lewis-x-antigen and Urinary-Bladder-Neoplasms

Bladder cancer continues to be one of the most common malignancies. Those who have been already diagnosed are at high risk for recurrence, especially if the pathology demonstrates high-grade disease. Diagnosis and surveillance is reliant on invasive evaluation with cystoscopy. Urinary cytology has been used to aid in diagnosis, but its use is limited. Other assays have been developed that may aid in clinical decision making. The ultimate goal will be the development of a highly sensitive and specific urinary marker for bladder cancer. This would provide a noninvasive means of diagnosing the disease and limit the number of unnecessary cystoscopies. This article will review the currently available urinary bladder cancer markers. It will also review new and investigational urinary markers that have shown promise for future clinical use.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Humans; Hyaluronic Acid; In Situ Hybridization, Fluorescence; Lewis X Antigen; Microsatellite Repeats; Nuclear Proteins; Sensitivity and Specificity; Telomerase; Urinary Bladder Neoplasms; Urine

Bladder cancer has a very high frequency of recurrence and therefore requires lifelong surveillance, traditionally consisting of serial cystoscopy and cytology. These tests are both invasive and expensive, with considerable inter-user and inter-institutional variability. In addition, the sensitivity of cytology in detecting low-grade tumors is low. Therefore, there has been active investigation into urinary biomarkers that can either supplement or supplant these tests. At this point there are only six urine-based tests that are FDA-approved in bladder cancer surveillance, but a wide variety of other biomarkers are being studied. In this review, we examine the natural history of bladder cancer as well as the rationale and performance of an ideal urinary biomarker. The authors describe the FDA-approved biomarkers such as Bladder Tumor Antigen, ImmunoCyt, Nuclear Matrix Protein-22, and Fluorescent In Situ Hybridization, as well as the most promising investigational tests (i.e., Urinary bladder cancer test, BLCA-1, BLCA-4, hyaluronic acid, hyaluronidase, Lewis X antigen, microsatellite analysis, Quanticyt, soluble Fas, Survivin, and telomerase). The biological foundation, methodologies, and diagnostic performance of the biomarkers are discussed. The characteristics of the biomarkers are compared to urine cytology. At this time, urine biomarkers are utilized in a variety of clinical situations but their role is not well defined. The goal of identifying an optimal marker that will replace cystoscopy and/or cytology is still ongoing.

Topics: Adjuvants, Immunologic; Biomarkers, Tumor; Carcinoma, Transitional Cell; Cysteine Proteinase Inhibitors; Cystoscopy; Fas Ligand Protein; Humans; Hyaluronic Acid; Hyaluronoglucosaminidase; In Situ Hybridization, Fluorescence; Inhibitor of Apoptosis Proteins; Lewis X Antigen; Microtubule-Associated Proteins; Nuclear Proteins; Population Surveillance; Prognosis; Sensitivity and Specificity; Survivin; Telomerase; Urinary Bladder Neoplasms

A study was made to determine the sensitivity and specificity of immunostaining of the Lewis X antigen in exfoliated urothelial cells from voided urine, for the detection and surveillance of bladder tumors.. Three consecutive voided urine specimens were obtained from 101 patients, 78 of whom were under surveillance because of a history of bladder tumors, and 23 were being evaluated because of hematuria or irritative urinary symptoms. Indirect immunoperoxidase staining of two urine samples was done on cytocentrifuge slides, using the P12 monoclonal antibody against the Lewis X antigen. The diagnosis of the presence of urothelial tumor was made if more than 5% of the cells showed a typical red-brown staining. Cytopathologic examination of the third urine specimen was done according to Papanicolaou. Each patient underwent cystoscopy, and biopsies were obtained whenever there was endoscopic evidence of bladder tumors or carcinoma in situ.. Cystoscopy and biopsies revealed transitional cell carcinoma in 32 patients, whereas 69 patients had no evidence of bladder tumors. Immunocytology of one urine sample showed true-positive results in 26 of the 32 patients with bladder tumors, corresponding to a sensitivity of 81.25%. When two samples were examined, a sensitivity of 97% and a specificity of 85.5% were obtained. When the results of cytology and immunocytology were combined, sensitivity reached 100%. High-grade and low-grade transitional cell tumors were detected with equal efficiency.. The use of P12 monoclonal antibody for evaluation of Lewis X reactivity in cytologic preparations from multiple voided urine specimens can improve the sensitivity of noninvasive detection of bladder cancer. The technique may ultimately replace cystoscopy in monitoring therapeutic response and tumor recurrence.

