Transitional Cell Carcinoma Tissues Research Articles

Pattern of Occurrence and Management of Dual Malignancy: Our Institutional Experience

Background: Management of dual malignancies is challenging in the present scenario of advanced techniques and increased life expectancy. Objectives: To determine the demographic and clinical profile of patients with dual malignancies and the management received. Methods: In this retrospective analysis of the data of patients presenting with histologically proven synchronous or metachronous dual malignancy, we evaluated the demographic and clinical profile of patients with dual malignancies and the management received. Results: In this study, 158 cases with multiple primary cancers, of whom 57 were synchronous and 101 were metachronous, were observed from January 2017 to December 2021. The maximum period for synchronous tumor occurrence was seen at 6 months (180 days). The interval of occurrence of metachronous tumors ranged from 1 to 15 years, with an average of 4.8 years for the entire group. Moreover, 73 (46.2%) were females, and 85 (53.8%) were males, with a male-to-female ratio of 1.2: 1. The most common site of primary tumor with dual malignancies was the head and neck (31%), followed by breast cancers (21%). The most frequent pathologic type was adenocarcinoma (40%), followed by squamous carcinoma (38%), hematopoietic and lymphoid tissues (7.5%), transitional cell carcinoma (6.3%), sarcomas and soft tissue tumors (2.5%). Conclusions: Even in complete clinical remission, every cancer patient must take into account the possibility of developing a second malignancy and must be closely monitored.

Frequent expression of a novel cancer testis antigen, protein kinase human monopolar spindle 1 (hMps1/TTK) in human urinary bladder transitional cell carcinoma.

Urothelial bladder cancer (UBC) is a frequently occurring malignancy of the urinary tract. The present study was undertaken to evaluate the mRNA and immunohistochemical (IHC) expression of protein kinase human monopolar spindle 1 (hMps1/TTK) gene in transitional cell carcinoma (TCC) of the bladder and correlate its expression with the clinicopathological characteristics of patients. In the present study, quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) was used to evaluate TTK mRNA expression in TCC. IHC analysis of TTK was also evaluated. Independent Student's t, ANOVA and chi-square (χ2) tests were used to analyze the data statistically. The frequency of TTK mRNA over expression was detected in 50% of UBC (38/76) by qRT-PCR. Relative mean fold expression of TTK mRNA was found significantly (p < 0.05) higher in muscle-invasive bladder cancer (MIBC) as compared to non-muscle-invasive bladder cancer (NMIBC) patients (8.96 ± 4.51 vs. 5.64 ± 3.53, p = 0.03). Moreover, IHC reveals heterogenous immunostaining pattern of TTK in TCC tissues. The frequency of TTK protein over expression was detected in 56.9% (37 of 65) UBC patients. No significant IHC expression of TTK was detected among adjacent noncancerous tissues (ANCTs) and benign prostatic hyperplasia (BPH) used as control. Collectively our study observations conclude that TTK is a novel cancer/testis antigen (CTA) as a diagnostic marker for early diagnosis of UBC.

Biochemical study of E-Cadherin and SATB1 in Urothelial Bladder Carcinoma of Egyptian Patients

Bladder cancer was considered the third most common tumor in Egypt according to the National Cancer Institute (NCI) registry. However, Transitional cell carcinoma of the urinary bladder represents about 90% of all bladder cancer cases. Method; The aim of the study was to evaluate the expression levels (mRNA and protein) of E-cadherin and special AT-rich sequence binding protein-1 (SATB1) as biomarkers for transitional cell carcinoma (TCC) using immunohistochemistry and quantitative reverse transcription PCR (RT-qPCR) techniques of the Egyptian urinary bladder patients compared to the noncancerous bladder tissues. The study included 48 TCC tissues (41 males and 7 females) of different grades and stages and 12 noncancerous bladder tissues (7 males and 5 females). Results; A statistically significant association regarding smoking and gender were found between the bladder transitional cell carcinoma (BTCC) group and the control group. However, there was no statistically significant difference in comparing the presence of schistosomiasis in both studied groups. The expression of E-cadherin was significantly down expressed and SATB1 was significantly over expressed in TCC tissues compared to the noncancerous bladder tissues at both mRNA and protein levels. Decreased expression of E-cadherin and over expression of SATB1 were statistically associated with tumor stage, tumor grade, and tumor size. On the other hand, no statistically significant difference was detected between the expressions of the two studied genes and either gender or age. A significant association was found between the protein expressions of E-cadherin and SATB1. Pearson correlation test showed significantly negative correlation between mRNA expressions of E-cadherin and SATB1. Conclusion; E-cadherin and SATB1 expressions may be considered as good biochemical markers for the diagnosis and prognosis of transitional cell carcinoma in urinary bladder of Egyptian patients.

