally from such tumours at other sites in their patterns of cytokeratin protein expression.

Topics: Antibodies, Monoclonal; Carcinoma, Squamous Cell; Humans; Keratins; Metaplasia; Molecular Weight; Schistosomiasis haematobia; Urinary Bladder; Urinary Bladder Neoplasms

A rapidly fatal bladder tumour which had the features of a rhabdoid tumour was studied by sequential biopsies and at autopsy. This is the first rhabdoid tumour recorded at this site and the first in which there was co-existent transitional cell carcinoma. The possibility that rhabdoid tumour is histogenetically heterogeneous is discussed.

Topics: Carcinoma, Transitional Cell; Female; Humans; Immunohistochemistry; Keratins; Microscopy, Electron; Middle Aged; Neoplasms, Multiple Primary; Phosphopyruvate Hydratase; Rhabdomyosarcoma; Urinary Bladder Neoplasms; Vimentin

Topics: Acridine Orange; Antibodies, Monoclonal; Flow Cytometry; Histocytochemistry; Humans; Immunologic Techniques; Keratins; Neoplasm Invasiveness; Specimen Handling; Urinary Bladder; Urinary Bladder Neoplasms

The flow cytometric (FCM) analysis of carcinomas is often hampered by the presence of stromal and inflammatory cells in the cell suspensions obtained from such neoplasms. Therefore, an FCM method was developed to distinguish epithelial from nonepithelial cells by using polyclonal and monoclonal antibodies to (cyto)keratins, the epithelial type of intermediate filament proteins. Using a model system of cultured bladder carcinoma (T24) and leukemia (MOLT-4) cells, we tested our hypothesis and procedures by labeling cell mixtures with these antibodies. After incubation with an appropriate intermediate filament antibody and propidium iodide staining, the DNA content and distribution of T24 cells could be analyzed separately from MOLT-4 cells. When applied to cell suspensions of endometrial carcinomas, bladder carcinomas and Grawitz tumors, only the epithelial (primarily carcinoma) cells were stained for cytokeratin; these cells could thus be analyzed separately from stromal, inflammatory and other nonepithelial cells. In this way, a more accurate FCM analysis of the malignant fraction within a tumor can be achieved.

Topics: Adenocarcinoma; Carcinoma, Transitional Cell; Cell Cycle; Cells, Cultured; DNA, Neoplasm; Epithelium; Flow Cytometry; Humans; Immunologic Techniques; Intermediate Filament Proteins; Keratins; Urinary Bladder Neoplasms; Vimentin

Keratin was identified with the aid of polyclonal antisera in the cytoplasm in over 90% of the transitional cell carcinomas investigated. The intensity of staining increased with the degree of dedifferentiation. Detection of cytokeratin with monoclonal antibodies was successful in over 80% of samples. All squamous cell carcinomas of the bladder were strongly positive for keratin and cytokeratin. CEA was found in 20% of the G1 and 40% of the G2 and G3 carcinomas of the bladder. Both the prostatic epithelium markers PSA and PAP and the monoclonal antibody Ca1 were negative in all cases.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Humans; Keratins; Urinary Bladder Neoplasms

Thirty-eight transitional-cell carcinomas (TCC) were analyzed by flow cytometry (FCM) using propidium iodide for DNA analysis and antibodies to cytokeratin by indirect immunofluorescence. By means of two-dimensional FCM analysis, cytokeratin-positive tumor cells could be analysed separately from cytokeratin-negative stromal and inflammatory cells. This resulted in an 18% increase in sensitivity of FCM detection of aneuploidy (10/38 samples with one-parameter DNA analysis versus 15/38 samples with two-parameter DNA and cytokeratin analysis). In addition, S-phase could be determined in the 15 aneuploid samples by means of two-parameter analysis where this was not possible using only DNA content because of the overlap of diploid and aneuploid populations. FCM analysis allowed quantification of the percentage of tumor cells expressing cytokeratin 18 which has previously been shown to correlate quantitatively with higher grade, higher stage TCC. The quantitative measurement of tumor-cell expression of cytokeratin 18 by FCM analysis appears to provide additional information of potential prognostic value, independent of tumor-cell ploidy and proliferative fractions.

