本文選題：膀胱癌 + miR-143��； 參考：《南京醫(yī)科大學》2015年博士論文

【摘要】：背景:miR-143是microRNA的一種,已被證實在多種腫瘤組織中低表達,并可以起到抑癌基因的作用,體外細胞實驗中證實IGF-IR3'-UTR區(qū)存在miR-143調控位點,高表達的miR-143能作用于IGF-IR來抑制腫瘤細胞增殖、侵襲、轉移。microRNA在基因調控中所起的作用受到越來越多的重視,有關在膀胱癌細胞中miR-143調控IGF-IR影響吉西他濱化療敏感性的研究尚未見報道。我們的研究提示miR-143可能在膀胱癌細胞的IGF-IR基因調控機制中起到重要的作用,探索miR-143介導的IGF-IR基因調控作用對膀胱癌細胞生物學行為和化療藥物敏感性的影響十分重要。目的:明確miR-143在膀胱癌組織標本和細胞中的表達水平及其與膀胱癌細胞生長的相關性,探討miR-143影響膀胱癌惡性生物學行為和對化療藥物敏感性的分子機制。方法:1、RT-PCR方法檢測23例膀胱癌及其相對應的癌旁正常膀胱組織標本中miR-143的表達水平,并檢測膀胱癌細胞株T24和5637細胞中miR-143的表達水平,分析miR-143表達與膀胱癌分期的相關性。2、RT-PCR方法檢測23例膀胱癌及其相對應的癌旁正常膀胱組織標本{中IGF-IR的表達水平,分析miR-143與IGF-IR表達的相關性。構建miR-143過表達慢病毒系統(tǒng),轉染膀胱癌細胞。RT-PCR方法檢測過表達miR-143的T24和5637細胞中IGF-IR的表達水平。3、CCK-8法檢測過表達miR-143和干擾IGF-IR對5637細胞增殖的影響。同時檢測過表達miR-143和干擾IGF-IR對吉西他濱敏感性影響。4、Western blotting 實驗分析過表達 miR-143 和干擾 IGF-IR 后 p-AKT、AKT、p-ERK和ERK的蛋白表達情況。結果:1、23對膀胱癌組織標本中,miR-143在膀胱癌組織中表達低于癌旁正常膀胱組織,并且其表達水平與腫瘤的分期密切相關,在膀胱癌細胞株T24和5637中miR-143表達降低。2、miR-143和IGF-IR在膀胱癌組織標本中的表達呈負相關,在T24和5637細胞中高表達的miR-143能明顯抑制IGF-IR表達。3、在5637細胞中過表達miR-143和干擾高表達IGF-IR均能抑制細胞的生長。過表達miR-143和干擾高表達IGF-IR的5637細胞對吉西他濱的敏感性增加。4、上調miR-143能抑制5637細胞中P-AKT和P-ERK的表達。干擾5637細胞中IGF-IR表達可以調節(jié)AKT和ERK信號通路的激活來抑制P-AKT和P-ERK的表達。結論:1、miR-143在膀胱癌組織和細胞中廣泛低表達,與膀胱癌的惡性生物學行為密切相關。2、miR-143抑制膀胱癌細胞的增殖能力,并且可以通過靶向IGF-IR影響AKT和ERK信號通路起到抑癌基因的作用。3、miR-143和吉西他濱對膀胱癌細胞生長有協(xié)同抑制作用,miR-143可以增加膀胱癌細胞對化療藥物(吉西他濱)的敏感性。
[Abstract]:Background: miR-143 is a kind of microRNA, which has been proved to be low expressed in many kinds of tumor tissues and can act as a tumor suppressor gene. In vitro, it has been proved that there are miR-143 regulatory sites in the IGF-IR3'-UTR region, and that high expression miR-143 can act on IGF-IR to inhibit the proliferation of tumor cells. More and more attention has been paid to the role of invasion, metastasis, microRNAs in gene regulation. The effect of miR-143 on the chemosensitivity of gemcitabine in bladder cancer cells has not been reported. Our results suggest that miR-143 may play an important role in the regulation of IGF-IR gene in bladder cancer cells. It is important to explore the effects of miR-143 mediated IGF-IR gene regulation on the biological behavior and chemosensitivity of bladder cancer cells. Objective: to investigate the expression of miR-143 in bladder cancer tissues and its correlation with the growth of bladder cancer cells, and to explore the molecular mechanism of miR-143 affecting malignant biological behavior and chemosensitivity of bladder cancer. Methods the expression of miR-143 in 23 cases of bladder cancer and its adjacent normal bladder tissues was detected by using 1: 1 RT-PCR, and the expression of miR-143 in bladder cancer cell lines T24 and 5637 was also detected. To analyze the correlation between the expression of miR-143 and the stage of bladder cancer. 2RT-PCR was used to detect the expression of IGF-IR in 23 cases of bladder cancer and its adjacent normal bladder tissues, and to analyze the correlation between miR-143 and IGF-IR expression. MiR-143 overexpression lentivirus system was constructed. The expression level of IGF-IR in T24 and 5637 cells overexpressed with miR-143 was detected by RT-PCR. The effect of overexpression of miR-143 and interference of IGF-IR on the proliferation of 5637 cells was detected by the method of CCK-8. The effect of overexpression of miR-143 and interference of IGF-IR on the sensitivity of gemcitabine. 4Western blotting assay was used to analyze the expression of p-AKTT-AK p-ERK and ERK after IGF-IR interference and overexpression of miR-143. Results the expression of miR-143 in bladder cancer tissues was lower than that in adjacent normal bladder tissues, and the expression level was closely related to the stage of the tumor. The expression of miR-143 and IGF-IR in bladder cancer cell lines T24 and 5637 were negatively correlated. Overexpression of miR-143 in T24 and 5637 cells significantly inhibited the expression of IGF-IR. 3. Overexpression of miR-143 and interference of overexpression of IGF-IR in 5637 cells inhibited the growth of T24 and 5637 cells. Overexpression of miR-143 and overexpression of IGF-IR increased the sensitivity of 5637 cells to gemcitabine. Upregulation of miR-143 inhibited the expression of P-AKT and P-ERK in 5637 cells. Interfering with the expression of IGF-IR in 5637 cells can inhibit the expression of P-AKT and P-ERK by regulating the activation of AKT and ERK signaling pathway. Conclusion the low expression of 1: 1 miR-143 in bladder cancer tissues and cells is closely related to the malignant biological behavior of bladder cancer. It can inhibit the proliferation of bladder cancer cells. Moreover, targeting IGF-IR could inhibit the growth of bladder cancer cells by targeting AKT and ERK signaling pathway. 3miR-143 and gemcitabine had synergistic inhibitory effects on the growth of bladder cancer cells. MiR-143 could increase the sensitivity of bladder cancer cells to chemotherapeutic drugs (gemcitabine).
【學位授予單位】：南京醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2015
【分類號】：R737.14

【參考文獻】

相關期刊論文 前2條

1 Xiao-Li Wu;Bin Cheng;Pei-Yuan Li;Huan-Jun Huang;Qiu Zhao;Zi-Li Dan;Dean Tian;Peng Zhang;;MicroRNA-143 suppresses gastric cancer cell growth and induces apoptosis by targeting COX-2[J];World Journal of Gastroenterology;2013年43期

2 董勝國,紀祥瑞,侯四川,邵世修,申東亮;影響膀胱癌患者長期生存的因素分析[J];臨床泌尿外科雜志;1999年06期

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本文編號：1905984