Increased expression of POLR3G predicts poor prognosis in transitional cell carcinoma.

BackgroundPrevious studies have shown that RNA Polymerase III Subunit G (POLR3G) has oncogenic effects in cultured cells and mice. However, the role of POLR3G in transitional cell carcinoma (TCC) has not been reported. This study explores the potential of POLR3G as a novel molecular marker for TCC.MethodsThe RNA sequencing data and clinical information of patients with TCC were downloaded from The Cancer Genome Atlas official website. Transcriptome analysis was performed as implemented in the edgeR package to explore whether POLR3G was up-regulated in TCC tissues compared to normal bladder tissues. The expression of POLR3G in bladder cancer cell line T24 and human uroepithelial cell line SV-HUC-1 were detected via quantitative real time polymerase chain reaction (qRT-PCR). Correlations between POLR3G expression and clinicopathological characteristics were analyzed using Mann-Whitney U test or Kruskal-Wallis H test. Clinicopathological characteristics associated with overall survival were explored using the Kaplan-Meier method and Cox regression analyses. Gene set enrichment analysis (GSEA) was performed to explore the associated gene sets enriched in different POLR3G expression phenotypes and the online tool Tumor IMmune Estimation Resource (TIMER) was used to explore the correlation between POLR3G expression and tumor immune infiltration in TCC.ResultsTranscriptome analysis showed that POLR3G was significantly up-regulated in TCC tissues compared to normal bladder tissues. Furthermore, qRT-PCR revealed high expression of POLR3G in T24 cells compared to SV-HUC-1 cells. Overall, POLR3G expression was associated with race, tumor status, tumor subtype, T classification, and pathological stage. Kaplan-Meier survival analysis revealed that higher POLR3G expression was associated with lower overall survival. The univariate Cox regression model revealed that age at diagnosis, pathological stage, and POLR3G expression were associated with prognosis of TCC patients. Further multivariate analyses identified these three clinicopathological characteristics as independent prognostic factors for overall survival. GSEA analysis showed that several gene sets associated with tumor development and metastasis, including TGF-β signaling, PI3K-AKT-mTOR signaling, and IL6-JAK-STAT3 signaling, were significantly enriched in POLR3G high expression phenotype. Immune infiltration analysis revealed that the expression of POLR3G was significantly correlated with infiltrating levels of immune cells, including CD8+ T cells, neutrophils, and dendritic cells; and the expression of POLR3G was also significantly correlated with the expression of immune checkpoint molecules, such as PD1, PD-L1, PD-L2, CTLA4, LAG3, HAVCR2, and TIGIT.ConclusionsPOLR3G was up-regulated in TCC and high POLR3G expression correlated with poor prognosis. POLR3G can potentially be used as a prognostic marker for TCC and might be of great value in predicting the response to immunotherapy.

A functional variant rs1464938 in the promoter of fibroblast growth factor 12 is associated with an increased risk of bladder transitional cell carcinoma