Topics: DNA; Flow Cytometry; Fluorescent Antibody Technique; Humans; Interphase; Keratins; Prognosis; Propidium; Urinary Bladder; Urinary Bladder Neoplasms

Three hundred rat bladders bearing tumors induced by N-butyl-N-4-(OH)butyl-nitrosamine (BBN) were examined by routine histologic study and immunohistochemical staining of intermediate filament types. Smaller lesions were similar to human urothelial dysplasia histologically and immunohistochemically. Progression of the lesions demonstrated large exophytic papillomas with extensive endophytic epithelial growth into abundant stroma. These lesions showed increasing predominance of squamous over transitional elements. Immunohistochemical findings confirmed these results and also demonstrated that morphologically indifferent cells, even in early lesions, express heavier cytokeratins characteristic of keratinizing squamous epithelium. These results demonstrate that BBN-induced bladder tumors show marked quantitative and qualitative differences from the most common, purely transitional, human bladder carcinomas. However, the development in BBN-treated rat bladders of two tumor types, squamous and transitional, from an altered urothelium may serve as an attractive model for further study of the molecular genetics of keratin expression.

Topics: Animals; Antibodies, Monoclonal; Butylhydroxybutylnitrosamine; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Epithelium; Fluorescent Antibody Technique; Histocytochemistry; Keratins; Male; Papilloma; Rats; Rats, Inbred ACI; Urinary Bladder Neoplasms; Vimentin

Histogenesis of squamous cell carcinoma in two prostates heavily affected by schistosomiasis was determined immunohistochemically by localization of two prostatic specific markers and keratin. The demonstration of prostatic specific antigen and keratin served to differentiate between metaplasia and squamous cell carcinoma associated with prostatic schistosomiasis from other prostatic and urinary bladder neoplasms.

Topics: Acid Phosphatase; Antigens, Neoplasm; Carcinoma, Squamous Cell; Diagnosis, Differential; Humans; Keratins; Male; Neoplasm Metastasis; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms; Schistosomiasis; Seminal Vesicles; Urinary Bladder Neoplasms

Antibodies to intermediate filament proteins were used to study different cell layers in normal human transitional epithelium, 16 human transitional cell carcinomas, and two cell lines derived from human bladder carcinomas. Conventional rabbit antisera to human skin keratins stained all layers of the transitional epithelium from bladder, ureter, and kidney. A slightly higher staining intensity was found in the basal and superficial layers as compared with the intermediate cell layers. A monoclonal antibody to cytokeratin 18 (RGE 53), however, stained only the superficial cell layer of transitional epithelium, the so-called umbrella cells. In well-differentiated (grade I) transitional cell carcinomas, RGE 53 stained only the superficial cells of papillary structures. In higher grade papillary tumors, RGE 53 also stained cells within the basal and intermediate layers, whereas in high-grade, invasive tumors almost all tumor cells were RGE 53 positive. These results show that monoclonal antibodies to cytokeratins can provide both an indication of processes involved in neoplastic progression of bladder tumors and a means of studying the molecular relationship of the tumor cells to normal cells.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Antigen-Antibody Reactions; Carcinoma, Transitional Cell; Cell Transformation, Neoplastic; Epithelium; Histocytochemistry; Humans; Immune Sera; Intermediate Filament Proteins; Keratins; Rabbits; Urinary Bladder Neoplasms

Monoclonal antibodies were generated by immunizing rats with mouse bladder carcinomas, fusing their spleen cells with NS-1 myeloma cells and selecting for antibodies that bound to mouse bladder carcinomas. One of the antibodies, IG5, recognizes an antigenic determinant which is present in mouse bladder carcinoma cytoskeletons and is not detectable in the normal bladder epithelium. Indirect immunofluorescence studies revealed that the antigen is expressed intracellularly and is organized in the form of filamentous arrays. The antigen was detected by peroxidase--antiperoxidase immunocytochemistry in stratified epithelia and glands derived from these, but has not been observed in any tissues of mesenchymal or neuronal origin. Various normal and neoplastic human tissues were subsequently tested for reactivity with antibody IG5. Antigen expression in normal tissues was similar to that in the mouse. Most carcinomas of the bladder and lung were stained, while all of eleven colon carcinomas were negative. Antibody IG5 immunoprecipitated radio-iodinated peptides of 58, 56, 52 and 43 kD molecular weight from mouse bladder carcinomas. Western blotting experiments with antibody IG5 demonstrated bands of 56 and 50 kD in a keratin-enriched fraction of the bladder carcinoma cytoskeleton. Antibody IG5 reacted with molecules which have several properties typical of cytoskeletal keratin peptides. Our findings are discussed in the context of previously described keratin peptides and relevant monoclonal antibodies.

