Urothelial Cell Carcinoma Of Bladder Research Articles

MiR-99a-5p acts as tumor suppressor via targeting to mTOR and enhances RAD001-induced apoptosis in human urinary bladder urothelial carcinoma cells.

IntroductionmiR-99a-5p, known to play an important role in mammalian target of rapamycin (mTOR) regulation, is downregulated in human bladder cancer. The study aimed to investigate the anticancer activity of miR-99a-5p and the possible mechanism associated with mTOR in bladder cancer cells.Materials and methodsVectors expressing miR-99a-5p were transfected into human urinary bladder urothelial carcinoma (5637 and T24) cells. The level of miR-99a-5p was monitored by microRNA (miRNA) quantitative polymerase chain reaction (QPCR). Luciferase reporter assays were performed to verify the direct binding of miR-99a-5p to mTOR transcripts. The mTOR transcripts and protein levels were measured by QPCR and Western blot, respectively. Cell viability of miR-99a-5p-transfected cells was detected by tetrazolium salt (WST-1). Inhibition of mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2) signaling was detected by the phosphorylation of mTOR and AKT using Western blot. The ability of miR-99a-5p to enhance RAD001-induced apoptosis was determined as the expression of cleaved caspase 3 and levels of DNA fragmentation.ResultsTransfection of miR-99a-5p-expressing vector elevated the expression level of miR-99a-5p up to sixfold compared to vector-only controls. The results from luciferase assay verified that miR-99a-5p directly binds to the predicted sequence in the 3′ untranslated region (3′-UTR) of mTOR. The levels of mTOR RNA and protein were decreased in miR-99a-5p-transfected cells. Dual inhibition of mTORC1 and mTORC2 by miR-99a-5p was confirmed by the decreased phosphorylation of mTOR (at Ser2448 and Ser2481), phospho-rpS6 and phospho-4EBP1. The phosphorylation of AKT was significantly inhibited in miR-99a-5p-transfected cells upon RAD001 treatment. Enforced expression of miR-99a-5p potentiated RAD001-induced apoptosis in these cells.ConclusionThis is the first study showing that miR-99a-5p markedly inhibits the growth of bladder cancer cells via dual inhibition of mTORC1 and mTORC2. Our data demonstrated that forced expression of miR-99a-5p inhibits the feedback of AKT survival pathway and enhances the induction of apoptosis in RAD001-treated bladder cancer cells.

The relation between Ring Box-1 protein overexpression and tumor grade and stage in bladder urothelial cell carcinoma.

Ring Box Protein-1 (RBX-1), a component of SCF E3 ubiquitin ligases, has a crucial role in bladder urothelial cell carcinoma (UCC) carcinogenesis and progression. In the present study, it is aimed to determine the expression of RBX-1 protein in bladder UCC and the association between tumor grade, stage and RBX-1 expression. Ninety UCC samples and 20 samples containing foci of normal bladder urothelium were recruited and analyzed immunohistochemically in terms of RBX-1 expression. Immuno-reactivity scoring system (IRS) was used to determine RBX-1 expression levels. RBX-1 overexpression was associated with high tumor grade (p= 0.001) and advanced stage (p= 0.001). pT1 tumors showed higher RBX-1 expression than pTa tumors. pT2 tumors showed not only higher expression than pTa tumors but also higher expression than the total of pTa and pT1 groups combined. There was no statistically significant relation between RBX-1 expression and patient gender (p= 0.116) or age (p= 0.191). In bladder UCC, RBX-1 overexpression is associated with high tumor grade and advanced stage and represents biological potential of invasiveness and aggressive disease. Results of the present study have to be supported with further studies to reveal clinical and therapeutic implications of RBX-1 overexpression in bladder UCC.

Knockdown of long non-coding RNA Taurine Up-Regulated 1 inhibited doxorubicin resistance of bladder urothelial carcinoma via Wnt/β-catenin pathway.

