Progression Of Transitional Cell Carcinoma Research Articles

The relationship between ABO blood group type and tumor grade, stage, recurrence and progression in transitional cell carcinoma of the bladder: A preliminary report

Objectives: ABO blood grouping is a well-proven prognostic factor in many malignancies. This study aims to study the association and impact of ABO blood group on disease recurrence and progression in bladder carcinoma. Material and methods: Patients with bladder carcinoma undergoing transurethral resection of bladder tumor (TURBT) were studied prospectively for at least 1-year follow-up for recurrence and progression of the disease. Demographic profile along with blood group was noted. Results: Two hundred patients were included in the study and 194 patients were included in the final analysis. Muscle-invasive bladder cancer was present in 39 (20.1%) patients and the high-grade tumor was present in 88 (45.3%) patients. There was no statistical significance between the association of blood grouping and grade ( p=0.29) and stage of the disease ( p=0.20). During the follow-up period, there were 100 (64.5%) recurrences and 15 (9.7%) patients with non-muscle-invasive bladder carcinoma had progression. The association of blood group with recurrence ( p=0.66) and progression ( p=0.11) of disease was not statistically significant. Conclusion: There is no association between bladder cancer and ABO blood group in terms of grade and stage of the disease. The recurrence and progression of the disease did not differ significantly in different blood groups. Level of Evidence: 2b

The Prognostic Value of some Epithelial-Mesenchymal Transition Markers and Metastasis-Related Markers in Human Transitional Cell Carcinoma of the Bladder

Recent reports provided evidence that epithelial to mesenchymal transition (EMT) and some matrix metalloproteinases (MMPs) contribute to the invasion and metastasis of cancer cells. This study investigated the expression pattern of some EMT markers (E-cadherin and Vimentin) and some MMPs (MMP-2 and MMP-9) in transitional cell carcinoma (TCC). Fifty five paraffin embedded biopsies were included in this study. Expression pattern of E-cadherin and Vimentin was evaluated by immunohistochemistry while cytoplasmic mRNA expression of both MMP-2 and MMP-9 were determined by in situ hybridization. The expression of all markers were significantly increased with the increase of patient's age (? 50 years), and furthermore an increase in men expression when compared to women. Interestingly, all healthy tissues showed positive E-cadherin expression while they did not show any expression of Vimentin, MMP-2 and MMP-9. E-cadherin expression decreased, whereas expression of Vimentin increased according to the grade and stage of the tumor. Similarly, both MMP-2 and MMP-9 expression were increased with the progression of TCC. The current study conclude that a decrease in E-cadherin together with increased Vimentin, MMP-2 and MMP-9 are significant markers that correlate with poor prognosis of TCC.

The Prognostic Value of some Epithelial-Mesenchymal Transition Markers and Metastasis-Related Markers in Human Transitional Cell Carcinoma of the Bladder

Recent reports provided evidence that epithelial to mesenchymal transition (EMT) and some matrix metalloproteinases (MMPs) contribute to the invasion and metastasis of cancer cells. This study investigated the expression pattern of some EMT markers (E-cadherin and Vimentin) and some MMPs (MMP-2 and MMP-9) in transitional cell carcinoma (TCC). Fifty five paraffin embedded biopsies were included in this study. Expression pattern of E-cadherin and Vimentin was evaluated by immunohistochemistry while cytoplasmic mRNA expression of both MMP-2 and MMP-9 were determined by in situ hybridization. The expression of all markers were significantly increased with the increase of patient's age (? 50 years), and furthermore an increase in men expression when compared to women. Interestingly, all healthy tissues showed positive E-cadherin expression while they did not show any expression of Vimentin, MMP-2 and MMP-9. E-cadherin expression decreased, whereas expression of Vimentin increased according to the grade and stage of the tumor. Similarly, both MMP-2 and MMP-9 expression were increased with the progression of TCC. The current study conclude that a decrease in E-cadherin together with increased Vimentin, MMP-2 and MMP-9 are significant markers that correlate with poor prognosis of TCC.

The expression and clinical significance of ZBTB7 in transitional cell carcinoma of the bladder.