Topics: Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Cytoskeleton; Epidermis; Epithelium; Epitopes; Fluorescent Antibody Technique; Hybridomas; Immunoenzyme Techniques; Keratins; Mice; Mice, Inbred BALB C; Precipitin Tests; Urinary Bladder Neoplasms

Intermediate filament proteins of normal epithelia of the human and the bovine male urogenital tract and of certain human renal and bladder carcinomas have been studied by immunofluorescence microscopy and by two-dimensional gel electrophoresis of cytoskeletal fractions from microdissected tissue samples. The patterns of expression of cytokeratin polypeptides differ in the various epithelia. Filaments of a cytokeratin nature have been identified in all true epithelial cells of the male urogenital tract, including renal tubules and rete testis. Simple epithelia of renal tubules and collecting ducts of kidney, as well as rete testis, express only cytokeratin polypeptides nos. 7, 8, 18, and 19. In contrast, the transitional epithelia of renal pelvis, ureter, bladder, and proximal urethra contain, in addition to those polypeptides, cytokeratin no. 13 and small amounts of nos. 4 and 5. Most epithelia lining the human male reproductive tract, including those in the epididymis, ductus deferens, prostate gland, and seminal vesicle, synthesize cytokeratin no. 5 in addition to cytokeratins nos. 7, 8, 18, and 19 (cytokeratin no. 7 had not been detected in the prostate gland). Cytokeratin no. 17 has also been identified, but in very low amounts, in seminal vesicle and epididymis. The cytokeratin patterns of the urethra correspond to the gradual transition of the pseudostratified epithelium of the pars spongiosa (cytokeratins nos. 4, 5, 6, 13, 14, 15, and 19) to the stratified squamous epithelium of the fossa navicularis (cytokeratins nos. 5, 6, 10/11, 13, 15, and 19, and minor amounts of nos. 1 and 14). The noncornified stratified squamous epithelium of the glans penis synthesizes cytokeratin nos. 1, 5, 6, 10/11, 13, 14, 15, and 19. In immunofluorescence microscopy, selective cytokeratin antibodies reveal differential staining of different groups or layers of cells in several epithelia that may relate to the specific expression of cytokeratin polypeptides. Human renal cell carcinomas show a simple cytokeratin pattern consisting of cytokeratins nos. 8, 18, and 19, whereas transitional cell carcinomas of the bladder reveal additional cytokeratins such as nos. 5, 7, 13, and 17 in various proportions. The results shows that the wide spectrum of histological differentiation of the diverse epithelia present in the male urogenital tract is accompanied by pronounced changes in the expression of cytokeratin polypeptides and suggest that tumors from different regions of the ur

Topics: Animals; Cattle; Cytoskeletal Proteins; Electrophoresis, Agar Gel; Epithelial Cells; Epithelium; Fluorescent Antibody Technique; Genitalia, Male; Humans; Intermediate Filament Proteins; Keratins; Kidney Neoplasms; Male; Microscopy, Fluorescence; Peptides; Urinary Bladder Neoplasms; Urinary Tract

Keratin was found in more than 90% of transitional cell carcinomas of the bladder in the cytoplasma with polyclonal antibodies. Intensity increased with dedifferentiation. Cytokeratin was detected with monoclonal antibodies in more than 80%. Squamous cell carcinoma of the urinary bladder was always strongly positive for keratin and cytokeratin. CEA was found in 20% of G1 and 40% of G2 and G3 carcinomas of the urinary bladder. The prostatic epithelium markers PSA and PAP were always negative also Ca1.