In genitourinary system, bladder cancer (BC) is the most common and lethal malignant tumor, which most common type is bladder urothelial carcinoma (BUC). Long non-coding RNA (lncRNA) Taurine Up-Regulated 1 (TUG1) gene is high-expressed in several malignant tumors, including BC. In this study, over-expression of TUG1 was found in BUC tissues and cell line resistant to doxorubicin (Dox). Knockdown of TUG1 inhibited the Dox resistance and promoted the cytotoxicity induced by Dox in T24/Dox cells. TUG1 knockdown also depressed the Wnt/β-catenin pathway, and the activation the Wnt/β-catenin pathway partly reversed the inhibitory effects of TUG1 knockdown on Dox resistance in T24/Dox cells. In conclusion, up-regulation of lncRNA TUG1 was related with the poor response of BUC patients to Dox chemotherapy, knockdown of TUG1 inhibited the Dox resistance of BUC cells via Wnt/β-catenin pathway. These findings might assist in the discovery of novel potential diagnostic and therapeutic target for BUC, thereby improve the effects of clinical treatment in patients.

LncRNA plasmacytoma variant translocation 1 is an oncogene in bladder urothelial carcinoma.

Bladder cancer (BC) is the most lethal malignant cancer of the genitourinary system, and bladder urothelial carcinoma (BUC) is the most common type of BC. The long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) is overexpressed in several malignant tumors, including BC. Using a lncRNA array and quantitative real-time PCR, we detected greater expression of PVT1 in BUC tissues and cell lines resistant to doxorubicin (DOX) and cisplatin (DDP) than in DOX- and DDP-sensitive cells. PVT1 knockdown reduced proliferation and invasion by a DOX- and DDP-resistant T24/DR BUC cells, arrested cells in G1 phase, and increased apoptosis. PVT1 knockdown also sensitized T24/DR cells to DOX and DDP, and suppressed expression of multidrug resistance 1 (MDR1) and multidrug resistance associated protein 1 (MRP1). Wnt/β-catenin pathway activation in T24/DR cells reversed the effects of PVT1 knockdown on metastasis-associated behavior and chemoresistance. In sum, lncRNA PVT1 is overexpressed in multidrug resistant BUC tissues and cell lines, and PVT1 knockdown reduces BUC cell proliferation, invasiveness, and chemoresistance by modulating Wnt/β-catenin signaling. These results provide new insight into BUC chemoresistance mechanisms and suggest potential therapeutic targets for anti-BUC therapeutics.

The role of the tumor microenvironment in bladder cancer development and progression

Urothelial cell carcinoma (UCC) of the bladder comprises a mixture of heterogeneous tumor cell populations, the surrounding stroma (which is populated by different types of mesenchymal cells), and the extracellular matrix (ECM). Bladder cancer (BC) has certain characteristics that distinguish it from other cancers. First, BCs are categorized into two groups: non-muscle invasive BC (NMIBC) and muscle invasive BC (MIBC). The overall survival rate of patients with NMIBC is excellent compared with that of patients with other malignancies; however, some patients have a high risk of recurrence and a variable risk of progression despite administration of local therapies. The oft-cited “50% overall survival at 5 years” for MIBC has remained relatively unchanged over the last 20 years. Second, BC is a highly immunogenic cancer that shows a high rate of mutation due to the fact that more mutations are associated with a higher chance of tumor antigens triggering an appropriate immune response. The host immune response to tumor cells is based on the interactions that take place within the tumor microenvironment (TME). Intravesical bacillus Calmette-Guerin (BCG) therapy is the first U.S. Food and Drug Administration (FDA)-approved immunotherapy for BC and is the most successful immunotherapy for any established human neoplasm. Recently, the FDA approved the use of atezolizumab (Tecentriq ® ) and nivolumab (Opdivo ® ), which mimic programmed cell death ligand-1 inhibitor, to treat patients with locally advanced or metastatic UCC. Combination therapies involving cytotoxic chemotherapy, antiangiogenic agents, alternative immune checkpoint inhibitors, immunostimulatory cytokines, and cancer vaccines are currently under clinical investigation. This review summarizes recent studies of tumor-stromal crosstalk during pathogenesis of UCC of the bladder, and discusses the emerging roles of functionally important cellular components that interact within the cancer cell-TME. In addition, TME-targeted anticancer strategies such as chemotherapy, chemo-immunotherapy, and immunotherapy are briefly reviewed.

