本文選題：肝腫瘤　切入點：慢病毒　出處：《吉林大學(xué)學(xué)報(醫(yī)學(xué)版)》2017年02期 　論文類型：期刊論文

【摘要】：目的:探討肝癌SMMC7721細(xì)胞株P(guān)27RF-Rho mRNA基因沉默對5-氟脲嘧啶(5-Fu)藥物敏感性的影響,為臨床晚期肝癌治療提供理論依據(jù)。方法:構(gòu)建P27RF-Rho RNAi載體,P27RF-Rho基因沉默慢病毒感染SMMC7721肝癌細(xì)胞,Western blotting法檢測基因沉默效果。SMMC7721細(xì)胞分為Scramble-siRNA陰性對照組、5-Fu組、P27RF-Rho-siRNA組和P27RF-Rho-siRNA+5-Fu組。熒光顯微鏡檢測細(xì)胞轉(zhuǎn)染效果,Western blottting法檢測RNAi基因沉默效率,MTT法檢測各組細(xì)胞生長曲線,劃痕實驗檢測細(xì)胞遷移能力,Transwell小室檢測細(xì)胞侵襲能力,Western blotting法檢測各組細(xì)胞中腫瘤相關(guān)蛋白P27及RhoC表達(dá)。結(jié)果:成功構(gòu)建P27RF-Rho RNAi慢病毒載體。Western blotting法,P27RF-Rho-siRNA組細(xì)胞中P27RF-Rho蛋白表達(dá)水平明顯低于5-Fu組和Scramble-siRNA組(P0.05)。與其他3組比較,P27RF-Rho-siRNA+5-Fu組細(xì)胞生長速度降低(P0.05);劃痕實驗中P27RF-Rho-siRNA+5-Fu組細(xì)胞遷移能力明顯減低(P0.01);P27RF-Rho-siRNA+5-Fu組穿過Transwell小室微孔濾膜的平均細(xì)胞數(shù)明顯少于其他3組(P0.01);Western blotting法檢測,P27RF-Rho-siRNA+5-Fu組細(xì)胞中癌相關(guān)蛋白P27表達(dá)水平明顯高于其他3組(P0.05),侵襲相關(guān)蛋白RhoC表達(dá)水平低于其他3組(P0.05)。結(jié)論:P27RF-Rho基因沉默能明顯增強5-Fu對肝癌SMMC7721細(xì)胞的藥物敏感性。
[Abstract]:Objective: To investigate the hepatocellular carcinoma cell line SMMC7721 P27RF-Rho mRNA gene silencing on 5- fluorouracil (5-Fu) effect of drug sensitivity, and provide a theoretical basis for clinical treatment of advanced hepatocellular carcinoma. Methods: to construct P27RF-Rho RNAi vector, P27RF-Rho gene silencing lentivirus infected SMMC7721 cell, detect the gene silencing effect of.SMMC7721 cells Western blotting were divided into Scramble-siRNA negative the control group, 5-Fu group, P27RF-Rho-siRNA group and P27RF-Rho-siRNA+5-Fu group. Fluorescence microscope was used to detect the transfection effect, detection of RNAi gene silencing efficiency of Western blottting method, MTT method to detect the cells growth curve, cell migration and wound healing assay, invasion assay of Transwell cells, the expression of tumor cells were detected by blotting Western and RhoC P27 related protein results: to construct P27RF-Rho RNAi lentivirus vector.Western blotting method, P27RF-Rh The level was significantly lower than that of 5-Fu group and Scramble-siRNA group, the expression of P27RF-Rho protein in o-siRNA cells (P0.05). Compared with other 3 groups, P27RF-Rho-siRNA+5-Fu group cell growth rate decreased (P0.05); group P27RF-Rho-siRNA+5-Fu cell migration in a scratch assay was significantly decreased (P0.01); the average number of cells in the P27RF-Rho-siRNA+5-Fu group through the Transwell chamber of microporous membrane was significantly less than the other 3 group (P0.01); Western blotting detection method, the expression level of P27RF-Rho-siRNA+5-Fu cells in cancer associated protein P27 was significantly higher than that of the other 3 groups (P0.05), invasion related protein expression level of RhoC was lower than that of the other 3 groups (P0.05). Conclusion: P27RF-Rho gene silencing can significantly enhance the drug sensitivity of 5-Fu on SMMC7721 cells.

【作者單位】： 吉林大學(xué)第一醫(yī)院肝膽胰外科;
【基金】：吉林省衛(wèi)生廳自然科學(xué)基金資助課題(3D515U383428)
【分類號】：R735.7

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