Topics: Acid Phosphatase; Antibodies; Antigens, Neoplasm; Carcinoembryonic Antigen; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Humans; Immunoenzyme Techniques; Keratins; Male; Prostate; Prostate-Specific Antigen; Urinary Bladder Neoplasms

Carcinomas of different origin have been tested in immunofluorescence microscopy with the monoclonal murine antibodies CK1-CK4, which recognize a single cytokeratin polypeptide (human cytokeratin No. 18) present in simple but not in stratified squamous epithelia, and with the monoclonal antibody KG8.13 and guinea pig kerA antibodies, both of which recognize a variety of cytokeratins common to almost all epithelial cell types. Tumors derived from simple epithelia, including adenocarcinomas and some other tumors such as ductal breast carcinomas, were strongly stained by all three antibodies. So was a transitional carcinoma of the bladder. In contrast, basal cell epithelioma, cloacogenic carcinoma, and squamous cell carcinoma of skin, tongue, and esophagus appeared negative with CK1-CK4 but positive with the other two antibodies. Other squamous cell carcinomas derived from epiglottis and cervix uteri showed a mixture of positive and negative cells when tested with CK1-CK4, although all tumor cells were positive when tested with KG8.13 and with kerA. Thus, use of an appropriate collection of cytokeratin antibodies with different specificities not only allows tumors of epithelial origin to be distinguished from other tumor types but, in addition, allows a further subdivision of carcinomas in relation to their histologic origin.

Topics: Adenocarcinoma; Animals; Antibodies, Monoclonal; Antibody Specificity; Carcinoma; Carcinoma, Transitional Cell; Colonic Neoplasms; Fluorescent Antibody Technique; Guinea Pigs; Histocytochemistry; Humans; Intermediate Filament Proteins; Keratins; Mice; Urinary Bladder Neoplasms

An antigen, termed "urothelium membrane antigen" (UMA), confined to urothelium and most abundantly associated with the asymmetric unit membrane of the terminally differentiated luminal cells was identified with a monospecific (absorbed or affinity purified) rabbit antiserum obtained by immunization with membranes from normal bovine urothelium. A cross-species immunofluorescence-positive reaction was observed in normal bladder: Luminal urothelial membrane reacted most intensely. Underlying normal urothelial cells also showed a weaker cytoplasmic reaction. Specific luminal membrane labeling was confirmed by transmission and scanning electron microscopy. Specificity of the anti-UMA differed from the general epithelial reactivity obtained with antibodies to bovine urothelium cytosol. Anti-UMA did not cross-react with keratin, although there was some evidence of a filamentous localization in the cytoplasm of permeabilized cells. UMA was variably expressed by urothelial carcinoma-derived cell lines to a degree apparently related to the degree of cell differentiation, showing the highest positivity on the well-differentiated RT4 and RT112 lines and only a very weak reaction with the anaplastic MGHU-1 (EJ) and T24 lines. On immunoblots, anti-UMA reacted with a peptide of approximately 54,000 daltons.

Topics: Animals; Antigens, Surface; Cattle; Cell Differentiation; Cell Line; Cross Reactions; Epithelium; Fluorescent Antibody Technique; Keratins; Microscopy, Electron, Scanning; Species Specificity; Urinary Bladder; Urinary Bladder Neoplasms

Using two human tumour cell lines, T24 bladder carcinoma and Molt-4 leukemia, flow-cytometric DNA analysis of pure and mixed cell populations was performed using cellular cytokeratin content to distinguish cytokeratin-containing carcinoma cells from leukemia cells which do not contain cytokeratin. Using cytokeratin content to gate DNA analysis, the same specificity and sensitivity of cellular DNA content and distribution measurement could be achieved by single-pass FCM analysis of a mixture of the two cell types as was seen when analysing pure populations of the two cell lines. This technique has broad applicability to FCM analysis of mixed populations composed of cells from different tissues of origin.