Diagnostic value of DNA aneuploidy for urothelial carcinoma.

e16007 Background: The incidence and mortality of urologic cancer are increasing in China and worldwide. Early diagnosis is the key to improve survival. DNA-Image-Cytometry (DNA-ICM) is to assist in the early microscopical diagnosis and grading of various types of tumors, measuring the DNA-contents of their cells. This study investigates the role of DNA aneuploidy in improving diagnostic sensitivity and specificity for urothelial carcinoma. Methods: 274 voided urine samples (204 cases from patients with urothelial carcinomas and 70 cases from patients with benign urinary system diseases) were analyzed with DNA-ICM and liquid-based cytology (LBC) .Fresh urine samples were collected in the morning of the analysis. Sedimented cell-preparations were obtained through centrifugation (cytospins) for DNA-ICM. The sensitivity and specificity of DNA ICM and LBC were calculated by comparing with the pathology in cells with moderate and severe dysplasia. Results: Sensitivity of DNA-ICM was 70.3% which was much higher than that of liquid-based cytology (27.2%)(P < 0.01). Specificity of DNA-ICM was 100% while specificity of LBC was 98.6%. The sensitivity of DNA-ICM in upper urinary tract urothelial cell carcinoma(UTCUs) were 68.75%, lower than 75.00% in bladder urothelial cell carcinomas, there was no significant differences between them(P > 0.01). Conclusions: DNA single cell cytometry represents a highly relevant tool in the objective identification of urothelial carcinoma with a sufficient sensitivity and specificity. Enhancing its extended use as an adjuvant strategy for the early screening of the urothelial carcinoma during routine analyses is benefical. DNA-ICM is an effective screening method for urothelial cancer. Its use to monitor the treatment should be further validated in larger series.

The adenosine A2b receptor promotes tumor progression of bladder urothelial carcinoma by enhancing MAPK signaling pathway.

The adenosine A2b receptor (A2bR) was considered to play an oncogenic role in many human malignancies. However, the expression and molecular function of A2bR in bladder urothelial carcinoma (BUC) have not been well elucidated. Herein, we found that the expression of A2bR was higher than other adenosine receptors in BUC tissues and cells, and it was upregulated in BUC tissues compared with matched normal bladder tissues. Furthermore, high expression of A2bR was associated with poor prognosis of patients with BUC. In addition, suppression of A2bR inhibited the proliferation, migration and invasion of BUC cells and arrested the cell cycle at the G1 phase. Finally, we demonstrated that downregulation of A2bR inhibited the proliferation, migration and invasion of BUC in part via the MAPK signaling pathway, increasing the levels of P21 but decreasing the levels of cyclin B1, D, E1, MMP-2 and MMP-9. Overexpression of MMP-2 could rescue BUC cells migration and invasion. Thus, the present study indicates that A2bR may play a potential oncogenic role in BUC progression and act as a potential biomarker to identify BUC patients with poor clinical outcomes.

MP44-01 IN VIVO FLUORESCENCE IMAGING OF AN ORTHOTOPIC RAT BLADDER TUMOR MODEL INDICATES DIFFERENTIAL UPTAKE OF INTRAVESICALLY INSTILLED NEAR-INFRARED LABELED 2-DEOXYGLUCOSE ANALOG BY NEOPLASTIC URINARY BLADDER TISSUES