Zinc finger and BTB domain containing 7A (ZBTB7) is a ZBTB protein family member of transcriptional repressors that serves a critical role in cell transformation and malignancy. However, the association between ZBTB7 expression in bladder cancer tissues and the prognosis of patients remains unclear. The aim of the current study was to detect the expression of ZBTB7 in transitional cell carcinoma (TCC) of the bladder and normal bladder mucous tissues to evaluate the diagnostic and prognostic value of ZBTB7 in TCC of the bladder. A total of 100 TCC specimens were analyzed and the expression of ZBTB7 mRNA was examined via reverse transcription-quantitative polymerase chain reaction. The expression of ZBTB7 protein was examined by western blotting and immunohistochemistry. The association between ZBTB7 expression and the clinical prognosis of patients from the TCGA database was analyzed. High expression of ZBTB7 mRNA and protein in TCC tissue was detected and TCC expression was significantly higher in TCC tissue than in normal bladder mucous tissues (P<0.05). Furthermore, ZBTB7 expression was associated with recurrence, a larger tumor size and higher tumor grade. In terms of overall and recurrence-free survival, the group expressing high levels of ZBTB7 exhibited lower overall and recurrence-free survival compared with the low ZBTB7 expression group, although these differences were not statistically significant. Therefore, ZBTB7 may be important in the initiation and progression of TCC.

Targeting XBP1-mediated β-catenin expression associated with bladder cancer with newly synthetic Oridonin analogues.

Conventional chemotherapy is commonly used for advanced stages of transitional cell carcinoma (TCC) with modest success and high morbidity; however, TCC eventually develops resistance. Muscle invasive bladder cancer (MIBC) is recognized as a lethal disease due to its poor response to traditional chemotherapy. Numerous studies have implicated β-catenin, a critical effector in Wnt–mediated pathway associated with epithelial-mesenchymal transition and cancer stem cell, is involved in TCC progression, and furthermore closely associated with chemo-resistance. In this study, we discovered a novel natural product analogue CYD 6-17 that has a potent inhibitory effect on TCC cells exhibiting drug resistance to various chemotherapeutics, with an IC50 at nM range. Delivery of CYD 6-17 significantly inhibited the tumor growth using xenograft model but without detectable side effects. Mechanistically, it targeted β-catenin gene transcription by decreasing the binding of XBP1 to the promoter region, which appeared to be a new regulatory mechanism for β-catenin gene expression. Clinically, XBP1 expression correlated with the poor overall survival of patients. Overall, this study unveils unique mechanism of β-catenin gene regulation in advanced TCC and also offers a potential rational therapeutic regimen to MIBC.

MiR-130b, an onco-miRNA in bladder cancer, is directly regulated by NF-κB and sustains NF-κB activation by decreasing Cylindromatosis expression.

Persistent activation of NF-κB signaling is closely related to chronic inflammation and tumorigenesis. Commonly, NF-κB signaling is tightly controlled by multiple feedback loops and regulators, such as the deubiquitinases (DUBs). However, in cancer cells, NF-κB may override these feedbacks through special pathways and lead to the sustained activation. In the present study, we demonstrate that in transitional cell carcinoma (TCC) of bladder, miR-130b plays an oncogenesis role, it enhanced proliferation, invasion and migration of TCC cell, and was highly correlated with tumor progression. On the other hand, NF-κB directly regulated the transcription of miR-130b by binding with its promoter region. Importantly, we verify that, through deceasing the expression of Cylindromatosis (CYLD), a K63-specific DUB and endogenous blocker of NF-κB signaling, miR-130b can in return sustain the persistent activation of NF-κB, which may promote the malignant progression of TCC. Thus, the present study uncovers a potential signaling transduction in which NF-κB is continuously activated, and may provide a novel therapeutic approach for the clinical management of TCC.