Topics: Antibodies; Carcinoma, Transitional Cell; Cell Line; Cell Separation; Cytoskeleton; DNA, Neoplasm; Flow Cytometry; Humans; Keratins; Leukemia, Lymphoid; Urinary Bladder Neoplasms

Monoclonal antibodies were raised against the detergent-insoluble cytoskeletal fraction of the human bladder carcinoma cell line, EJ. By immunofluorescence, one of these antibodies, H10-1, localized to keratin filaments in EJ and three out of five other transitional-cell carcinoma lines. Primary normal-human urothelial cells in culture were not recognized by H10-1. Strong staining of keratin filaments was also seen in 11 human adenocarcinoma cell lines, but it was only seen in a subpopulation of cells in one out of four squamous-cell carcinoma lines and not at all in two normal diploid human epidermal keratinocyte strains. Immunofluorescence on frozen sections of mouse, human, and rabbit tissues showed that H10-1 recognized the upper layer of the transitional epithelium of mouse and rabbit bladder, and a subset of simple epithelia (mouse stomach glands and some colon mucosal glands, but not kidney tubules, small intestine, ovary germinal epithelium, or endometrium; some human colon mucosal glands and glandular breast epithelium, but not endometrium). The antigen which is recognized by this antibody was not detected in examples of stratified squamous epithelium (mouse skin, nonglandular stomach, or esophagus; human skin, esophagus, or rectum) or nonepithelial tissue. On frozen sections of human tumors, the H10-1 antibody recognized 9 out of 14 colon carcinomas and 4 out of 6 breast carcinomas but did not recognize two sarcomas or a melanoma. This antibody apparently does not recognize its antigen in the presence of sodium dodecyl sulfate (SDS) because brief exposure of methanol-fixed cells to 0.1% SDS reversibly inhibited the binding of the H10-1 monoclonal antibody to keratin filaments by immunofluorescence, even when the binding of a rabbit polyclonal antikeratin antibody was unaffected. Two-dimensional gel-electrophoretic analysis of keratin-enriched fractions which had been isolated from various cell lines disclosed a good correlation between the presence of a 47-kdalton, pI-5.35 polypeptide and positive immunofluorescence with this antibody. These observations suggest that a certain keratin or keratin-associated protein is specifically expressed in a subpopulation of transitional and simple epithelial cells, and cultured cells derived from these epithelia. The monoclonal antibody described here may therefore be useful for increasing the precision of histological classification of epithelia, as well as the verification of the origin of certain cultured

Topics: Animals; Antibodies, Monoclonal; Antigens; Carcinoma, Transitional Cell; Cell Line; Epithelium; Fluorescent Antibody Technique; Humans; Keratins; Mice; Urinary Bladder; Urinary Bladder Neoplasms

Immunoperoxidase staining for human keratin proteins (hKP) was performed cytochemically in samples from 79 cancer patients, and histochemically in samples from 134 cancer patients. Immunohistochemically, hKP was present in almost all patients with squamous cell carcinoma (lung), transitional cell carcinoma (urinary bladder), adenocarcinoma (lung, stomach, breast, ovary), bronchiole-alveolar carcinoma (lung), and large cell carcinoma (lung). It was detected in 40% of the patients with small cell carcinoma (lung). Histochemically, hKP was present in patients with squamous cell carcinoma (lung, uterine body), adenocarcinoma (lung, stomach, colon, gall bladder, thyroid, uterine body), adenosquamous cell carcinoma (gall bladder, uterine body), Signet-ring cell carcinoma (stomach), clear cell carcinoma (uterine body) and undifferentiated carcinoma (uterine body). However, it was not detected in patients with brochiole ++-alveola ++ carcinoma (lung) and mucinous carcinoma (gall bladder).

Topics: Breast Neoplasms; Colonic Neoplasms; Female; Histocytochemistry; Humans; Keratins; Lung Neoplasms; Male; Neoplasms; Stomach Neoplasms; Urinary Bladder Neoplasms; Uterine Neoplasms

Topics: Animals; Cell Transformation, Neoplastic; Etretinate; Female; Humans; Keratins; Male; Middle Aged; Neoplasm Recurrence, Local; Papilloma; Skin Neoplasms; Tretinoin; Urinary Bladder Neoplasms