Topics: Biomarkers, Tumor; Carcinoma in Situ; Carcinoma, Transitional Cell; Humans; Immunoenzyme Techniques; Lewis X Antigen; Predictive Value of Tests; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urine

MicroRNAs (miRNAs) have been widely studied in various human cancers, including bladder cancer. Previous report revealed that miR-125a-5p is downregulated in urothelial carcinomas. However, the biological function and molecular mechanism of miR-125a-5p in bladder cancer has not been elucidated. Therefore, this study focused on the role of miR-125a-5p in bladder cancer. The expression levels of miR-125a-5p were firstly tested in one normal cell line and four bladder cancer cell lines with qRT-PCR. The relative lower expression of miR-125a-5p was detected in bladder cancer cells. To confirm the effects of ectopic expression of miR-125a-5p on the biological behaviors of bladder cancer cells, gain-of-function assays were carried out. According to experimental results, miR-125a-5p overexpression suppressed cell proliferation and cell cycle progression, induced cell apoptosis. Moreover, overexpression of miR-125a-5p suppressed cell migration and invasion and reversed epithelial-mesenchymal transition (EMT). Mechanism investigation indicated that FUT4 is a target mRNA of miR-125a-5p in bladder cancer. The effects of FUT4 on cell proliferation, apoptosis, migration and invasion were identified by conducting gain-of-function assays. Finally, rescue assays indicated that FUT4 can reverse the effects of miR-125a-5p on bladder cancer progression. In summary, miR-125a-5p suppresses bladder cancer progression through targeting FUT4.

Topics: Apoptosis; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Down-Regulation; Epithelial-Mesenchymal Transition; Fucosyltransferases; Gene Expression Regulation, Neoplastic; Humans; Lewis X Antigen; MicroRNAs; RNA, Messenger; Urinary Bladder Neoplasms

Preclinical studies suggest that signal transducer and activator of transcription 3 (STAT3)-mediated recruitment of neutrophils to premetastatic tissue occurs prior to metastatic progression.. We sought to determine if neutrophilic infiltration in benign nodal tissue is associated with poor clinical outcome in patients with muscle-invasive bladder cancer.. Formalin-fixed, paraffin-embedded tissue was secured from 55 patients with muscle-invasive bladder cancer who had undergone cystectomy at our institution. Sections of benign lymph nodes were obtained and stained with primary antibodies against 3-fucosyl-N-acetyl-lactosamine, phosphorylated STAT3, and interleukin-17, the latter being a key mediator of neutrophil infiltration and STAT3 activation.. The Kaplan-Meier method was used to interrogate differences in overall survival (OS) in patients with high versus low biomarker expression. Cohorts stratified by receipt and nonreceipt of neoadjuvant chemotherapy were separately explored.. Of the 55 patients examined, 19 patients (35%) had no prior neoadjuvant chemotherapy. Amongst these patients, median OS was improved in patients with low 3-fucosyl-N-acetyl-lactosamine. The results herein support the hypothesis that bladder cancer metastasis may be driven by STAT3-mediated neutrophilic infiltration in premetastatic sites. Validation of these findings using tissues derived from a phase 3 surgical trial (Southwest Oncology Group 1011) is currently underway.. Lymph node metastases occur in up to 25% of patients with muscle-invasive cancer and it represents one of the most frequent sites of bladder cancer metastasis. This report provides preliminary evidence that neutrophil levels in benign lymph nodes may predict clinical outcome.

Topics: Adult; Aged; Aged, 80 and over; Chemotherapy, Adjuvant; Female; Humans; Immunohistochemistry; Interleukin-17; Kaplan-Meier Estimate; Leukocyte Count; Lewis X Antigen; Lymph Nodes; Male; Middle Aged; Neoadjuvant Therapy; Neoplasm Invasiveness; Neutrophils; Phosphorylation; Prognosis; STAT3 Transcription Factor; Survival Rate; Urinary Bladder; Urinary Bladder Neoplasms