Increasing evidence shows that inflammation plays critical roles in the tumorigenesis of bladder cancer. Fibroblast growth factor 12 (FGF12), a kind of inflammatory cytokine, is located in the region of 3q28 that has been demonstrated to be a bladder cancer risk locus by genome wide association study (GWAS). In this study, we aimed to investigate the association of GWAS signal rs710521 and rs884309 and rs1464938 in the promoter of FGF12 with the risk of bladder transitional cell carcinoma (TCC). The polymorphisms were analyzed by using a Taqman assay in 331 TCC patients and 516 age-, gender-, and ethnicity-matched controls. The expression levels of FGF12 mRNA were examined in TCC and non-cancerous normal tissues by using quantitative real-time PCR and the luciferase activity was determined by using the Dual-Luciferase Assay System. The rs1464938 AA genotype and A allele were associated with a significantly increased risk of TCC (AA vs. GG: adjusted OR = 2.54, 95% CI, 1.49–4.35, P < 0.001; AA vs. AG/GG: adjusted OR = 2.25, 95% CI, 1.36–3.71, P = 0.002; A vs. G: adjusted OR = 1.44, 95% CI, 1.15–1.80, P = 0.001, respectively). Haplotype analysis showed that rs884309G- rs1464938A haplotype was associated with an increased risk of TCC (OR = 1.61, 95% CI, 1.23–2.11, P = 0.001). Functional analysis showed that the rs1464938 AG/AA genotypes exhibited higher levels of FGF12 mRNA in TCC tissues and the rs1464938 A allele enhanced FGF12 promoter activity (P < 0.05). These findings suggest that the rs1464938 A allele at the 3q28 locus contribute to the development of TCC by regulating FGF12 expression levels.

Gene expression of prostaglandin EP4 receptor in three canine carcinomas

BackgroundChronic inflammation mediated by the cyclooxygenase enzymes, specifically their product prostaglandin E2 (PGE2), can result in the development of cancer. PGE2 promotes cell proliferation, apoptosis, and angiogenesis through interaction with its specific receptors (EP1 receptor - EP4 receptor [EP1R-EP4R]). In multiple human cancers, the expression of EP4R is associated with the development of malignancy and a poor prognosis. The expression of EP4R has not yet been evaluated in canine tumors. The aim of this study was to characterize the mRNA gene expression of EP4R (ptger4) in canine squamous cell carcinoma (SCC), apocrine gland anal sac adenocarcinoma (AGASACA), and transitional cell carcinoma (TCC). Archived tumor samples of canine cutaneous SCC (n = 9), AGASACA (n = 9), and TCC (n = 9), and matched archived normal tissue controls were evaluated for mRNA expression of canine EP4R using RNA in situ hybridization (RNAscope®). Quantification of RNAscope® signals in tissue sections was completed with an advanced digital pathology image analysis system (HALO). Data was expressed as copy number, H-index, and percent tumor cell expression of EP4R.ResultsIn all canine SCC, AGASACA, and TCC samples evaluated, strong universal positive expression of EP4R was identified. For SCC and AGASACA, mRNA EP4R expression was statistically higher than that of their respective normal tissues. The TCC tissues displayed significantly less mRNA EP4R expression when compared to normal bladder mucosa.ConclusionsThese results confirm the mRNA expression of canine EP4R in all tumor types evaluated, with SCC and AGASACA displaying the highest expression, and TCC displaying the lowest expression. This study also represents the first reported veterinary evaluation of EP4R expression using the novel in situ hybridization technique, RNAscope®.

The expression of apolipoproteina1 and its correlation with infiltration of urologic neoplasm.

BackgroundTo explore the differential expression of apolipoproteinA1 (APOA1) in urologic neoplasm patient compared with controls, as well as investigates whether APOA1 correlated with infiltration of urologic neoplasm.MethodsA total of 59 tissue sections of surgically-resected urologic neoplasm and 6 cases of normal tissue sections were collected. Fourteen cases of urine samples from transitional cell carcinoma patients and 6 cases urine samples from controls were also applied in this experiment. We also selected 6 cases of fresh bladder transitional cell carcinoma tissues. The urologic neoplasm tissue sections were classified into infiltration and non-infiltration urologic neoplasm groups. The expressions of APOA1 between urologic neoplasm and normal control were detected by Western blot, Immunohistochemistry and qRT-PCR. The method of Immunohistochemistry was applied to examine the differences of APOA1 expression between infiltration and non-infiltration urologic neoplasm tissue section groups.ResultsCompared with none expression in normal controls, APOA1 was exhibited higher level in urologic neoplasm patient’s urine and fresh bladder transitional cell carcinoma tissues (P<0.05). There were statistical differences of APOA1 between infiltration and non-infiltration urologic neoplasm tissue section groups. APOA1 expressions were found to be up-regulated in the infiltration neoplasm tissue sections compared to non-infiltration group (P<0.001).ConclusionsAPOA1 could act as a valuable biomarker for predicting the occurrence and development of urologic neoplasm.