Association of glutathione transferase omega polymorphisms with risk and survival in transitional cell carcinoma of urinary bladder

Exposure to chemical carcinogens, especially smoking and occupational exposure, is an etiologic factor for transitional cell carcinoma (TCC) of the urinary bladder. Cytosolic glutathione transferases (GSTs) are a superfamily of enzymes that protect normal cells by catalyzing conjugation reactions of electrophilic compounds, including carcinogens, to glutathione. Several types of allelic variations have been identified within GSTO class, with GSTO1*A140D (NCBI SNP ID: rs4925, 419 C to A) and GSTO2*N142D (NCBI SNP ID: rs156697, 424 A to G) being the most common. Individuals with the GSTO2*G/G genotype carry a higher risk for TCC. Our results indicate that GSTO1*C/GSTO2*G haplotype is associated with increased TCC risk, which is more pronounced in carriers of variant GSTO2*G/G combined with smoking. Furthermore, GSTO1*A/A and GSTO2*G/G genotypes may have a prognostic/pharmacogenomic role in patients with muscle invasive TCC. Newly discovered deglutathionylase activity of GSTO1 suggests its potential important role in redox perturbations present in TCC, which might contribute to progression of TCC.

The differential expression of EphB2 and EphB4 receptor kinases in normal bladder and in transitional cell carcinoma of the bladder.

Effective treatment of transitional cell carcinoma (TCC) of the bladder requires early diagnosis. Identifying novel molecular markers in TCC would guide the development of diagnostic and therapeutic targets. Ephrins mediate signals via tyrosine kinase activity that modulates diverse physiologic and developmental processes, and ephrins are increasingly implicated in carcinogenesis. The aim of our study was to examine the differential regulation of EphB4 and EphB2 in normal bladder and in TCC of the bladder in 40 patients undergoing radical cystectomy for curative intent. Immunostaining and Western blotting revealed that normal urothelium expresses EphB2 (20 of 24 cases, 83% of the time) not EphB4 (0 of 24 cases, 0%). In sharp contrast, TCC specimens show loss of EphB2 expression (0 of 34 cases, 0%) and gain of EphB4 expression (32 of 34, 94%). Furthermore, EphB4 signal strength statistically correlated with higher tumor stage, and trended toward the presence of carcinoma in situ (CIS). These results are confirmed by analysis of normal urothelial and tumor cell lines. EphB2 is not a survival factor in normal urothelium, while EphB4 is a survival factor in TCC. Treatment of bladder tumor xenograft with an EphB4 inhibitor sEphB4-HSA leads to 62% tumor regression and complete remission when combined with Bevacizumab. Furthermore, tissue analysis revealed that sEphB4-HSA led to increased apoptosis, decreased proliferation, and reduced vessel density, implicating direct tumor cell targeting as well as anti-angiogenesis effect. In summary loss of EphB2 and gain of EphB4 expression represents an inflection point in the development, growth and possibly progression of TCC. Therapeutic compounds targeting EphB4 have potential for diagnosing and treating TCC.

Interleukin-11, an interleukin-6-like cytokine, is a promising predictor for bladder cancer prognosis

Numerous studies have suggested that interleukin 11 (IL-11) has roles in human gastric, prostate and bone cancer and endometrial carcinoma. Hence, we evaluated the expression of IL-11 in bladder cancer and the correlation of IL-11 levels and clinico-pathological features. The expression of IL-11 in primary human bladder cell culture, human bladder cancer cell lines, transitional cell carcinoma (TCC) and non-cancerous bladder tissues (NATs) were analyzed by western blotting. Enzyme-linked immunosorbent assay (ELISA) for urinary IL-11 was performed to compare the IL-11 levels in healthy subjects and subjects diagnosed with bladder cancer. Our study suggested that the expression of IL-11 in human bladder cancer cell lines and TCC was downregulated compared with primary human bladder cell culture and matched NATs. We also demonstrated reduced urinary levels of IL-11 in subjects with bladder cancer compared with healthy subjects. Furthermore, we revealed that the levels of IL-11 were associated with tumor grade and stage. The results suggested that reduced levels of IL-11 may play an important role in the carcinogenesis and progression of TCC. They also indicated that IL-11 may be a promising predictor for prognosis of TCC.