It is widely accepted that sialyl Lewis X (sLeX) and sialyl Lewis A (sLeA, also known as CA 19-9) glycans expressed on cancer cells function in E-selectin-mediated metastasis. Recently, it was reported that 6-sulfo sLeX glycans detected by the MECA-79 monoclonal antibody are expressed in roughly a quarter of gastric adenocarcinoma cases, and that these cases show a poorer prognosis than MECA-79-negative cases do. The present study was undertaken to assess expression of 6-sulfo sLeX glycans in bladder urothelial carcinoma and evaluate potential clinical implications.. We analyzed 78 specimens representing bladder urothelial carcinoma, as well as 4 bladder urothelial carcinoma cell lines, by immunostaining with a battery of anticarbohydrate antibodies. We also undertook an E-selectin·IgM chimera binding assay to assess E-selectin binding to 6-sulfo sLeX expressed on bladder urothelial carcinoma cells and performed reverse transcription polymerase chain reaction and complementary DNA transfection to determine which N-acetylglucosamine-6-O-sulfotransferases function in 6-sulfo sLeX biosynthesis in those cells. Finally, we performed double-immunofluorescence staining for MECA-79 and either CD3 or CD8 to evaluate potential association between high endothelial venule (HEV)-like vessels and tumor-infiltrating T lymphocytes.. 6-Sulfo sLeX glycans were expressed in ~20% of bladder urothelial carcinoma cases, particularly in plasmacytoid and micropapillary variants. Positive cells were also bound by E-selectin·IgM chimeras in a calcium-dependent manner. Transcripts encoding N-acetylglucosamine-6-O-sulfotransferase-2 were detected preferentially in HT-1197 bladder urothelial carcinoma cells expressing 6-sulfo sLeX, and transfection of the enzyme complementary DNA into HT-1376 cells, which do not express 6-sulfo sLeX glycans, resulted in cell surface expression of 6-sulfo sLeX. Furthermore, 6-sulfo sLeX glycans were expressed in HEV-like vessels induced in and around lymphocyte aggregates formed near carcinoma cell nests. These HEV-like vessel-associated tumor-infiltrating lymphocytes were composed primarily of CD3(+) T cells, with a fraction of CD8(+) cytotoxic T cells.. Our findings indicate that 6-sulfo sLeX glycans likely play 2 roles in bladder urothelial carcinoma progression: one in lymphocyte recruitment to enhance antitumor immune responses, and the other in E-selectin-mediated tumor cell adhesion to vascular endothelial cells, which is potentially associated with metastasis.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Carbohydrate Sulfotransferases; E-Selectin; Female; Fluorescent Antibody Technique; Follow-Up Studies; Humans; Immunoenzyme Techniques; Lewis X Antigen; Male; Middle Aged; Neoplasm Staging; Oligosaccharides; Prognosis; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sialyl Lewis X Antigen; Sulfotransferases; Urinary Bladder Neoplasms

Both cancer-related inflammation and tumor-induced immune suppression are associated with expansion of myeloid cell subsets including myeloid-derived suppressor cells. However, little known regarding characteristics of myeloid cells in patients with bladder cancer. In this study, we analyzed myeloid cells from peripheral blood (PBMC) and tumor tissue that were collected from patients with superficial noninvasive and invasive urothelial carcinomas. Our results demonstrate that PBMC from bladder cancer patients contain two major CD11b myeloid cell subsets: granulocyte-type CD15(high) CD33(low) cells and monocyte-type CD15(low) CD33(high) cells. The number of circulating granulocytic but not monocytic myeloid cells in cancer patients was markedly increased when compared to healthy individuals. Both myeloid cell subsets from cancer patients were highly activated and produced substantial amounts of proinflammatory chemokines/cytokines including CCL2, CCL3, CCL4, G-CSF, IL-8 and IL-6. Granulocytic myeloid cells were able to inhibit in vitro T cell proliferation through induction of CD4(+) Foxp3(+) T regulatory cells. Analysis of bladder cancer tissues revealed that tumors were infiltrated with monocyte-macrophage CD11b(+) HLA-DR(+) and granulocytic CD11b(+) CD15(+) HLA-DR(-) myeloid cells. Collectively, this study identifies myeloid cell subsets in patients with bladder cancer. We demonstrate that these highly activated inflammatory myeloid cells represent a source of multiple chemokines/cytokines and may contribute to inflammation and immune dysfunction in bladder cancer.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; CD11 Antigens; Cytokines; Granulocytes; Humans; Immune Tolerance; Lewis X Antigen; Lymphocyte Activation; Monocytes; Myeloid Cells; Sialic Acid Binding Ig-like Lectin 3; Urinary Bladder Neoplasms