336 Increased Nicotinamide Phosphoribosyltransferase and Cystathionine B-Synthase in Renal Oncocytomas, Renal Transitional Cell Carcinoma, and Renal Cell Carcinoma

Renal cell carcinoma (RCC) and transitional cell carcinoma (TCC) are the two most common renal malignancies, while renal oncocytomas are common benign renal neoplasms. Nicotinamide phosphoribosyltransferase (Nampt) catalyzes the rate-limiting step in nicotinamide adenine dinucleotide synthesis, while cystathionine B-synthase (CBS) catalyzes H2S synthesis. Both Nampt and CBS show concomitant increased expression in some tumors, with their expression positively correlating with higher tumor grade. We employed tissue microarray studies to analyze Nampt and CBS protein levels in benign renal cortex, RCC, TCC, and renal oncocytoma tissue samples. For Nampt, 29 benign, 49 TCCs, 94 RCCs, and six oncocytomas were examined. For CBS, 11 benign, 53 TCCs, 101 RCCs, and six oncocytomas were examined. The expression of both proteins was increased in TCC, RCC, and oncocytomas, compared to benign renal tissue. In RCC, the expression of both enzymes increased with increasing Fuhrman grade. To our knowledge, this is the first demonstration that Nampt and CBS are increased in renal RCC, TCC, and oncocytomas, and also increased with increasing RCC grade. Both enzymes promote tumor growth, survival, and angiogenesis in several malignancies. Increased expression of either enzyme in any oncocytoma type has not been previously reported. While preliminary, our data indicate that increased in Nampt and CBS protein expression, and possibly changes in H2S metabolism may have a role in benign and malignant renal neoplasia.

Cancer-Testis Antigens as New Candidate Diagnostic Biomarkers for Transitional Cell Carcinoma of Bladder.

To evaluate the diagnostic potential of 23 candidate genes, belonging to a category of tumor-specific antigens known as cancer-testis antigens (CTAs), in transitional cell carcinoma (TCC) patients. The expression of 16 known candidate CTAs and seven testis restricted/selective genes, predominantly expressed in the testis, was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR). Urinary exfoliated cells (UECs) and cancerous tissues of 73 TCC patients were used as cases, while 25 tumor-free adjacent bladder tissue specimens along with bladder tissue specimens and UECs of five non-TCC individuals were analyzed as controls. Among the known CTAs only MAGEA3, MAGEB4, TSGA10, PIWIL2, OIP5, and ODF4 were expressed specifically in TCC tissues and UEC samples. ACTL7A, AURKC, and CGB2 were testis-restricted/selective genes that indicated specific expression in cases in comparison to controls. MAGEA3, MAGEB4, and ODF4 mRNA was detectable in more than 50% of both TCC tissues, and UEC samples. Slight differences were detected in the mRNA expression pattern of candidate genes between the UEC samples and tumor tissues. Different panels formed by combinations of these genes can show up to 95.9% and 94.5% of positivity in TCC tissues and UEC samples, respectively, suggesting their diagnostic and surveillance potential. Meanwhile the RT-PCR assay of at least MAGEA3, MAGEB4, and ODF4 may be particularly useful for diagnostic and surveillance of TCC in the form of a multi-biomarker panel.

Selection of reference genes for gene expression studies in human bladder cancer using SYBR-Green quantitative polymerase chain reaction.

Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is a rapid, reliable and widely used method of studying gene expression profiles that requires appropriate normalization for accurate and reliable results. Reference genes are usually used to normalize mRNA levels; however, the expression levels of these reference genes may vary between cell types, developmental stages, species and experimental conditions. Therefore, a normalization strategy is an important precondition for reliable conclusions, with endogenous controls requiring determination for every experimental system. In the present study, 18 reference genes used in various prior studies were analyzed to determine their applicability in bladder cancer. A total of 35 matched malignant and non-malignant bladder cancer (specifically transitional cell carcinoma) tissue specimens were examined. RNA and cDNA quality was stringently controlled. Candidate reference genes were assessed using SYBR-Green RT-qPCR. mRNA abundance was compared and reference genes with distinct ranges of expression to possible target genes were excluded. Genes that were differentially expressed in matched non-cancerous and cancerous samples were also excluded, using quantification cycle analysis. Subsequently, the stability of the selected reference genes was analyzed using three different methods: geNorm, NormFinder and BestKeeper. The rarely used ribosomal protein S23 (RPS23) was the most stable single reference gene, with RPS23, tumor protein, translationally controlled 1 and RPS13 comprising the optimal reference gene set for all the bladder samples. These stable reference genes should be employed in normalization and quantification of transcript levels in future expression studies of bladder cancer-associated genes.

