本文選題：miR-141　切入點：膀胱癌　出處：《蘇州大學(xué)》2015年博士論文　論文類型：學(xué)位論文

【摘要】：第一部分mi R-141與膀胱癌診斷預(yù)后相關(guān)性研究目的:分析微小RNA-141(mi R-141)與膀胱癌臨床病理分期相關(guān)性及評估m(xù)i R-141預(yù)測膀胱癌預(yù)后價值。方法:選取114例膀胱癌患者癌腫及癌旁正常組織,采用實時熒光定量PCR方法檢測mi R-141并與患者臨床病理分期及隨訪數(shù)據(jù)進行相關(guān)性分析。比較腫瘤組織和癌旁組織中mi R-141表達水平差異,mi R-141表達與腫瘤分級、是否肌層浸潤間關(guān)系采取t檢驗。mi R-141表達和腫瘤分期間相關(guān)性分析采取單因素方差分析。mi R-141和腫瘤特異性生存間相關(guān)性采取log-rank檢驗和Cox回歸分析,同時將腫瘤分期、分級、患者性別進行多變量方差分析。結(jié)果:顯示膀胱癌組織中mi R-141表達增高,并與腫瘤分級(P0.001)、分期(P0.001)、侵襲性(P0.001)相關(guān)。Log-rank檢驗顯示高mi R-141表達和膀胱癌患者較長無病生存期呈相關(guān)性(P0.001),同時使用單變量分析和COX回歸分析得到證實(P0.001 and P=0.039)。對于非肌層浸潤性膀胱癌患者,單因素log-rank和COX分析顯示高mi R-141表達和無病生存期相關(guān)I(P=0.031,P=0.040),亦和疾病特異性生存相關(guān)(P=0.028,P=0.038),同時使用多因素COX分析亦得到證實(P=0.036,P=0.042)。結(jié)論:mi R-141與膀胱癌進展相關(guān),其表達增高成為膀胱癌預(yù)后良好獨立預(yù)測指標。表明mi R-141可成為膀胱癌危險分層的潛在生物學(xué)指標。第二部分唑來膦酸在膀胱癌葡萄糖代謝中作用機制研究目的:研究唑來膦酸對膀胱癌細胞生長和葡萄糖代謝的影響。方法:比較唑來膦酸干預(yù)前后膀胱癌細胞生長、葡萄糖的攝入和NADPH產(chǎn)生的差異,定量RT-PCR結(jié)合western blot檢測TAp73和葡萄糖-6-磷酸脫氫酶(G6PD)表達差異;通過建立TAp73過表達的膀胱癌細胞系,驗證唑來膦酸對磷酸戊糖途徑的抑制作用是否在TAp73過表達的情況下有所改變;以Ras抑制劑替代唑來膦酸處理膀胱癌細胞,western blot檢測Ras活性,TAp73和G6PD的表達,檢測細胞葡萄糖的攝入和NADPH產(chǎn)生的差異;敲除TAp73后,檢測G6PD的表達、葡萄糖攝取和細胞增殖水平。結(jié)果:唑來膦酸在膀胱癌細胞中能夠抑制細胞增殖和磷酸戊糖途徑(PPP)、G6PD。此外,唑來膦酸處理后,TAp73穩(wěn)定性、Ras-GTP表達水平亦出現(xiàn)下降。Ras抑制劑替代唑來膦酸處理膀胱癌細胞后亦能夠顯著減少TAp73,G6PD和PPP表達水平。唑來膦酸對于磷酸戊糖途徑的抑制作用,在TAp73過表達的影響下受到削弱。應(yīng)用sh RNA干擾TAp73后能夠減少PPP表達水平同時消除唑來膦酸對于PPP表達水平的影響。結(jié)論:在膀胱癌細胞中,唑來膦酸通過抑制Ras活性,從而阻斷TAp73介導(dǎo)的PPP過度激活,達到抗腫瘤效果。
[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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R737.14

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相關(guān)期刊論文 前1條

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本文編號：1597858