Human epithelial cells contain, intermediate-sized filaments formed by polypeptides related to epidermal alpha-keratin ("cytokeratins") which are expressed in different combinations in different epithelia. Using cytoskeletal proteins from human biopsies and autopsies we have examined, by two-dimensional gel electrophoresis and immunoblotting experiments, the cytokeratin polypeptide patterns of diverse primary and metastatic carcinomas and have compared them with those of corresponding normal epithelial tissues and cultured cells. Five groups of carcinoma cytokeratin patterns can be discriminated. (1) Cytokeratins typical of simple epithelia (polypeptides Nos. 7, 8, 18, 19) are expressed, in various combinations, by many adenocarcinomas, for example those of gastrointestinal tract. (2) Cytokeratins typical of stratified epithelia (Nos. 1, 5, 6, 10, 11, 14-17) are found, in various combinations, in squamous cell carcinomas of skin and tongue. (3) Complex patterns showing polypeptides Nos. 7, 8, 18, 19, and one basic component (No. 5 or 6) are detected in certain carcinomas of the respiratory tract and the breast. (4) Complex patterns containing cytokeratins widespread in stratified epithelia (Nos. 4-6, 14-17) as well as components Nos. 8 and 19 occur in diverse squamous cell carcinomas derived from non-cornified stratified epithelia, with or without additional small amounts of cytokeratin No. 18. (5) Patterns of unusually high complexity can be found in some rare tumors as is shown for a cloacogenic carcinoma. No significant qualitative changes of expression of cytokeratins were found when primary tumors and metastases were compared. When compared with cytokeratin patterns of normal epithelia, carcinomas of the first type usually display a high degree of relatedness to the tissue of origin. Other carcinomas do not express some of the cytokeratins present in the tissue of their origin and, vice versa, certain components which are minor or apparently absent in normal tissue are major cytokeratins in the corresponding tumor. These differences may be explained by cell type selection during carcinogenesis, but changes of expression during tumor development cannot be categorically excluded. The possibility of cell type heterogeneity within a given tumor is also discussed. Similarly complex patterns of cytokeratin polypeptides have been noted in certain cultured human carcinoma cell lines (e.g., A-431, RPMI 2650, Detroit 562, A-549) and can also be observed in cel

Topics: Breast Neoplasms; Carcinoma, Squamous Cell; Cell Line; Cytoskeleton; Digestive System Neoplasms; Electrophoresis, Polyacrylamide Gel; Epithelium; Humans; Keratins; Liver Neoplasms; Lymphatic Metastasis; Neoplasms; Peptides; Rectal Neoplasms; Respiratory Tract Neoplasms; Urinary Bladder Neoplasms

In this study, we report the isolation of a Mr 52,000 keratin from a nontumorigenic bladder epithelial cell line and its localization, by immunofluorescence, in bladder frozen sections at different stages of neoplastic progression and in various carcinogen-transformed bladder epithelial cells and human bladder carcinoma-derived cell lines. The results showed that: (a) the Mr 52,000 keratin is present in basal epithelial cells of the normal bladder but absent from the intermediate and superficial epithelial cell layers; (b) hyperplastic bladder lesions, induced in mice after the administration of butyl-(4-hydroxybutyl)-nitrosamine, resulted primarily from the proliferation of cells in the basal compartment; (c) butyl-(4-hydroxybutyl)nitrosamine-induced bladder carcinomas displayed differential staining with the anti-Mr 52,000 keratin antiserum reflecting divergent differentiated status within a tumor and a subpopulation of cells with reduced overall keratin expression; (d) primary cultures of bladder carcinomas revealed a subpopulation of cells with limited filamentous keratin as was observed in vivo; (e) mouse bladder epithelial cell lines transformed in culture by a chemical carcinogen showed a loss or reduction in expression of the Mr 52,000 keratin; (f) two human bladder carcinoma cell lines displayed very limited expression of the Mr 52,000 keratin. Although the loss or reduction in expression of a specific keratin is unlikely to be responsible for transformation, it may contribute to the heterogeneity in the differentiated state and morphology of bladder epithelial cells during neoplastic progression both in vivo and in culture.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Cell Line; Cell Transformation, Neoplastic; Cricetinae; Epithelium; Fluorescent Antibody Technique; Humans; Immune Sera; Keratins; Mice; Microscopy, Phase-Contrast; Molecular Weight; Rats; Urinary Bladder; Urinary Bladder Neoplasms

A rabbit antiserum prepared against human keratins isolated from calluses was applied to sections of 108 neoplasms using indirect immunofluorescence and immunoperoxidase technics. The vast majority of epithelial neoplasms were strongly positive for keratin-type proteins, even in the absence of obvious keratinization or squamous differentiation as revealed by light microscopy. This keratin-positivity was invariably correlated with the identification of intermediate-sized filaments arranged in loose or dense bundles in the cytoplasm of neoplastic epithelial cells. Keratin-negative neoplasms included nevi, malignant melanomas, carcinoid tumors, malignant lymphomas, and a variety of connective-tissue tumors. Immunologic identification of keratin-type proteins was particularly helpful in establishing the epithelial nature of "undifferentiated" malignant tumors, including oat cell carcinomas.