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.

Density of tumor-infiltrating granzyme B-positive cells predicts favorable prognosis in dogs with transitional cell carcinoma

Although tumor-infiltrating lymphocytes (TILs) play a key role in anti-tumor immunity, their involvement in canine transitional cell carcinoma (TCC) is not well-documented. The objective of this study was to investigate the association between TIL number and prognosis in dogs with urinary bladder TCC. Immunohistochemical analysis of CD3 and granzyme B was performed using canine TCC (n=32) and normal bladder (n=10) tissues. The numbers of CD3+ and granzyme B+ cells located in peritumoral stroma of canine TCC were significantly higher than those in normal controls. In TCC cases, the number of CD3+ TILs was not significantly related to prognosis, whereas the abundant granzyme B+ TILs were associated with favorable outcome. Since granzyme B+ TILs were not associated with the tumor stage, the presence of granzyme B+ TILs may be an independent prognostic factor. These results suggest that granzyme B+ TILs play a role in anti-tumor immunity and inhibit tumor progression in canine TCC.

Evaluation of androgen receptor and HER2/neu receptor in tumor tissue of transitional cell carcinoma of urinary bladder and normal bladder mucosa

Background: Transitional cell carcinoma (TCC) is the most prevalent, singly accounting for 90% of all bladder cancer cases. Various markers involved in cell cycle but none of them proven its capacity as a prognostic indicator. The discovery of new biological markers may lead to early clinical prediction and may help in individualization of therapeutic approach in management of bladder cancer patients. The human epidermal growth factor receptors (EGFRs) – HER2/neu play a role in neoplastic cell growth and its role proven in breast and ovarian cancer while its role in bladder cancer still not explored its potential. Bladder cancer incidence is higher in male as compared to female. This gender related difference is still not clear in humans while it is proven in animal studies. In this study, we investigated the potential role and association of HER2/neu and androgen receptors (ARs) in etiology and prognosis of TCC. Patients and Methods: The study included 20 histopathologically confirmed cases of TCC of urinary bladder attending the Department of General Surgery, KGMU, Lucknow and 20 healthy individuals were taken as control. Immunohistochemical analysis for AR and HER2/neu receptor was done using paraffin-embedded tissue blocks treated with monoclonal mouse anti-human antibodies against AR and HER2/neu receptor, respectively. Results: HER2/neu over expression was observed in 14 (73%) of which 12 were males and 2 were females. Among these 16.7%, 33.3%, and 37.5% cases were in Grade I, II, and III, respectively. The AR overexpression was observed in 4 (20%) of which all were males. About 16.7% and 27.3% cases were in Grade II and III, respectively. Peak incidence was observed in the age group of 50–70 years (75%). Conclusions: We conclude that HER2/neu and AR can be considered as bad prognostic factor in TCC. These results suggested that targeting AR and HER2/neu may provide novel chemopreventive and therapeutic approaches for the management of bladder cancer.

Wild-type p53-induced phosphatase 1 is a prognostic marker and therapeutic target in bladder transitional cell carcinoma

Wild-type p53-induced phosphatase (Wip1) is an established oncogene and is associated with development of multiple forms of human cancer. However, the expression and role of Wip1 in human bladder transitional cell carcinoma (TCC) remains to be elucidated. In the present study, immunohistochemistry demonstrated that Wip1 was overexpressed in bladder TCC tissues compared with corresponding normal bladder tissues in 106 bladder TCC cases (P<0.0001). Furthermore, high expression levels of Wip1 were significantly associated with increasing tumor size (P=0.002), pathological grade (P=0.025), clinical T stage (P=0.001) and lymph nodal metastasis (P=0.003). Kaplan-Meier survival analysis identified that patients with high Wip1 expression levels exhibited a lower overall survival time (P<0.0001), and Cox proportional hazards regression model analysis demonstrated that Wip1 expression was an independent prognostic factor in patients with bladder TCC (P=0.025). In addition, downregulation of Wip1 expression by transfection with small interfering RNA in bladder cancer cells inhibited cell proliferation, invasion and migration (P<0.05), along with the upregulation of p53 protein levels (P<0.05). These findings suggest that Wip1 may function as a potential prognostic marker and therapeutic target in bladder cancer.