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology I1 Apr 2017MP44-01 IN VIVO FLUORESCENCE IMAGING OF AN ORTHOTOPIC RAT BLADDER TUMOR MODEL INDICATES DIFFERENTIAL UPTAKE OF INTRAVESICALLY INSTILLED NEAR-INFRARED LABELED 2-DEOXYGLUCOSE ANALOG BY NEOPLASTIC URINARY BLADDER TISSUES Joel Slaton, Carole Davis, Robert Hurst, and Daqing Piao Joel SlatonJoel Slaton More articles by this author , Carole DavisCarole Davis More articles by this author , Robert HurstRobert Hurst More articles by this author , and Daqing PiaoDaqing Piao More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.1332AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Bladder cancer is one of the most expensive cancers to manage due to frequent recurrences requiring life-long surveillance and treatment. Deoxyglucose radionucleotides have been shown to detect metastatic bladder cancer. This pilot study has explored differential uptake of intravesically administered IRDye800CW-DG, a near-infrared labeled 2-deoxy-d-glucose probe targeting glucose metabolism pathway, to detect surface cancer in an orthotopic rodent bladder tumor model. METHODS Twenty-five female Fischer rats were randomly grouped to four conditions: control (n=3), control and intravesically instilled with IRDye800CW-DG (n=6), treated with GFP-labeled AY-27 rat bladder urothelial cell carcinoma cells and washed with saline (n=5), and treated with AY-27 and intravesically instilled with IRDye800CW-DG (n=11). Near-infrared fluorescence was measured from the opened bladder wall of anesthetized rat at an excitation wavelength of 750nm and an emission wavelength of 776nm, by using an in-house fluorescence imaging system. RESULTS There is no statistically significant difference of the peak fluorescence intensity among the control bladders (n=3), the control bladders instilled with IRDye800CW-DG (n=6), and the GFP-labeled AY-27 treated bladders washed by saline (n=5). When compared to that of the control bladders instilled with IRDye800CW-DG (n=6), the fluorescence intensity of GFP-labeled AY-27 treated bladders instilled with IRDye800CW-DG and with histology confirmed neoplastic bladder tissue (n=11) was remarkably stronger (3.34 folds of the former) that was also statistically significant (p<0.0001). CONCLUSIONS The differential uptake of IRDye800CW-DG by the neoplastic urinary bladder tissues suggests the potential for cystoscopy-adaptation to enhance diagnosis and guiding surgical management of flat urinary bladder cancer. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e563 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Joel Slaton More articles by this author Carole Davis More articles by this author Robert Hurst More articles by this author Daqing Piao More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Solute carrier family 12 member 5 promotes tumor invasion/metastasis of bladder urothelial carcinoma by enhancing NF-\u03baB/MMP-7 signaling pathway

Solute carrier family 12 member 5 (SLC12A5), an integral membrane KCl cotransporter, which maintains chloride homeostasis in neurons, is aberrantly expressed and involved in the tumorigenesis of certain cancers. However, the clinical significance and biological role of SLC12A5 in human bladder urothelial carcinoma (BUC) remains unclear. In this study, the expression of SLC12A5 was examined in clinical specimens of primary BUC and in BUC cell lines using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry (IHC). The prognostic value of SLC12A5 expression and its correlation with the clinicopathological features of patients with BUC were analyzed statistically. A series of in vitro and in vivo assays were performed to elucidate the effect of SLC12A5 in BUC and its underlying mechanisms. The present results showed that SLC12A5 expression was significantly increased in BUC tissues. SLC12A5 expression significantly correlated with the tumor node metastasis (TNM) stage. Kaplan–Meier survival curves showed that high SLC12A5 expression was associated with poor survival in patients with BUC. Multivariate analysis indicated that SLC12A5 expression was an independent prognostic marker for the survival of patients. Downregulation of SLC12A5 inhibited the migratory and invasive abilities of BUC cells in vitro, and knocking down SLC12A5 diminished BUC metastasis in vivo. Moreover, we identified that SLC12A5 promoted the migration and invasion of BUC by enhancing MMP-7 expression via NF-κB-dependent transcription. Taken together, our findings indicated that SLC12A5 might function as a tumor metastasis promoting factor in the development and progression of BUC by regulating the NF-κB/MMP-7 signaling pathway. Thus, SLC12A5 might be a prognostic marker as well as a therapeutic target for BUC.

Cytotoxic and toxicogenomic effects of silibinin in bladder cancer cells with different TP53 status.

Silibinin is a natural phenol found in the seeds of the milk thistle plant. Recent data have shown its effectiveness for preventing/treating bladder tumours. Therefore, in this study we investigated the cytotoxic and toxicogenetic activity of silibinin in bladder cancer cells with different TP53 statuses. Two bladder urothelial carcinoma cell lines were used: RT4 (wild-type TP53 gene) and T24 (mutated TP53 gene). Cell proliferation, clonogenic survival, apoptosis rates, genotoxicity and relative expression profile of FRAP/mTOR, FGFR3, AKT2 and DNMT1 genes and of miR100 and miR203 were evaluated. Silibinin promoted decreased proliferation and increased late apoptosis in TP53 mutated cells. Increased early apoptosis rates, primary DNA damage, and decrease of cell colonies in the clonogenic survival assay were detected in both RT4 and T24 cell lines. Down-regulation of FRAP/mTOR, AKT2, FGFR3, DNMT1 and miR100 expression occurred in RT4 cells. Modulation of miR203 was observed in both cell lines. In conclusion, despite the reduction of clone formation in both cell lines, the toxicogenomic effect of silibinin on FRAP/mTOR, AKT2, FGFR3, DNMT1 and miR100 was dependent on the TP53 status. Taken together, the data confirmed the role of silibinin as an antiproliferative compound, whose mechanism of action was related to the TP53 status.