Relationships among MTHFR a1298c Gene Polymorphisms and Methylation Status of Dact1 Gene in Transitional Cell Carcinomas

The purpose of this study was to determine the relationship between methylation status of the Dact1 gene and MTHFR a1298c polymorphic forms in transitional cell carcinoma tissues in a Chinese population. Polymorphisms of folate metabolism enzyme gene MTHFR were assessed by restrictive fragment length polymorphism (RFLP) methods and PCR-based DNA methylation analysis was used to determine the CpG island methylation status of the Dact1 gene. Associations between the methylation status of the Dact1 gene and clinical characteristics, as well as MTHFR a1298c polymorphisms, were analyzed. aberrant methylation of the Dact1 gene was found in 68.3% of cancer tissues and 12.4% of normal tissues,. The methylation rate of the Dact1 gene in cancer tissues was significantly higher in patients with lymph node metastasis than in those without lymph node metastasis (46.3% vs. 17.2%, P = 0.018). No association was found between aberrant DNA methylation and selected factors including sex, age, tobacco smoking, alcohol consumption and green tea consumption. After adjusting for potential confounding variables, variant allele of MTHFR a1298c was found to be associated with methylation of the Dact1 gene. Compared with wild type CC, the odds ratio was 4.33 (95% CI: 1.06-10.59) for AC and 4.95 (95% CI: 1.18-12.74) for AA. The N stage in TNM staging and the occurrence of lymph node metastasis were associated with an MTHFR 1298 AAμAC genotype (P<0.05). MTHFR 1298 AC and AA genotypes might help maintain a normal methylation status of the Dact1 gene, aberrant CpG island methylation of which is closely related to the genesis and progression of transitional cell carcinoma.

The Expression of Pigment Epithelium-Derived Factor in Bladder Transitional Cell Carcinoma

BackgroundPigment epithelium-derived factor (PEDF) is an anti-angiogenic factor. The purpose of this study is to examine the involvement of PEDF in the angiogenesis and biological behavior of bladder transitional cell carcinoma (TCC).MethodsWe examined the expression of PEDF in 99 bladder TCCs and ten non-neoplastic tissues, and evaluated microvessel density (MVD).ResultsThe positive immunoreactivity for PEDF was seen in normal urothelium in 60% (6/10) and TCC in 13% (13/99). The PEDF expression had a significant correlation with MVD, i.e., a low MVD in 42% (5/12), a middle MVD in 11% (8/76) and a high MVD 0% (0/11) of tumors. The PEDF expression was not significantly correlated with the differentiation and invasion of TCC, but the degree of MVD was significantly higher in both high grade TCC and the pT2 tumors.ConclusionsThe degree of PEDF expression is significantly higher in normal bladder urothelium than bladder TCC; it is inversely correlated with the angiogenesis; and it is not related to the differentiation and progression of TCC. It can therefore be concluded that bladder TCC would initially occur if there is a lack of the PEDF expression.

Genetic Variability in 8q24 Confers Susceptibility to Urothelial Carcinoma of the Upper Urinary Tract and is Linked With Patterns of Disease Aggressiveness at Diagnosis

Upper urinary tract urothelial carcinoma is rare. Evidence shows that it behaves differently from urothelial bladder tumors. A polymorphism located at the T allele of rs9642880 on chromosome 8q24 is linked to an enhanced risk of bladder tumors. We explored the association of this polymorphism with susceptibility to upper urinary tract urothelial carcinoma. We genotyped the constitutional DNA of 261 patients with upper urinary tract urothelial carcinoma and 261 healthy controls matched for age, gender, smoking habit and ethnicity. Polymorphisms at rs9642880 on chromosome 8q24 were determined using the 5' nuclease polymerase chain reaction method with specific primers and probes. Frequencies were compared between cases and controls. Genotypes were in Hardy-Weinberg equilibrium for cases and controls. Mean patient age was 68.7 years. The T/T genotype resulted in a significantly higher risk of upper urinary tract urothelial carcinoma (OR 1.72, 95% CI 1.1-2.8, p = 0.028). Using single polytomous regression analysis the T/T genotype was also associated with aggressive tumors when stratified by stage (p = 0.003), or grade G2 (p = 0.04) or G3 (p = 0.01). Our results strongly suggest that the T/T rs9642880 genotype is a risk factor for upper urinary tract urothelial carcinoma, as previously shown for bladder tumors. In contrast to bladder carcinoma, for upper urinary tract urothelial carcinoma the T/T genotype is associated with aggressiveness.