AB112. Expression of brain-specific angiogenesis inhibitor 1 and association with p53, microvessel density and vascular endothelial growth factor in the tissue of human bladder transitional cell carcinoma

ObjectiveBrain-specific angiogenesis inhibitor 1 (BAI1) was initially described in 1997, and there have since been a number of studies on its expression in different types of cancer. The aim of the present study was to investigate the expression levels of BAI1 in bladder transitional cell carcinoma (BTCC) at different stages and the mechanism by which it inhibits tumor endothelial cell proliferation.MethodsNormal bladder mucosa biopsy specimens were obtained as the control group, and human BTCC biopsy specimens were used as the study group. Immunohistochemical assays were used to detect the expression levels of BAI1, vascular endothelial growth factor (VEGF) and mutant p53, in addition to microvessel density (MVD) in the tissues. Western blotting was used to analyze the differential expression of BAI1 in the two samples.ResultsStatistical analysis was performed, which indicated that BAI1 expression levels in the normal bladder mucosa group were significantly higher than those in the BTCC group and were associated with clinical staging. BAI1 levels in the T1 stage BTCC tissues were higher than those in the T2–4 stage BTCC tissues (P<0.05). BAI1 expression levels were negatively correlated with those of VEGF (r=−0.661, P<0.001), mutant p53 (r=−0.406, P=0.002) and with the MVD (r=−0.675, P<0.001).ConclusionsBAI1 may be involved in the negative regulation of BTCC microvascular proliferation, and its expression may be associated with a reduction in p53 mutations.

Long noncoding RNA HOTAIR is a prognostic biomarker and inhibits chemosensitivity to doxorubicin in bladder transitional cell carcinoma.

In this study, whether HOTAIR is a prognostic biomarker will be detected, and its regulative effects of chemosensitivity to doxorubicin in TCC cells will be examined. The expression of HOTAIR was detected by quantitative real-time PCR. Overall survival rate was calculated by Kaplan-Meier method with the log-rank test for comparisons. MTT assay was used to detect cell proliferation ability and chemosensitivity. Dual-color flow cytometric method was used to detect cell apoptosis. HOTAIR was up-regulated in bladder transitional cell carcinoma (TCC) tissues and cell lines compared with normal bladder transitional cell (NBTC) tissues and bladder epithelial immortalized SV-HUC-1 cells, and its expression level had positive correlation with histological grades of TCC. Moreover, HOTAIR was an independent prognostic biomarker of overall survival for TCC patients. The expression and silence vector for HOTAIR were transfected into T24 and J82 cells to up-regulate and silence the HOTAIR expression, respectively. In T24 and J82 cells, HOTAIR over-expression promoted cell proliferation and inhibited chemosensitivity to doxorubicin and cell apoptosis induced by doxorubicin; silence of HOTAIR showed opposite regulative effects. In summary, lncRNA HOTAIR was an independent prognostic biomarker of overall survival in TCC patients and could regulate chemosensitivity to doxorubicin of human TCC cells. HOTAIR might provide a new potential therapeutic target and stratagem for TCC.

Decreased expression of protein tyrosine phosphatase non-receptor type 12 is involved in the proliferation and recurrence of bladder transitional cell carcinoma.