Downregulation of feline sarcoma-related protein inhibits cell migration, invasion and epithelial-mesenchymal transition via the ERK/AP-1 pathway in bladder urothelial cell carcinoma.

Feline sarcoma-related protein (Fer) is a nuclear and cytoplasmic non-receptor protein tyrosine kinase and Fer overexpression is associated with various biological processes. However, the clinicopathological characteristics and molecular mechanisms of Fer expression in bladder urothelial cell carcinoma (UCC) have yet to be elucidated. The present study demonstrated that Fer was significantly upregulated in bladder UCC tissues and cell lines. A clinicopathological analysis suggested that Fer expression was significantly associated with tumor stage, histological grade and lymph node status, and Fer expression was a prognostic factor for overall survival in a multivariate analysis. Furthermore, small interfering RNA (siRNA) was used to silence the expression of the Fer gene in human bladder UCC T24 cells, and was shown to significantly reduce the migration and invasion of the cells. It was also observed that Fer-siRNA caused the T24 cells to acquire an epithelial cobblestone phenotype, and was able to reverse the epithelial-mesenchymal transition of the cells. Subsequently, Fer-knockdown was shown to deactivate the extracellular signal-regulated kinase/activator protein-1 signaling pathway in T24 cells. These results indicated, for the first time, that Fer has a critical role in bladder UCC progression and may be a potential therapeutic target for bladder UCC metastasis.

Evans blue-mediated white-light detection of non-muscle-invasive bladder cancer: A preclinical feasibility and safety study using a rat bladder urothelial cell carcinoma model.

Photodynamic diagnosis (PDD) improves the detection of non-muscle-invasive bladder cancer (NMIBC). However, white-light (WL) cystoscopy remains the technique routinely used in urological clinics. A more cost-effective but equally performant alternative to PDD may encompass the use of an intense tumoritropic dye in combination with WL cystoscopy. Using a preclinical setting, we investigated the practical aspects of the use of Evans blue (EB) dye for the possible future detection of NMIBC using WL cystoscopy. A solution of 1 and 5 mM EB was instilled into healthy and AY-27 tumor-bearing rat bladders. The bladders were then rapidly dissected and the inner walls were inspected for EB using WL stereomicroscopy. EB present in the bladders and the plasma was also quantified using high performance liquid chromatography. To assess the effects of repeated instillations on normal rat bladders, EB was instilled for 7 consecutive days, after which time the bladder wall was investigated histologically. To gain insight into the mechanisms underlying the selective accumulation of EB in malignant urothelium, RNA sequencing of urothelial tissue and subsequent comparative analysis were performed, with a specific focus on cell adhesion. The concentrations of EB were substantially higher in malignant bladders compared with those in healthy bladders, matching the blue staining of the inner bladder wall observed by stereomicroscopy. EB was equally present in the plasma of healthy and tumor-bearing subjects, although at low concentrations. Importantly, EB did not cause any abnormalities in the urothelium after 7 days of repeated instillation in normal rats. RNA sequencing of the urothelium indicated an abnormal expression of several genes related to cell adhesion in malignant urothelium compared with the normal urothelium. Our findings may be important for future clinical developments in the field of diagnostics for bladder cancer. Implementing the more cost-effective protocol of EB instillations in combination with WL cystoscopy may offer a benefit to patients as well as the healthcare system.

Protein Profiling of Bladder Urothelial Cell Carcinoma.