Carbohydrate antigens and E-selectin play important roles in the invasion and metastasis of cancers. We examined the expression of these antigens and their ligand protein, E-selectin, in urothelial carcinomas to evaluate whether their staining is correlated with the grade and stage of cancer. We studied the expression of carbohydrate antigens (type 1 and type 2 blood-group antigens) and E-selectin in urothelial carcinomas of the renal pelvis, ureter, and urinary bladder in 52 patients by staining SSEA-1 (LeX), sialyl LeX (sLeX), DU-PAN-2, CA19-9, and E-selectin with 5 different monoclonal antibodies (MAbs) to evaluate whether their staining correlated with cancer grade and stage. The differences between organs with regard to the degree of expression of these antigens were not evident. Type 2 antigens (SSEA-1 and sialyl LeX) are frequently expressed in the tumor cells regardless of atypical grade. The expression level of type 1 antigens (DU-PAN-2 and CA19-9) is lower than that of type 2 antigens. However, the presence of DU-PAN-2 tends to correlate with the grade of atypia; however, that of CA19-9 is inversely proportional to the grade of atypia. The lack of CA19-9 and appearance of DU-PAN-2 in urothelial carcinoma implies a high malignant potential. The expression of E-selectin can be correlated with stage and grade of tumor atypia. Type 2 antigen and E-selectin may be involved in tumor invasion and metastasis.

Topics: Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; CA-19-9 Antigen; E-Selectin; Female; Humans; Immunohistochemistry; Kidney Neoplasms; Lewis X Antigen; Male; Middle Aged; Oligosaccharides; Sialyl Lewis X Antigen; Ureteral Neoplasms; Urinary Bladder Neoplasms; Urologic Neoplasms; Urothelium

A problem in the interpretation of noninvasive urine tests for detection of bladder carcinoma is the finding of false-positive results. Several authors have described that patients with false-positive results are at high risk for tumor recurrence or progression. Only few data are available for comparing the clinical course of patients with false-positive test results and patients with true-negative results. We studied whether patients with false-positive results of various urine test had a higher recurrence rate than patients with true-negative results.. Urine samples from 61 patients without evidence of active bladder carcinoma were included. Of the 61 patients, 51 had a history of bladder cancer, and 10 underwent transurethral resection for suspect of bladder carcinoma but had negative pathologic findings. Immunocytology (Lewis X and 486p3/12) was performed on bladder washings, and BTAstat and NMP22 were performed on urine samples.. During the follow-up period, 22 patients had one or more false-positive BTAstat test results, 25 patients had one or more false-positive NMP22 tests, 42 patients had at least one false-positive Lewis X test, and 11 patients had one or more false-positive 486p3/12 test. During a follow-up period of 3-39 months (median, 17.6 months) four patients expected a tumor recurrence. Among patients with false-positive urine test results 2 of 22 (9.1%, BTAstat), 2 of 25 (8%, NMP22), 4 of 42 (9.5%, Lewis X), and 3 of 11 (27.2%, 486p3/12) suffered from tumor recurrence. In contrast, among patients with true-negative test results 2 of 39 (5.2%, BTAstat), 2 of 36 (5.6%, NMP22), 0 of 18 (0%, Lewis X), 1 of 50 (2.0%, 486p3/12) had a tumor recurrence.. Patients with a false-positive urine test result do not generally have a greater risk of tumor recurrence or progression than patients with a true-negative result. In our series, only patients with false-positive 486p3/12 test result had a higher recurrence rate. Our findings do not justify a more aggressive adjuvant treatment or surveillance for patients with false-positive urine tests.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; False Positive Reactions; Follow-Up Studies; Humans; Immunoenzyme Techniques; Lewis X Antigen; Neoplasm Staging; Nuclear Proteins; Predictive Value of Tests; Prognosis; Urinary Bladder Neoplasms