Protein tyrosine phosphatase non-receptor type 12 (PTPN12) has been shown to be involved in the development of a number of types of carcinoma. However, the effect of PTPN12 on the proliferation and recurrence of human bladder transitional cell carcinoma (TCC) is unclear. The present study aimed to investigate the expression and function of PTPN12 in human TCC. Samples from 164 patients with TCC, in addition to 146 patients undergoing bladder surgery for indications other than TCC, were examined. PTPN12 protein expression was examined using immunohistochemistry and western blotting, and PTPN12 mRNA expression was examined using reverse transcription-quantitative polymerase chain reaction. PTPN12 expression was increased following transfection with the PTPN12-expressing, pcDEF3 vector, and PTPN12 expression was decreased by RNA interference, in four TCC cell lines. The proliferation of TCC cells was analyzed by a WST-1 assay and in xenografts on BALB/C nude mice. The effect of PTPN12 on tumor recurrence was analyzed by adhesion, migration and invasion assays in TCC cell lines. PTPN12 expression was significantly decreased in TCC tissues compared with that in normal urothelium, and the level of PTPN12 expression was negatively correlated with tumor size, pathological grade, clinical stage and tumor recurrence. Furthermore, decreased expression of PTPN12 significantly enhanced the proliferation of TCC cells in vitro and in vivo. TCC cells with lower levels of PTPN12 exhibited greater adhesion, migration and invasion. In conclusion, PTPN12 expression is downregulated in human TCC. Restoring PTPN12 activity may represent a novel therapeutic strategy for this disease.

Expression of brain‑specific angiogenesis inhibitor‑1 and association with p53, microvessel density and vascular endothelial growth factor in the tissue of human bladder transitional cell carcinoma.

The aim of the present study was to investigate the expression levels of brain‑specific angiogenesis inhibitor‑1 (BAI‑1) in bladder transitional cell carcinoma (BTCC) at different stages and the mechanism by which it inhibits tumor endothelial cell proliferation. Normal bladder mucosa biopsy specimens were obtained as the control group, and human BTCC biopsy specimens were used as the study group. Immunohistochemical assays were used to detect the expression levels of BAI‑1, vascular endothelial growth factor (VEGF) and mutant p53, in addition to microvessel density (MVD) in the tissues. Western blotting was used to analyze the differential expression of BAI‑1 in the two samples. Statistical analysis was performed, which indicated that BAI‑1 expression levels in the normal bladder mucosa group were significantly higher than those in the BTCC group and were associated with clinical staging. BAI‑1 levels in the T1 stage BTCC tissues were higher than those in the T2‑4 stage BTCC tissues (P<0.05). BAI‑1 expression levels were negatively correlated with those of VEGF (r=‑0.661, P<0.001), mutant p53 (r=‑0.406, P=0.002) and with the MVD (r=‑0.675, P<0.001). BAI‑1 may be involved in the negative regulation of BTCC microvascular proliferation, and its expression may be associated with a reduction in p53 mutations.

Quantitation of rare circulating tumor cells by folate receptor α ligand-targeted PCR in bladder transitional cell carcinoma and its potential diagnostic significance.

Numerous attempts for detection of circulating tumor cells (CTC) have been made to develop reliable assays for early diagnosis of cancers. In this study, we validated the application of folate receptor α (FRα) as the tumor marker to detect CTC through tumor-specific ligand PCR (LT-PCR) and assessed its utility for diagnosis of bladder transitional cell carcinoma (TCC). Immunohistochemistry for FRα was performed on ten bladder TCC tissues. Enzyme-linked immunosorbent assay (ELISA) for FRα was performed on both urine and serum specimens from bladder TCC patients (n = 64 and n = 20, respectively) and healthy volunteers (n = 20 and n = 23, respectively). Western blot analysis and qRT-PCR were performed to confirm the expression of FRα in bladder TCC cells. CTC values in 3-mL peripheral blood were measured in 57 bladder TCC patients, 48 healthy volunteers, and 15 subjects with benign urologic pathologies by the folate receptor α ligand-targeted PCR. We found that FRα protein was overexpressed in both bladder TCC cells and tissues. The levels of FRα mRNA were also much higher in bladder cancer cell lines 5637 and SW780 than those of leukocyte. Values of FRα were higher in both serum and urine specimens of bladder TCC patients than those of control. CTC values were also higher in 3-mL peripheral blood of bladder TCC patients than those of control (median 26.5 Cu/3 mL vs 14.0 Cu/3 mL). Area under the receiver operating characteristic (ROC) curve for bladder TCC detection was 0.819, 95 % CI (0.738-0.883). At the cutoff value of 15.43 Cu/3 mL, the sensitivity and the specificity for detecting bladder cancer are 82.14 and 61.9 %, respectively. We concluded that quantitation of CTCs through FRα ligand-PCR could be a promising method for noninvasive diagnosis of bladder TCC.