Abstract 4187: Phospho-Akt status as a predictor of disease free interval in patients with muscle invasive transitional cell carcinoma of the bladder

Abstract Background: Akt is an ubiquitous signaling molecule that is associated with a wide array of biological effects including cell growth and proliferation. The dysregulation of Akt has been associated with cancer cell transformation, invasion and metastasis. While the roles of mutations in FGFR3, p53 and Ras have been the focus of the majority of studies of Transitional cell carcinoma (TCC) of the bladder, recent evidence suggests a link between the Akt pathway and the progression of TCC. Mutations in Akt pathway members have been identified in subsets of bladder TCCs, and Akt activation in TCC cell lines confers resistance to chemotherapeutics and promotes invasion in vitro. This led us to hypothesize that activated Akt may be an important biomarker in the progression of muscle invasive TCC. Methods: Tissue was obtained from 67 patients who had undergone radical cystectomy procedures for treatment of muscle invasive TCC. Tissue sections were taken from paraffin embedded samples and subjected to immunohistochemical analysis with anti Akt antibody to detect the activated form of Akt. A database of patient demographics, disease, treatment and survival parameters was generated and used to correlate 4, 12, 24 and 36 month disease-free intervals (DFI). Results: 53 of the 67 patients had sufficient follow-up to allow for DFI analysis. Of these 53 patients, 48 stained positively for pAkt, while 5 stained negatively. None of the pAkt negative patients had recurrent disease, while the pAkt negative patients suffered cumulative recurrence in 10, 19, 24, and 25 of the 48 patients at 4, 12, 24 and 36 months respectively. Chi square analysis reveals statistically significant differences in recurrence rates between pAkt negative and negative patients at 24 (p=0.0235) and 36 months (p=0.016). Tumor stage and nodal status were also strong predictors of recurrence. Conclusions: pAkt negative patients were found to have a significantly better prognosis for TCC recurrence, which indicates that pAkt status could be useful in helping to determine the prognosis for patients with muscle invasive TCC. These results lead us to speculate that the use of a serine threonine kinase inhibitor may prove to be therapeutically beneficial for TCC patients undergoing concurrent chemotherapy for their disease. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4187. doi:10.1158/1538-7445.AM2011-4187

Correlation between the cyclooxygenase-2 expression and tumor interstitial vascular density in transitional cell carcinoma of urinary bladder

Objective To investigate the role of cyclooxygenase-2 (COX-2) and microvascular density (MVD) in the tumorigenesis and progression of transitional cell carcinoma of urinary bladder (BTCC), and their clinical significance. Methods Immunohistochemistry was used to study COX-2 and the tumor interstitial vascular density in 103 cases of BTCC, 13 cases of normal bladder tissues and 21 cases of chronic cystitis tissues. Results The COX-2 positive expression rate in BTCC was 70.9% (73/103), and that in chronic cystitis and normal bladder tissues was negative. The expression of COX-2 in BTCC was positively correlated to the tumor interstitial vascular density. Both of them were related to the tumor pathologic grade, the invasion of the tumor tissue, the lymphatic metastasis and recurrence of tumor ( P 0. 05 ). Conclusion COX-2 was overexpressed in BTCC and played an important role in oncogenesis, development and recurrence of BTCC.The expression of COX-2 was positively related to the tumor vascularization. Key words: Bladder neoplasms; Cyclooxygenase

Expression of Fas in peripheral blood T lymphocytes in patients with bladder urothelial carcinom

Objective To investigate the relationship between the Fas expression in peripheral blood T cells and bladder transitional cell carcinoma (TCC). Methods The Fas expression in peripheral blood T cells of 52 patients with TCC and 37 healthy people was detected by flowcytometry. The Fas-posi-tive rate was compared by t test between two designed groups. Results The Fas expression in CD4~+ [(20.74±9.02)%] and CD8~+[ (7.51±5.93 )% ] was increased in TCC patients as compared with controls ( P 0.05 ). Conclusion The abnormal Fas expression in peripheral blood T cells may play an important role in the development and progression of TCC. Key words: Bladder neoplasm; Fas protein; Lymphocyte; Apoptosis

Comparison of distributions of survivin among tissues from urinary bladders of dogs with cystitis, transitional cell carcinoma, or histologically normal urinary bladders