This study aimed to detect protein changes that can assist to understand the underlying biology of bladder cancer. The data showed forty five proteins were found to be differentially expressed comparing tumors vs non-tumor tissues, of which EGFR and cdc2p34 were correlated with muscle invasion and histological grade. Ten proteins (ß-catenin, HSP70, autotaxin, Notch4, PSTPIP1, DPYD, ODC, cyclinB1, calretinin and EPO) were able to classify muscle invasive BCa (MIBC) into 2 distinct groups, with group 2 associated with poorer survival. Finally, 3 proteins (P2X7, cdc25B and TFIIH p89) were independent factors for favorable overall survival.

Role of galectin-1 in urinary bladder urothelial carcinoma cell invasion through the JNK pathway.

Human galectin‐1 is a member of the galectin family, proteins with conserved carbohydrate‐recognition domains that bind galactoside. Galectin‐1 is highly expressed in various tumors and participates in various oncogenic processes. However, detailed descriptions of the function of galectin‐1 in urinary bladder urothelial carcinoma have not been reported. Our previous cohort investigation showed that galectin‐1 is associated with tumor invasiveness and is a possible independent prognostic marker of urinary bladder urothelial carcinoma. The present study aimed to clarify the relevance of galectin‐1 expression level to tumor progression and invasion. In order to decipher a mechanism for the contribution of galectin‐1 to the malignant behavior of urinary bladder urothelial carcinoma, two bladder cancer cell lines (T24 and J82) were established with knockdown of galectin‐1 expression by shRNA. Bladder cancer cells with LGALS1 gene silencing showed reduced cell proliferation, lower invasive capability, and lower clonogenicity. Extensive signaling pathway studies indicated that galectin‐1 participated in bladder cancer cell invasion by mediating the activity of MMP9 through the Ras–Rac1–MEKK4–JNK–AP1 signaling pathway. Our functional analyses of galectin‐1 in urinary bladder urothelial carcinoma provided novel insights into the critical role of galectin‐1 in tumor progression and invasion. These results revealed that silencing the galectin‐1‐mediated MAPK signaling pathway presented a novel strategy for bladder cancer therapy.

The association and prognostic significance of B7-H3 with CD68+ tumor-associated macrophages in urothelial cell carcinoma

Objective To investigate the expression of B7-H3 and the infiltration by CD68+ tumor-associated macrophages (TAMs) in urothelial cell carcinoma (UCC) of the bladder and evaluate its potential prognostic significance for bladder cancer. Methods Data from 76 consecutive UCC patients (63 males and 13 females), median age 65.5 years(ranging from 41 to 86 years), who underwent radical cystectomy, was collected. Immunohistochemistry for B7-H3 expression and CD68+ TAMs infiltration was performed on tumor tissues to explore their association with clinicopathologic features and postoperative survival. Results B7-H3 was widely expressed in UCC, as 54 of 76 (71.1%) tumors showed high expression of positive staining, and significantly increased compared with adjacent, nontumor urothelium (P<0.05). The B7-H3 expression and infiltration of CD68+ TAMs were independent of gender, age, tumor stage, grade, recurrence, lymphoid metastasis, yet both of them were markedly associated with distant metastasis of tumor(P=0.023; P=0.015). In addition, CD68+ TAMs infiltration and B7-H3 expression have significantly positive correlation (P=0.007). Furthermore, Kaplan-Meier analysis showed that overall survival and progression-free survival of patients with high B7-H3 expression was significantly worse than those with low B7-H3 expression (50 months vs. 70 months, P=0.004; 33 months vs. 59 months, P=0.006), while CD68+ TAMs infiltration was not independently associated with OS and PFS in the same cohort. But, in low CD68+ TAMs infiltration subgroup, the median OS and PFS in patients with high B7-H3 expression were shorter than patients with low B7-H3 expression (50 months vs. 68 months, P=0.034; 26 months vs. 59 months, P=0.024). Conclusions This study is the first report of B7-H3 expression and CD68+ TAMs infiltration in urothelial cell carcinoma of bladder in China. B7-H3 is highly expressed in UCC tissues, which could be used alone or in combination with CD68+ TAMs for assessing tumor progression after radical cystectomy. Therefor, these data indicate potentially important prognostic and therapeutic implications of B7-H3 and CD68+ TAMs for patients with UCC. Key words: B7-H3 genes; CD68 genes; Tumor-associated macrophages; Urothelial cell carcinoma; Survival analysis

Overexpression of maelstrom promotes bladder urothelial carcinoma cell aggressiveness by epigenetically downregulating MTSS1 through DNMT3B.