Topics: Adenocarcinoma; Animals; Carcinoma; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Humans; Immunoenzyme Techniques; Keratins; Rabbits; Skin Neoplasms; Urinary Bladder Neoplasms

Immunoelectron microscope cytochemistry was carried out on 2% paraformaldehyde fixed, 50 mu sections of normal urothelium and bladder carcinoma cells in culture using antisera raised in rabbits to human 40-63 000 MW epidermal "broad spectrum" keratin and calf urothelial "luminal epithelial antigen" (aLEA) Both the unconjugated and indirect immunoperoxidase-DAB techniques were used before routine embedding. The localisation of both keratin and luminal epithelial antigen (LEA) was similar in normal and neoplastic cells and reaction product was associated not only with tonofilaments but also lining membrane vesicles and on fine filaments in the cytoplasmic ground substance.

Topics: Animals; Carcinoma; Epithelium; Humans; Keratins; Mice; Mice, Inbred C57BL; Microscopy, Electron; Neoplasms, Experimental; Rats; Rats, Inbred Strains; Urinary Bladder; Urinary Bladder Neoplasms

14 urinary bladder carcinomas of all main types were investigated with antisera to "broad spectrum keratin" (aK), "luminal epithelial antigen" (aLEA) and carcinoembryonic antigen (aCEA), using an indirect immunoperoxidase method on formalin fixed paraffin embedded sections. Keratin and LEA were both present in normal transitional epithelium, papilloma and carcinoma in situ whereas CEA was absent. Transitional cell carcinomas reacted with both aK and aLEA whereas CEA was seen only in a few foci. In squamous metaplasia and squamous carcinoma reaction with aK was particularly strong, while LEA was almost lacking and CEA was present in necrotic centres. In adenocarcinomas aK and aLEA reacted equally while aCEA reacted only on the surface.

Topics: Adenocarcinoma; Antigens, Neoplasm; Carcinoembryonic Antigen; Carcinoma in Situ; Carcinoma, Papillary; Carcinoma, Squamous Cell; Carcinoma, Transitional Cell; Epithelium; Humans; Immunoenzyme Techniques; Keratins; Urinary Bladder Neoplasms

The effect of 5-halogenated deoxyuridines on the keratinization of an in vitro-transformed rat bladder epithelial cell line, BES20B, was studied by a cover slip culture method established previously. 5-Bromodeoxyuridine (BrdU) and 5-iododeoxyuridine (IdU) both showed dose-dependent inhibition of the keratinization. The complete inhibition of keratinization was achieved by BrdU at concentrations higher than 5 microgram/ml. At these concentrations, BrdU reduced the saturation density of the test cells on a cover slip. Removal of BrdU from the culture medium restored the cell growth as well as the keratinization of BrdU-treated cells, suggesting that the effect of BrdU is reversible. The effect of BrdU was abolished by the addition of excess thymidine. A possible mechanism of the anti-keratinization effect of BrdU is discussed in comparison with that of vitamin A.

Topics: Animals; Binding, Competitive; Bromodeoxyuridine; Cell Line; Cell Transformation, Neoplastic; DNA Replication; Epithelium; Floxuridine; Idoxuridine; Keratins; Rats; RNA, Neoplasm; Thymidine; Urinary Bladder Neoplasms; Vitamin A

The terminal differentiation, keratinization, of a rat bladder tumor cell line, NBT II, occurred in multicellular aggregates. After aggregation, these cells did not undergo a round of mitosis before keratinization. 5-Bromodeoxyuridine added to the monolayer cell culture 2 days before aggregation completely prevented this differentiation; it was ineffective when added at the time of cell aggregation. Vitamin A prevented the keratinization of NBT II cells in aggregates but did not inhibit aggregate formation; it enhanced the number of cells engaged in DNA synthesis. This model appears to be very useful for analyzing the mechanisms of terminal differentiation and its modulation by vitamin A in tumor cells.