The non-invasive detection of urothelial carcinoma remains challenging. The aim of this study was the prospective evaluation of urine markers for bladder carcinoma. We compared the NMP 22 and BTAstat tests with immunocytology (IC) using monoclonal antibodies against the Lewis X antigen and against 486p3/12.. NMP 22 and BTAstat were performed on urine samples, and IC with 486p3/12 and Lewis X staining was performed on urine samples as well as bladder wash specimens ( n=146) in patients ( n=115) undergoing transurethral resection on suspicion of bladder cancer (70 specimens) or follow up cystoscopy because of a history of bladder cancer (76 specimens). Bladder cancer was detected in 54 patients (pTa: n=25, pT1: n=20, pT2: n=8, CiS: n=1). Cut-off levels were 10 U/ml for the NMP 22, 30% positive cells for 486p3/12, and 5% positive cells for the Lewis X test.. The BTAstat test was positive in 65 (44.5%) cases, the NMP 22 in 69 (47.3%) cases, IC with 486p3/12 and the Lewis X was positive in 52 (35.6%) and 109 (74.7%) cases, respectively. Sensitivity was 70.3% (BTAstat), 68.5% (NMP 22), 94.4% (Lewis X), and 68.5% (486p3/12), respectively. The specificity was 70.6% (BTAstat), 65.2% (NMP 22), 36.9% (Lewis X), and 83.6% (486p3/12), respectively. Among the patients with a false positive test 2/22 (9.0%) patients (BTAstat), 2/25 (8%) patients (NMP 22 test), 4/43 (9.3%) patients (Lewis X), and 3/11 (27%) patients (486p3/12), respectively, suffered from tumor recurrence. In contrast, among the patients with a correct negative test 2/39 (2.0%) (BTAstat), 2/36 (0.5%) (NMP 22), 0/18 (0%) (Lewis X), and 1/50 (2.0%) (486p3/12), respectively, suffered from tumor recurrence.. IC with the Lewis X revealed a higher sensitivity than all of the tested, commercially available methods. Because of its high sensitivity and its high negative predictive value, the Lewis X test may be useful for screening a high-risk population. Patients with a false positive 486p3/12 test have an increased risk of tumor recurrence when compared with patients with a correct negative test.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; Biopsy; Carcinoma, Transitional Cell; Cystoscopy; Humans; Lewis X Antigen; Neoplasm Recurrence, Local; Neoplasm Staging; Nuclear Proteins; Predictive Value of Tests; ROC Curve; Urinary Bladder; Urinary Bladder Neoplasms; Urine

The noninvasive detection of urothelial carcinoma remains challenging. We prospectively evaluated urine markers for bladder carcinoma. We compared the NMP22 (Matritech, Cambridge, Massachusetts) and BTA Stat (Bard Diagnostics, Redmond, Washington) tests with immunocytology using mAbs 486p3/12 and BG7 against Lewis X antigen.. The NMP22 and BTA Stat tests were performed in urine samples and immunocytology with mAbs 486p3/12 and BG7 staining were performed in bladder washing specimens in 146 samples of 115 patients undergoing transurethral resection for suspected bladder carcinoma (70) or 45 undergoing followup cystoscopy for a history of bladder carcinoma (76). Bladder carcinoma was detected in 54 patients, including stages pTa in 25, pT1 in 20, pT2 in 8 and carcinoma in situ in 1, while 61 had no evidence of bladder carcinoma. The cutoff was 10 units per ml. for the NMP22, 30% positive cells for 486p3/12 and 5% positive cells for the Lewis X tests.. BTA Stat was positive in 65 samples (44.5%) and NMP22 was positive in 69 (47.3%). Immunocytology with mAbs 486p3/12 and BG7 against Lewis X was positive in 52 (35.6%) and 109 (74.7%) samples, respectively. Sensitivity was 70.3% for BTA Stat, 68.5% for NMP22, 68.5% for 486p3/12 and 94.4% for Lewis X. Specificity was 70.6% for BTA Stat, 65.2% for NMP22, 83.6% for 486p3/12 and 36.9% for Lewis X. Area under the receiver operating characteristics curve was 0.6804 for NMP22, 0.7226 for Lewis X and 0.8002 for 486p3/12. False-positive results on BTA Stat in 2 of 22 patients (9%), on NMP22 in 2 of 25 (8%), on 486p3/12 in 3 of 11 (27%) and on Lewis X in 4 of 43 (9.3%) were associated with tumor recurrence. Furthermore, negative results on BTA Stat in 2 of 39 patients (2%), on NMP22 in 2 of 36 (0.5%), on Lewis X in 0 of 18 (0%) and on 486p3/12 in 1 of 50 (2%) was associated with tumor recurrence during followup.. Immunocytology with mAbs against Lewis X showed higher sensitivity than all commercially available tests evaluated. Because of its high sensitivity and high negative predictive value, it may be useful for screening in a high risk population. Patients with a false-positive 486p3/12 test results are at increased risk for tumor recurrence compared with those with negative results.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Biomarkers, Tumor; Biopsy; Carcinoma, Transitional Cell; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Lewis X Antigen; Male; Neoplasm Staging; Nuclear Proteins; Predictive Value of Tests; Urinary Bladder; Urinary Bladder Neoplasms