We have recently identified and characterized a novel oncogene, maelstrom (MAEL) from 1q24, in the pathogenesis of hepatocellular carcinoma. In this study, MAEL was investigated for its oncogenic role in urothelial carcinoma of the bladder (UCB) tumorigenesis/aggressiveness and underlying molecular mechanisms. Here, we report that overexpression of MAEL in UCB is important in the acquisition of an aggressive and/or poor prognostic phenotype. In UCB cell lines, knockdown of MAEL by short hairpin RNA is sufficient to inhibit cell growth, invasiveness/metastasis and suppressed epithelial-mesenchymal transition (EMT), whereas ectopic overexpression of MAEL promoted cell growth, invasive and/or metastatic capacity and enhanced EMT both in vitro and in vivo. We further demonstrate that MAEL could induce UCB cell EMT by downregulating a critical downstream target, the metastasis suppressor 1 (MTSS1) gene, ultimately leading to an increased invasiveness of cancer cells. Notably, overexpression of MAEL in UCB cells substantially enhanced the enrichment of DNA methyltrans-ferase (DNMT)3B and histone deacetylase (HDAC)1/2 on the promoter of the MTSS1, and thereby epigenetically suppressing the MTSS1 transcription. Downregulation of MTSS1 by MAEL in UCB cells is partially dependent on DNMT3B. Furthermore, we identify that beside the gene amplification of MAEL, miR-186 is a key negative regulator of MAEL and downregulation of miR-186 is another important mechanism for MAEL overexpression in UCBs. These data suggest that overexpression of MAEL, caused by gene amplification and/or decreased miR-186, has a critical oncogenic role in UCB pathogenesis by downregulation of MTSS1, and MAEL could be used as a novel prognostic marker and/or effective therapeutic target for human UCB.

Outcomes and prognostic factors of simple partial cystectomy for localized bladder urothelial cell carcinoma

Radical cystectomy has remained the gold standard for recurrent superficial or muscle invasive bladder tumor. However, partial cystectomy still has a role in those who reject or have contraindications for radical cystectomy. In this study, we sought to identify predictors of bladder recurrence and overall survival after simple partial cystectomy. We included 27 patients with bladder tumor who received simple partial cystectomy without pelvic lymph node dissection between March 2000 and September 2013. Adjuvant chemotherapy or radiation therapy was prescribed according to the pathological results. Parameters were compared on the basis of bladder recurrence and overall survival. During a mean follow-up time of 39 months, five patients (18.5%) experienced bladder recurrence. An older age, a higher pathological stage, positive surgical margins, and distant metastases were significant predictors of overall survival (p = 0.031, p = 0.001, p = 0.001, and p = 0.011, respectively). Meanwhile, previous bladder instillation and positive surgical margins were significant predictors of bladder recurrence (p = 0.026 and p = 0.027, respectively). The rate of consecutive distant metastases (33.3%) was almost twice the rate of bladder recurrence (18.5%), and six patients developed consecutive distant metastases without first experiencing bladder recurrence. In patients who received a simple partial cystectomy as an alternative treatment, previous bladder instillation and positive surgical margins were significant predictors of bladder recurrence. Patients with an older age, positive surgical margins, and consecutive distant metastases had worse overall survival. Partial cystectomy with routine lymph node dissection may be a better option for achieving favorable long-term outcomes.

Clarification of the molecular pathway of Taiwan local pomegranate fruit juice underlying the inhibition of urinary bladder urothelial carcinoma cell by proteomics strategy.

