Association Of MiR-141 Expression With Prognosis Of Patients And Inhibition Of Zoledronic Acid To Pentose Phosphate Pathway In Bladder Cancer

Part I Increased mi R-141 expression is associated with diagnosis and favorable prognosis of patients with bladder cancerObjective : To analyze the association of micro RNA-141(mi R-141) with the clinicopathological parameters of bladder cancer and evaluate the value of mi R141 in predicting the prognosis of bladder cancer.Methods:In this study, 114 patients with bladder cancer were enrolled in the study, and tissue specimens were obtained from the tumor zone and adjacent normal area. Mi R-141 expression was determined using SYRB Green quantitative real-time polymerase chain reaction assay and was further correlated with patients’ clinicopathological parameters and the follow-up data. Comparison of mi R-141 expression between the tumor tissue and adjacent normal bladder tissue was made using paired t-test.The t-test was applied to evaluate the association between mi R-141 expression and tumor grade(WHO 2004) and muscle invasion status, while one-way ANOVA test to evaluate the association between mi R-141 expression and tumor stage. The association between mi R-141 expression and cancerspecific survival was assessed by the log-rank test and Cox proportional hazard regression analysis, and tumor stage, tumor grade, and gender were entered into the multivariate analysis.Results:The results indicated that mi R-141 was upregulated in malignant bladder specimens compared with normal ones(P<0.001). mi R-141 expression was significantly associated with tumor stage(P<0.001), tumor grade(P<0.001), and muscle invasion status (P<0.001). Log-rank test showed that the higher mi R-141 expression was associated with longer disease-specific survival of the patients with bladder cancer(P<0.001), which was also proven by univariate and multivariate Cox regression analysis(P<0.001 and P=0.039, respectively). Focusing on patients with non-muscle invasive bladder cancer,univariate analysis using log-rank test and Cox regression analysis found that patients with high mi R-141 expression experienced longer disease-free survival(P= 0.031 and P=0.040,respectively)and disease-specific survival(P=0.028 and P=0.038, respectively), which was confirmed by multivariate Cox regression analysis(P=0.036 and P=0.042, respectively).Conclusions:This study showed that mi R-141 may contribute to the progression of bladder cancer and its upregulation may be independently associated with favorable prognosis of bladder cancer, suggesting that mi R-141 might serve as a promising biological marker for further risk stratification in the management of bladder cancer.Part Ⅱ Zoledronic acid inhibits the pentose phosphate pathway through attenuating the Ras-TAp73-G6 PD axis in bladder cancer cellsObjective:To investgate the infection of Zoledronic acid(ZA) to cell proliferation and pentose phosphate pathway in bladder cancer cells.Methods:Glucose consumption was measured in the cell lysates with Glucose Uptake Colorimetric Assay kit.The production of NADPH was measured in the cell lysates with NAD+/NADH Quantification Colorimetric kit.To examine the expression of G6 PD and TAp73 in T24 cells, RT-q PCR and western-blot were performed. After p CMV6-TAp73 vector were transfected into cells, to examine the infection of ZA to pentose phosphate pathway. FPT inhibitor II was substituted for ZA,then examine the levels of Ras-GTP,TAp73, G6 PD,glucose consumption and NADPH production. Following transfection with TAp73 for 48 h, the inhibitory effect of G6 PD, glucose consumption and NADPH production was measured.After knockdown of TAp73, examine the levels of G6 PD,glucose consumption and cell proliferation.Results: ZA inhibited cell proliferation and the pentose phosphate pathway(PPP) in bladder cancer cells. In addition, the expression of glucose-6-phosphate dehydrogenase(G6PD) was found to be inhibited by ZA. Furthermore, the stability of TAp73, which activates the expression G6 PD was decreased in zoledronic acid treated cells. Decreased levels of Ras-GTP were also observed following treatment with ZA. The inhibition of Ras activity by PT inhibitor II also significantly reduced the levels of TAp73 and G6 PD and the PPP flux. Following transfection with TAp73 for 48 h, the inhibitory effect on the PPP by ZA was attenuated by TAp73 overexpression.Moreover, knockdown of TAp73, attenuated the PPP flux and eliminated the affection of ZA on the PPP flux.Conclusions:ZA can inhibit stability of TAp73 and attenuate the PPP via blocking Ras signaling in bladder cancer cells.