miR-141与膀胱癌预后相关性及药物对膀胱癌葡萄糖代谢影响

发布时间：2018-03-11 10:54

本文选题：miR-141　切入点：膀胱癌　出处：《苏州大学》2015年博士论文　论文类型：学位论文

【摘要】：第一部分mi R-141与膀胱癌诊断预后相关性研究目的:分析微小RNA-141(mi R-141)与膀胱癌临床病理分期相关性及评估mi R-141预测膀胱癌预后价值。方法:选取114例膀胱癌患者癌肿及癌旁正常组织,采用实时荧光定量PCR方法检测mi R-141并与患者临床病理分期及随访数据进行相关性分析。比较肿瘤组织和癌旁组织中mi R-141表达水平差异,mi R-141表达与肿瘤分级、是否肌层浸润间关系采取t检验。mi R-141表达和肿瘤分期间相关性分析采取单因素方差分析。mi R-141和肿瘤特异性生存间相关性采取log-rank检验和Cox回归分析,同时将肿瘤分期、分级、患者性别进行多变量方差分析。结果:显示膀胱癌组织中mi R-141表达增高,并与肿瘤分级(P0.001)、分期(P0.001)、侵袭性(P0.001)相关。Log-rank检验显示高mi R-141表达和膀胱癌患者较长无病生存期呈相关性(P0.001),同时使用单变量分析和COX回归分析得到证实(P0.001 and P=0.039)。对于非肌层浸润性膀胱癌患者,单因素log-rank和COX分析显示高mi R-141表达和无病生存期相关I(P=0.031,P=0.040),亦和疾病特异性生存相关(P=0.028,P=0.038),同时使用多因素COX分析亦得到证实(P=0.036,P=0.042)。结论:mi R-141与膀胱癌进展相关,其表达增高成为膀胱癌预后良好独立预测指标。表明mi R-141可成为膀胱癌危险分层的潜在生物学指标。第二部分唑来膦酸在膀胱癌葡萄糖代谢中作用机制研究目的:研究唑来膦酸对膀胱癌细胞生长和葡萄糖代谢的影响。方法:比较唑来膦酸干预前后膀胱癌细胞生长、葡萄糖的摄入和NADPH产生的差异,定量RT-PCR结合western blot检测TAp73和葡萄糖-6-磷酸脱氢酶(G6PD)表达差异;通过建立TAp73过表达的膀胱癌细胞系,验证唑来膦酸对磷酸戊糖途径的抑制作用是否在TAp73过表达的情况下有所改变;以Ras抑制剂替代唑来膦酸处理膀胱癌细胞,western blot检测Ras活性,TAp73和G6PD的表达,检测细胞葡萄糖的摄入和NADPH产生的差异;敲除TAp73后,检测G6PD的表达、葡萄糖摄取和细胞增殖水平。结果:唑来膦酸在膀胱癌细胞中能够抑制细胞增殖和磷酸戊糖途径(PPP)、G6PD。此外,唑来膦酸处理后,TAp73稳定性、Ras-GTP表达水平亦出现下降。Ras抑制剂替代唑来膦酸处理膀胱癌细胞后亦能够显著减少TAp73,G6PD和PPP表达水平。唑来膦酸对于磷酸戊糖途径的抑制作用,在TAp73过表达的影响下受到削弱。应用sh RNA干扰TAp73后能够减少PPP表达水平同时消除唑来膦酸对于PPP表达水平的影响。结论:在膀胱癌细胞中,唑来膦酸通过抑制Ras活性,从而阻断TAp73介导的PPP过度激活,达到抗肿瘤效果。
[Abstract]:Part I the correlation between miR-141 and the diagnosis and prognosis of bladder cancer objective: to analyze the correlation between minute RNA-141(mi R-141) and clinicopathological stage of bladder cancer and to evaluate the prognostic value of mi R-141 in bladder cancer. Methods: 114 cases of bladder cancer were selected. And normal tissues adjacent to cancer, Real-time fluorescence quantitative PCR was used to detect the expression of miR-141 and its correlation with clinical pathological stage and follow-up data of patients. The difference of expression level of miR-141 in tumor tissues and paracancerous tissues was compared between the expression of mi R-141 and tumor grade. Whether the relationship between myometrial invasion was analyzed by t test .mi R-141 expression and the correlation between tumor stage and tumor stage. One-factor ANOVA .miR-141 and tumor-specific survival correlation were analyzed by log-rank test and Cox regression analysis, and tumor staging and grading were used. Results: the expression of mi R-141 was increased in bladder cancer tissues. Log-rank test showed that the expression of high mi R-141 was correlated with the longer disease-free survival time of bladder cancer patients, and the univariate analysis and COX regression analysis were used to confirm that P0.001 and P0. 039. Patients with invasive bladder cancer, Univariate log-rank and COX analysis showed that high miR-141 expression was associated with disease-free survival time, and was also associated with disease-specific survival. Multivariate COX analysis was also used to confirm the relationship between the expression of high miR-141 and disease-free survival. Conclusion\% mi R-141 is associated with the progression of bladder cancer. The increased expression of zoledronic acid may be an independent predictor of good prognosis of bladder cancer, which indicates that mi R-141 may be a potential biological marker for the risk stratification of bladder cancer. Part two: study on the Mechanism of Zoledronic Acid in glucose Metabolism in bladder Cancer. To study the effects of zoledronic acid on the growth and glucose metabolism of bladder cancer cells. The difference between glucose intake and NADPH production, quantitative RT-PCR combined with western blot was used to detect the difference between TAp73 and G6PD. the overexpression of TAp73 in bladder cancer cell line was established. To verify whether the inhibitory effect of zoledronic acid on pentose phosphate pathway was changed under the condition of TAp73 overexpression, and to detect the expression of TAp73 and G6PD in bladder cancer cells treated with Ras inhibitor instead of zoledronic acid by western blot. After knockout TAp73, the expression of G6PD, glucose uptake and cell proliferation were detected. Results: Zoledronic acid inhibited cell proliferation and pentose phosphate pathway in bladder cancer cells. The expression of Ras-GTP decreased after zoledronic acid treatment. The expression of TAp73 G6PD and PPP in bladder cancer cells was significantly decreased after treatment with zoledronic acid instead of zoledronic acid. The inhibitory effect of zoledronic acid on pentose phosphate pathway was observed. Using sh RNA to interfere with TAp73 can reduce the expression of PPP and eliminate the effect of zoledronic acid on the expression of PPP. Conclusion: in bladder cancer cells, zoledronic acid inhibits the activity of Ras. In order to block TAp73-mediated excessive activation of PPP to achieve anti-tumor effect.
【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.14

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