BackgroundPomegranate fruit has been shown to exhibit the inhibitory activity against prostate cancer and lung cancer in vitro and in vivo, which might be a resource for chemoprevention and chemotherapy of cancer. Our previous documented findings indicated that treatment of urinary bladder urothelial carcinoma cell with the ethanol extract isolated from the juice of pomegranate fruit grown in Taiwan could inhibit tumor cell. In this study we intended to uncover the molecular pathway underlying anti-cancer efficacy of Taiwan pomegranate fruit juice against urinary bladder urothelial carcinoma.MethodsWe exploited two-dimensional gel electrophoresis coupled with tandem mass spectrometry to find the de-regulated proteins. Western immunoblotting was used to confirm the results collected from proteomics study.ResultsComparative proteomics indicated that 20 proteins were differentially expressed in ethanol extract-treated T24 cells with 19 up-regulated and 1 down-regulated proteins. These de-regulated proteins were involved in apoptosis, cytoskeleton regulation, cell proliferation, proteasome activity and aerobic glycolysis. Further studies on signaling pathway demonstrated that ethanol extract treatment might inhibit urinary bladder urothelial carcinoma cell proliferation through restriction of PTEN/AKT/mTORC1 pathway via profilin 1 up-regulation. It also might evoke cell apoptosis through Diablo over-expression.ConclusionsThe results of this study provide a global picture to further investigate the anticancer molecular mechanism of pomegranate fruit.Electronic supplementary materialThe online version of this article (doi:10.1186/s12906-016-1071-7) contains supplementary material, which is available to authorized users.

Expression profile of hydrogen sulfide and its synthases correlates with tumor stage and grade in urothelial cell carcinoma of bladder

BackgroundHydrogen sulfide (H2S) is a newly discovered gas transmitter. It is synthesized by cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (MPST). Endogenous hydrogen sulfide has never been studied in bladder cancer. PurposeWe evaluated H2S production and its synthases expression levels in transitional cell carcinoma (urothelial cell carcinoma of bladder [UCB]) of human bladder tissue and cell lines. Materials and methodsImmunostaining was performed in urothelial cell lines and bladder specimens from 94 patients with UCB of different stages/grades. The expression levels/activities of CBS, CSE, and MPST of specimens and cell lines were analyzed by image semiquantity assay, western blot, and a sulfur-sensitive electrode. We tried to find the correlation between hydrogen sulfide and its synthases with tumor stage in UCB. All experiments were repeated at least 3 times. ResultsImmunoreactivity for CBS, CSE, and MPST was detected in malignant uroepithelium and muscular layer of all tissues examined and cultured cells. The expression levels of CBS, CSE, and MPST were associated with UCB stage/grade. Muscle-invasive bladder cancer samples showed the highest production of H2S (52.6±2.91 nmol/[mg·min]) among all tested samples and EJ cells (transitional cell carcinoma, grade IIIshowed the highest production of H2S among all tested cell lines (53.3±7.02nmol/[mg·min]). ConclusionsProtein levels and catalytic activities of CBS, CSE, and MPST increased with the increase of malignant degrees in human bladder tissues and human UCB cell lines. Our findings may promote the application of these novel enzymes to UCB diagnosis or treatment.

Review of current optical diagnostic techniques for non-muscle-invasive bladder cancer.

IntroductionUrinary bladder urothelial cell carcinoma is one of the most commonly diagnosed cancers in Europe. After prostate, lung and colon cancers, bladder cancer rates as the fourth most common cancer in men in the world.Urinary bladder cancer detection, treatment, and staging have traditionally been based on an endoscopic examination – cystoscopy.Material and methodsA Medline, and Web of Science database search was performed on September 2015 without setting time limits, using the terms ‘bladder cancer’ in conjunction with ‘cystoscopy’, ‘diagnosis’, ‘detection’, ‘fluorescence’, ‘blue-light’, ‘PDD’, ‘narrow band imaging’, ‘molecular imaging’, ‘optical coherence tomography’ or ‘confocal laser endomicroscopy’.ResultsThe new imaging techniques can be classified according to their scope as macroscopic, microscopic, and molecular. Macroscopic techniques, such as narrow band imaging, are similar to white light cystoscopy; however, they help visualize even very minute lesions in the bladder mucosa by means of contrast enhancement.Microscopic imaging techniques, such as optical coherence tomography and confocal laser endomicroscopy, provide high-resolution cross-sectional views of vesicular tissues, which resemble images obtained by histopathological examination. Therefore, these are referred as ‘optical biopsy’. Molecular imaging methods offer highly specific real-time visualization of cancer cells and their differentiation from healthy tissue, by combining optical imaging with fluorescent labeling of elements such as antibodies.ConclusionsIn this article we present a review of studies and literature concerning modern optical diagnostic techniques for non-muscle-invasive bladder cancer. We present available technology with its advantages and disadvantages, and studies regarding its effectiveness.