To compare distributions of survivin among tissues from urinary bladders of dogs with cystitis, transitional cell carcinoma (TCC), or histologically normal urinary bladders. 24 archived and 7 fresh-frozen specimens of urinary bladders from dogs with cystitis. Immunohistochemical analysis of archived tissue specimens was performed to identify survivin protein in the nucleus and cytoplasm of cells by use of polyclonal rabbit anti-survivin antibody. Tissues that contained > or = 5% immunoreactive cells were considered positive for survivin protein. Reverse-transcription PCR analysis was performed on fresh-frozen tissues to identify survivin mRNA. Data on tissues from dogs with TCC or histologically normal urinary bladders that were obtained during another study were used for statistical comparisons. Twelve of 24 (50%) cystitic tissues were positive for nuclear survivin, compared with 28 of 41 (68%) TCC tissues and 0 of 46 (0%) normal tissues. Two of 24 (8%) cystitic tissues were positive for cytoplasmic survivin, compared with 7 of 41 (17%) TCC tissues and 17 of 46 (37%) normal tissues. Proportions of specimens that contained nuclear or cytoplasmic survivin were significantly different between cystitic and normal tissues but not between cystitic and TCC tissues. Four of 7 cystitic tissues were positive for survivin mRNA, which was comparable with results for TCC and normal tissues. Nuclear survivin was detected in TCC and cystitic tissues but not in normal urinary bladder tissues. Additional studies are needed to determine whether nuclear survivin contributes to the development or progression of TCC.

Prognostic significance of HIF‐1α polymorphisms in transitional cell carcinoma of the bladder

Recently, two single nucleotide polymorphisms in the hypoxia-inducible factor-1 alpha (HIF-1 alpha) gene, P582S and A588T, were shown to cause significantly higher transcriptional activity than the wild type. We investigated the association between the HIF-1 alpha polymorphisms and the incidence and progression of transitional cell carcinoma of the bladder, and the relationship between the polymorphisms and the tissue vascular endothelial growth factor (VEGF) level or microvessel density (MVD). A total of 219 patients with bladder cancer and 464 healthy native Japanese control subjects were enrolled. Tissue VEGF and HIF-1 alpha expression levels and the mean MVD were evaluated in 73 radical cystectomy specimens by immunohistochemistry. The HIF-1 alpha genotype did not significantly influence the incidence or disease status of bladder cancer. Among patients who underwent radical cystectomy, those with a variant allele had significantly worse disease-free survival (p = 0.001) and disease-specific survival (p = 0.006) than those without a variant allele. Multivariate analysis using a Cox proportional hazard model revealed that the presence of a variant allele was an independent predictor of disease-free survival (HR = 3.10, 95%CI = 1.38-6.99, p = 0.006). Although not statistically significant, the moderate/high expression levels of VEGF in tumor tissues were more frequently observed in patients with a HIF-1 alpha variant allele (11/13, 84.6%) than in those without (33/60, 55%, p = 0.063). The HIF-1 alpha polymorphisms may have a significant influence on the poor prognosis of the patients undergoing radical cystectomy for bladder cancer, while they seem to have no relation to the bladder cancer occurrence.

Expression of LAT1 predicts risk of progression of transitional cell carcinoma of the upper urinary tract

L-type amino acid transporter 1 (LAT1), a neutral amino acid transporter, requires covalent association with the heavy chain of 4F2 cell surface antigen (4F2hc) for its functional form. We investigated the importance of LAT1 and 4F2hc expressions to progression in upper urinary tract cancer. We examined their expressions and their relationships to clinicopathologic parameters and clinical outcome in 124 cases. Positive expressions of LAT1 (protein and messenger ribonucleic acid) and 4F2hc (protein) were recognized in 79.8, 89.5, and 87.9% of tumor samples, respectively. In tumor cells, LAT1 protein was detected either as nodular granules within the cytoplasm or diffusely within the cytoplasm and/or on plasma membrane. In the normal urothelium, its expression was detected as nodular granules within the cytoplasm. A correlation with stage was shown for LAT1 protein expression and for a cooperative expression of LAT1 protein with 4F2hc protein (active form of LAT1 protein). Further, in all tumors, a cooperative expression of LAT1 protein and 4F2hc protein was significantly correlated with both overall and disease-free survival rates in the univariate analysis but not in the multivariate analysis. In conclusion, the detection of the active form of LAT1 protein would appear to be of value in informing the risk of progression in transitional cell carcinoma of the upper urinary tract.