DNA-image-cytometry and antibodies directed against the Lewis X- and the 486p 3/12 antigen were applied to improve diagnostic accuracy of urinary cytology for the detection of bladder cancer. Cytology, immunocytology and DNA-image-cytometry were performed in spontaneously voided urine samples and barbotage bladder washings from 71 patients. The DNA content was determined using the CM-1 Cytometer according to the recommendation of the ESCAP Consensus Report on Standardization of DNA-image-cytometry (1995). For immunocytological examination we used the monoclonal anti Lewis X antibody P-12 and antibody 486p 3/12. All patients underwent subsequent cystoscopy and for any suspicious lesion biopsy or transurethral resection was done. Histological findings revealed 31 patients with transitional cell carcinomas of different stages and grades of malignancy. 40 patients had various benign diseases of the urinary bladder. Cytology yielded a sensitivity of 68% and a specificity of 100%. DNA aneuploidy was detected in 81% of cancer patients with a specificity of 100%. By combination of these two methods the overall sensitivity increased to 87%. Immunocytology with Lewis X and 486p 3/12 antibodies showed reactivity in 84% and 87% in combination with a specificity of 80% and 70%, respectively. By combining urinary cytology, immunocytology and/or DNA-image-cytometry the overall sensitivity increased to 94% with no change in specificity. DNA-image-cytometry should be used to evaluate particularly urothelial cells suspicious for malignancy in urinary specimens. Because of low specificity the monoclonal antibodies against Lewis X- and 486p 3/12 antigens are not helpful in screening for bladder cancer. Nevertheless, their high sensitivity may justify their use in case DNA image cytometry is not available and in the follow up of patients with transitional cell carcinoma.

Topics: Aged; Carcinoma, Transitional Cell; Female; Humans; Image Cytometry; Immunohistochemistry; Lewis X Antigen; Middle Aged; Observer Variation; Reproducibility of Results; Sensitivity and Specificity; Urinary Bladder Neoplasms; Urine; Vaginal Smears

To assess the contribution of the carbohydrate antigens, sialyl-Lewis X (sLe(x)) and sialyl-Lewis A (sLe(a)), which are known to be ligands for E-selectin, to the adhesion between human urothelial cancer cells and cytokine-activated human endothelial cells.. We studied the expression of sLe(x) and sLe(a) antigens of three bladder cancer cell lines (JTC 30, JTC 32, and T24) by flow cytometry and the adherence to interleukin 1beta-activated human umbilical vein endothelial cells (HUVEC).. JTC 30 and JTC 32 cells expressed both sLe(x) and sLe(a) antigens, and showed adhesion to activated HUVEC, which was completely abolished by anti-E-selectin antibody. T24 cells expressed neither sLe(x) nor sLe(a) antigen, and did not adhere to activated HUVEC. Each of anti-sLe(a) or anti-sLe(x) antibody partially blocked the attachment of JTC 30 cells to activated HUVEC, and combination of these antibodies almost completely blocked the adhesion. The combination of antibodies did not significantly influence the adhesion of JTC 32 cells.. These results indicate that both sLe(a) and sLe(x) carbohydrate antigens are involved in E-selectin-mediated adhesion of some urothelial cancers, and that there might be unknown ligands for E-selectin on urothelial cancer cells.

Topics: Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Cell Adhesion; Cell Adhesion Molecules; Endothelium, Vascular; Gangliosides; Humans; Interleukin-1; Lewis X Antigen; Oligosaccharides; Sialyl Lewis X Antigen; Tumor Cells, Cultured; 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

To improve sensitivity and specificity of urinary cytology for bladder cancer cell detection we used deoxyribonucleic acid (DNA) image cytometry and the monoclonal anti-Lewis X antibody P12.. Spontaneously voided urine and additional barbotage bladder washings of 25 patients with transitional cell carcinomas and 25 patients with benign diseases of the bladder were analyzed by conventional cytology, immunocytology and DNA image cytometry. The DNA content was determined in specimens stained with Feulgen according to the European Society for Analytical Cellular Pathology consensus report on standardization of DNA image cytometry. For the immunocytological examination we used the avidin-biotin-complex immunoperoxidase method with the monoclonal antibody P12 directed against the Lewis X determinant.. The cytological examination revealed a sensitivity of 72% and a specificity of 100%. By using DNA image cytometry with Kolmogoroff-Smirnow test sensitivity increased up to 84% with a specificity of 100%. The immunocytological examination with Lewis X showed a sensitivity of 84% and a specificity of 80%.. Combining cytology and DNA cytometry the overall sensitivity increased to 92% and with the additional application of immunocytology for Lewis X antigen sensitivity it increased to 96% but was accompanied by a decrease in specificity to 80%. DNA image cytometry and Lewis X antigen detection are not suitable for screening but suspicious urothelial cells in urinary cytology specimens can be evaluated specifically by DNA measurements. In the future it needs to be clarified whether DNA image cytometry in combination with Lewis X antigen detection can help to signal relapse and progression of transitional cell carcinoma of the bladder.