Topics: Animals; Bromodeoxyuridine; Carcinoma, Squamous Cell; Cell Aggregation; Cell Differentiation; Cell Line; Cytarabine; DNA, Neoplasm; Keratins; Mitosis; Rats; Urinary Bladder Neoplasms; Vitamin A

Cellular retinol-binding protein (CRBP) in cancers and in vitro-transformed epithelial cells of the ACI/N rat urinary bladder was assayed with radiolabeled retinol by sucrose density gradient centrifugation. CRBP was detected in abundance in tumor tissues of all 5 transplantation bladder cancer lines. However, this protein was below the limits of detection in bladder cancer cells of culture lines that had originated from the same primary cancers as the corresponding transplantation lines. CRBP was detected in small quantity in in vitro-transformed epithelial cells of a culture line but it was absent in another line of similar origin. The addition of retinol to the culture medium prevented the in vitro keratinization of CRBP-positive cells but it had no effect on the keratinization of CRBP-negative cells.

Topics: Animals; Cell Line; Cell Transformation, Neoplastic; Keratins; Neoplasm Transplantation; Neoplasms, Experimental; Rats; Retinol-Binding Proteins; Retinol-Binding Proteins, Cellular; Transplantation, Homologous; Urinary Bladder; Urinary Bladder Neoplasms

In vitro and in vivo transformed epithelial cells of the rat urinary bladder keratinized well on a coverslip culture. Keratinization proceeded more rapidly in in vitro-transformed cells than in in vivo-induced bladder cancer cells. The grade of keratinization was estimated from the absorption spectrum of a papanicolaou-stained specimen which afforded two peaks derived from keratinized cells and viable cells. Using this semiquantitative method, effect of various drugs on keratinization was studied on 3 lines of in vitro-transformed epithelial cells. Vitamin A and its analogs prevented the keratinization in 2 out of 3 lines at dosage over 1 micrograms/ml, but other drugs such as vitamin C, E, and K, steroid hormones, cyclic AMP, polyamines, polyanions, and dimethyl sulfoxide were ineffective for preventing keratinization. Amino acid compositions of keratinized cells and viable cells were not fundamentally different.

Topics: Amino Acids; Animals; Carcinoma, Squamous Cell; Cell Line; Cell Transformation, Neoplastic; Keratins; Male; Neoplasms, Experimental; Rats; Urinary Bladder Neoplasms; Vitamin A

A polypoid squamous cell carcinoma with pseudosarcomatous stroma of the urinary bladder was studied electron microscopically. The epithelial component was a typical squamous carcinoma that consisted of cells with abundant bundles of tonofilaments that converged toward well-developed desmosomes; keratohyalin granules were also seen. The stroma consisted of fusiform cells with dilated rough endoplasmic-reticulum cisternae and irregular cytoplasmic projections that were suggestive of active fibroblasts. No structures suggestive of an epithelial origin or of advanced mesenchymal differentiation were recognized. These observations are consistent with the notion that the pseudosarcomatous stroma represents a reactive process that is probably related to the growth of the epithelial neoplasm. Given the differences in behavior and prognosis between carcinomas with pseudosarcomatous stroma and true carcinosarcomas, efforts at separation of these entities are warranted.

Topics: Aged; Carcinoma, Squamous Cell; Cell Nucleus; Connective Tissue; Connective Tissue Cells; Cytoplasm; Endoplasmic Reticulum; Epithelial Cells; Epithelium; Humans; Keratins; Male; Mitochondria; Sarcoma; Urinary Bladder Neoplasms; Vacuoles

Topics: Adult; Aged; Carcinoma, Squamous Cell; Esophageal Neoplasms; Female; Granuloma; Humans; Keratins; Laryngeal Neoplasms; Male; Middle Aged; Mouth Neoplasms; Radium; Skin Diseases; Urinary Bladder Neoplasms; Uterine Cervical Neoplasms