Promoter hypermethylation of the 14-3-3 σ, SYK and CAGE-1 genes is related to the various phenotypes of urinary bladder carcinomas and associated with progression of transitional cell carcinomas

To explore the significance of epigenetic mechanisms in urinary bladder carcinogenesis mediated by methylation of cytosine in CpG dinucleotides at 5' promoter regions, we analysed the methylation status of a broad panel of different genes in transitional cell carcinomas (TCC) and nonurothelial cancers, among which the 14-3-3 sigma, SYK and CAGE-1 genes were recognised as promising target genes. Using methylation-specific PCR, the rate of DNA hypermethylation proved to be related to the various histopathological cancer subtypes. The higher frequency of promoter methylation of the 14-3-3 sigma (57.1%) and SYK (64.3%) genes in high-grade, high-stage TCC in association with a reduced or even lacking immunohistochemical protein expression than in low-grade, low-stage TCC (28.6% and 42.9%, respectively), indicates that aberrant methylation of these genes plays an essential role in the progression of TCC. The importance of DNA hypermethylation in the conversion of TCC from a low to a high malignant potential was strongly supported by the finding that, unlike superficial low-grade TCC, advanced muscle invasive TCC showed a concurrent promoter methylation of the 14-3-3 sigma, SYK and CAGE-1 genes. Squamous cell carcinomas revealed a peak incidence of hypermethylation of the 14-3-3 sigma gene (80%), and conversely, the lowest methylation frequency of the SYK gene (13.3%). Undifferentiated small cell carcinomas disclosed a promoter methylation of the 14-3-3 sigma, SYK and CAGE-1 genes in only a quarter each for the cases. Although a correlation between the methylation status and gene activity in squamous cell and undifferentiated small cell carcinomas was not observed, the underexpression of the SYK protein products in both cancer types and additionally of the 14-3-3 sigma protein in small cell carcinomas appeared to be related to the aggressive clinical behaviour of both these nonurothelial bladder carcinomas. The relevance of the high frequency of DNA hypermethylation of the CAGE-1 antigen in TCC and squamous cell carcinomas merits further study, particularly in relation to anticancer immunotherapy. The methylation status of the PTEN, COX-2, RUNX-3 and HIC-1 genes was found to be unaltered. In conclusion, the different patterns of aberrant methylation of the 14-3-3 sigma, SYK and CAGE-1 genes in the various histopathological cancer types of the urinary bladder point to a role in tumor cell differentiation, resulting in the phenotypical conversion of TCC into nonurothelial carcinomas and in the progression of TCC to a more malignant potential.

Multifocal Urothelial Cancers With the Mutator Phenotype are of Monoclonal Origin and Require Panurothelial Treatment for Tumor Clearance

UC is a disease of the entire urothelium, characterized by multiplicity and multifocality. The clonal relationship among multiple UCs has implications regarding adjuvant chemotherapy. It has been investigated in studies of chromosomal alteration and single gene mutation. However, these genetic changes can occur in unrelated tumors under similar carcinogenic selection pressures. Tumors with high MSI have numerous DNA mutations, of which many provide no selection benefit. While these tumors represent an ideal model for studying UC clonality, their low frequency has prevented their previous investigation. We investigated 32 upper and lower urinary tract UCs with high MSI and 4 nonUC primary cancers in 9 patients. We used the high frequency and specificity of individual DNA mutations in these tumors (MSI at 17 loci) and the early timing of epigenetic events (methylation of 7 gene promoters) to investigate tumor clonality. Molecular alterations varied among tumors from different primary organs but they appeared related in the UCs of all 9 patients. While 7 patients had a high degree of concordance among UCs, in 2 the UCs shared only a few similar alterations. Genetic and epigenetic abnormalities were frequently found in normal urothelial samples. Multiple UCs in each patient appeared to arise from a single clone. The molecular order of tumor development varied from the timing of clinical presentation and suggested that residual malignant cells persist in the urinary tract despite apparent curative surgery. These cells lead to subsequent tumor relapse and new methods are required to detect and eradicate them.