Topics: Aged; Carcinoma, Transitional Cell; DNA; Humans; Image Cytometry; Lewis X Antigen; Middle Aged; Sensitivity and Specificity; Urinary Bladder Neoplasms

We examined the use of immunostaining of the Lewis X antigen in exfoliated cells from voided urine samples, cytopathology and bladder ultrasound for noninvasive detection of bladder tumors as a potential substitute for cystoscopy.. A total of 260 patients were included, of whom 80 were evaluated because of irritative symptoms or hematuria and 180 were examined during followup visits after resection of bladder tumors. Voided urine samples were obtained from each patient for immunocytology and cytopathology. Bladder ultrasound and cystoscopy were performed. Biopsies were obtained whenever a bladder tumor was seen or if carcinoma in situ was suspected. Indirect immunoperoxidase staining was done on cytocentrifuge slides, using the P12 monoclonal antibody against the Lewis X antigen.. Cystoscopy and biopsies revealed bladder tumors in 84 patients. Immunocytology of 1 urine sample resulted in a sensitivity of 79.8% and a specificity of 86.4%. The diagnosis of primary carcinoma in situ by immunocytology was correct in 100% of the cases. The examination of 2 consecutive urine samples detected 95.1% of the tumors. False-negative results occurred in a few cases with small, superficial, low grade tumors. Cytopathology and bladder ultrasound resulted in a sensitivity of 47.6 and 66.7%, and a specificity of 97.7 and 97.2%, respectively. The results of immunocytology of 2 urine samples were equivalent to the combination of immunocytology of a single urine sample, cytology and ultrasound.. Immunostaining of the Lewis X antigen is significantly more sensitive than cytopathology for the detection of low grade as well as high grade tumor cells in voided urine. Immunocytological evaluation of 2 consecutive voided urine specimens for the Lewis X antigen is the most sensitive method currently available for noninvasive detection of transitional cell tumors. This assay may replace cystoscopy for detection of bladder cancer.

Topics: Humans; Immunohistochemistry; Lewis X Antigen; Sensitivity and Specificity; Ultrasonography; Urinary Bladder Neoplasms; Urine

Topics: Cystoscopy; Humans; Lewis X Antigen; Urinary Bladder Neoplasms

The expression of stage-specific embryonic antigen 1 (SSEA-1) in transitional cell carcinomas of the bladder (TCCB) has been reported to correlate with tumor grade and the likelihood of lymphatic metastases. We examined the expression of this antigen in TCCBs to evaluate if staining correlated with grade, stage, recurrence, progression and response to intravesical chemotherapy. We studied the expression of SSEA-1 in TCCBs from 74 patients by staining with two different monoclonal antibodies (Mabs), P-12 and anti-SSEA-1, to evaluate if staining correlated with grade, stage and tumor recurrence. Staining was considered as positive or negative irrespective of the intensity of staining. Extent of tumor staining was measured in quartiles of 100% (25, 50, 75 and 100). Follow-up was available in 47/74 (63%) patients and ranged from 6 months to 13 years (median 2 years). Staining with one or both Mabs was observed in 57/75 (76%) tumors. None of the grade I tumors showed > 50% staining while 26% of grade II and only 33% of grade III tumors showed staining of > 50% of cells. 21% of patients whose tumors showed staining with both Mabs were free of recurrence after resection of the primary tumor; 41% of patients with tumors staining negative with both Mabs showed no recurrence. Expression of SSEA-1 does correlate with tumor recurrence especially in grade II tumors, but the correlation is not very strong (0.05 < p < 0.1). Expression of this antigen is a weak indicator of recurrence in superficial TCCBs.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Transitional Cell; Humans; Immunohistochemistry; Kidney; Lewis X Antigen; Neoplasm Recurrence, Local; Retrospective Studies; Urinary Bladder; Urinary Bladder Neoplasms

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

Immunodetection of the Lewis X antigen has been suggested as a useful adjunct in the diagnosis of transitional cell carcinoma. To determine the specificity of Lewis X antigen immunostaining in this setting, we studied routinely processed, formalin-fixed tissue from 38 transitional cell carcinomas and 42 nonneoplastic urothelial lesions using the P12 monoclonal antibody to Lewis X antigen and an avidin-biotin immunohistochemical technique. Because Lewis X immunostaining of occasional umbrella cells has previously been noted in normal urothelium, staining of umbrella cells was not considered sufficient for a positive reaction in our study. Immunoreactivity for Lewis X antigen was seen in 34 of 38 (89%) cases of transitional cell carcinoma, in three of three cases of urothelial dysplasia, and in 20 of 39 (51%) of the reactive urothelial lesions. We conclude that immunostaining for the Lewis X antigen is not specific in the diagnosis of transitional cell carcinoma.