MP22-18 IDENTIFICATION OF NOVEL GENE EXPRESSION MARKERS FOR BLADDER CANCER DIAGNOSTICS

You have accessJournal of UrologyBladder Cancer: Detection & Screening1 Apr 2014MP22-18 IDENTIFICATION OF NOVEL GENE EXPRESSION MARKERS FOR BLADDER CANCER DIAGNOSTICS Boris Alekseev, Nikolay Vorobyev, Petr Shegay, Anastasia Zabolotneva, Igor Rusakov, Anton Buzdin, Nurshat Gaifullin, and Olga Kovalchuk Boris AlekseevBoris Alekseev More articles by this author , Nikolay VorobyevNikolay Vorobyev More articles by this author , Petr ShegayPetr Shegay More articles by this author , Anastasia ZabolotnevaAnastasia Zabolotneva More articles by this author , Igor RusakovIgor Rusakov More articles by this author , Anton BuzdinAnton Buzdin More articles by this author , Nurshat GaifullinNurshat Gaifullin More articles by this author , and Olga KovalchukOlga Kovalchuk More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.866AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Molecular diagnostics may play key role in developing techniques for the early detection of non-muscle-invasive bladder cancer (BC) and is based on the detection of molecular changes that occur on initial stages of oncological disease. The aim of our study was to identify new potential BC gene expression markers and to evaluate its sensitivity in BC patients. METHODS To identify new gene expression BC markers we performed both bioinformatic and experimental research. For the bioinformatic assays we analysed publically available gene expression data in the Gene Expression Omnibus database. Experimental research consisted in characterization of the gene expression profiles in transitional cell carcinoma (TCC) and normal bladder tissues samples using two alternative approaches: suppression subtractive hybridization (SSH) and microarray hybridization. By using these approaches we have been able to identify 13 novel gene expression markers with area-under-the-curve (AUC) values >0.7 that, to our knowledge, were not associated with TCC in previous studies, but demonstrated a strong correlation with TCC in our tests (p<0,05). Among these entries, there were six top genes with the AUC scores varying from 0.85 to 0.92. To further characterize these novel potential TCC molecular markers, we performed a quantitative RT-PCR (qRT-PCR) assay of the six “top” genes on the 15 normal bladder and 30 surgically-resected TCC samples. RESULTS The transcriptional profiles for ten genes (including five “top” genes): BCL2, CCNE1, CDH1, IGFBP3, STAT1, TRIB1, NUSAP1, PRC1, UBE2C and TFDP1 correlated not only between the experimental and GEO database datasets, but also with the data obtained using SSH. However, the remaining “top” gene, KIFC1, although clearly differential according to our experimental microarray data, demonstrated an intact expression level in the SSH test. The qRT-PCR data confirmed upregulation of the genes: KIFC1, TRIB1, NUSAP1, PRC1, UBE2C, TFDP1, AURKA1 and UHRF1 in the TCC samples. These data suggest that our top six potential BC biomarkers demonstrate strong association with TCC, as reflected by the AUC qRT-PCR values - KIFC1 (0,77), TRIB1 (0,85), NUSAP1 (0,89), PRC1 (0,89), UBE2C (0,91), TFDP1 (0,87). The overall calculated AUC values place them among the best nine known TCC gene expression markers investigated in this study. CONCLUSIONS The top six new gene expression markers - KIFC1, TRIB1, NUSAP1, PRC1, UBE2C and TFDP1 showed excellent sensitivity and reproducibility. Overall, we present a list of potential BC genetic markers possessing a high diagnostic value. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e241-e242 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Boris Alekseev More articles by this author Nikolay Vorobyev More articles by this author Petr Shegay More articles by this author Anastasia Zabolotneva More articles by this author Igor Rusakov More articles by this author Anton Buzdin More articles by this author Nurshat Gaifullin More articles by this author Olga Kovalchuk More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...