Topics: Antibodies, Monoclonal; Antibody Specificity; Avidin; Biomarkers, Tumor; Biotin; Carcinoma, Transitional Cell; Diagnosis, Differential; Epithelium; Humans; Immunohistochemistry; Lewis X Antigen; Retrospective Studies; Urinary Bladder; Urinary Bladder Neoplasms

Expression of binding sites for fucose binding proteins (FBP) of Lotus tetragonolobus were immunohistochemically analyzed in surgically extirpated specimens from patients with transitional cell carcinoma of the bladder. The degree of expression of FBP binding sites in the primary cancerous region correlated with the occurrence of lymph node metastasis. Thus, lymph node metastases occurred in 15 of 34 cases with high expression of FBP binding sites, but did not in 17 cases with low or no FBP binding site expression. All metastatic lymph nodes strongly reacted with FBP. In addition, primary lesions at the pT3b or pT4 stage more frequently reacted with FBP as compared with those at the pTa, pT1 or pT2 stage. Although FBP is known to react both with Gal beta 1----4(Fuc alpha 1----3)GlcNAc and Fuc alpha 1----2Gal sequences, comparative staining of other carbohydrate markers revealed that the latter structure is not related with metastatic potential and stages.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Transitional Cell; Female; Fucose; Humans; Lectins; Lewis X Antigen; Male; Middle Aged; Neoplasm Staging; Phenotype; Plant Lectins; Protein Binding; Urinary Bladder Neoplasms

Expression of stage-specific embryonic antigen-1 (SSEA-1) was immunohistochemically analyzed in primary cancerous regions from transitional cell carcinoma of the bladder. Lymph node metastasis occurred in 14 (50%) of 28 patients with high expression of SSEA-1 antigen, but in only one (4%) of 23 patients with low or no expression. Of 36 invasive bladder carcinoma patients with no metastasis at operation, 8 (57%) of 14 with high expression of SSEA-1 antigen died of cancer or had metastasis, whereas 4 (18%) of 22 with low or no expression had progression of the disease. In addition, stage pT3b-4 tumors more frequently reacted with SSEA-1 MAb than stage pT1 tumors (p less than 0.01). Grade 2 or 3 tumors were also more reactive than grade 1 tumors, which were all unreactive except for one (p less than 0.01, respectively). Our data suggest that SSEA-1, which is a Le(x) antigen, correlates with stage and grade as well as metastatic phenotype of transitional cell carcinoma of the bladder.

Topics: Carcinoma, Transitional Cell; Humans; Lewis X Antigen; Lymphatic Metastasis; Neoplasm Staging; Urinary Bladder Neoplasms

Sialyl dimeric Le(x) antigen was expressed in significant portion of transitional cell carcinoma of the human urinary bladder, but not in the normal uroepithelial tissue. Primary tumors with weak or no expression of the antigen scarcely metastasized to lymph nodes, whereas tumors with high levels of antigen expression metastasized frequently. Metastatic lymph nodes expressed the antigen in most cases. Sialyl dimeric Le(x) antigen was mainly located on 60 and 42 KDa glycoproteins. Since a group of cell adhesion molecules, called LECCAMs, recognize a portion of the antigen, the above results strongly suggest that a LECCAM on the surface of host cells recognizes the carbohydrate structure on the glycoprotein, leading to promotion of metastasis.

Topics: Biomarkers, Tumor; Carcinoma, Transitional Cell; Cell Membrane; Glycoproteins; Humans; Lewis X Antigen; Lymph Nodes; Lymphatic Metastasis; Molecular Weight; Neoplasm Metastasis; Urinary Bladder Neoplasms

The biopsy specimens of 91 patients between the ages of 38 and 94 with transitional cell carcinoma of the bladder were retrospectively reviewed to determine if the expression of CD15 antigen detected by a monoclonal antibody MC2 was correlated with prognosis. Expression was variable, ranging from strong expression of the antigen by only the superficial cells in well differentiated papillary lesions to weak expression by most cells in solid or invasive tumours. In the invasive component there was a correlation between MC2 expression and tumour type, suggesting that the cell surface carbohydrate detected by MC2 may have a role in cell adhesion. There was no correlation between staining and survival. It is concluded that tumour type, grade, and stage remain the best prognostic indicators of urothelial tumours.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation; Antigens, Differentiation, Myelomonocytic; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Transitional Cell; Humans; Lewis X Antigen; Middle Aged; Prognosis; Urinary Bladder Neoplasms