本文選題：肝癌　切入點(diǎn)：基因沉默　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討SMMC7721肝癌細(xì)胞P27RF-Rho基因沉默對于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P27RF-Rho基因沉默效果,Western blotting法檢測基因沉默效果。SMMC7721細(xì)胞分為:Scramble-si RNA陰性對照組、5-Fu組、P27RF-Rho-si RNA組和P27RF-Rho-si RNA+5-Fu組。MTT法檢測各組細(xì)胞生長曲線;劃痕實(shí)驗(yàn)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Rho C表達(dá)。結(jié)果:成功構(gòu)建P27RF-Rho RNAi慢病毒載體。熒光顯微鏡檢測慢病毒感染效果良好。Western blotting法顯示P27RF-Rho-si RNA組P27RF-Rho蛋白表達(dá)明顯低于SMMC7721組和Scramble-si RNA組,基因沉默效果良好。與3個對照組比較,P27RF-Rho-si RNA+5-Fu組細(xì)胞生長速度降低(P0.05);劃痕實(shí)驗(yàn)中P27RF-Rho-si RNA+5-Fu組細(xì)胞遷移能力明顯減低(P0.01);P27RF-Rho-si RNA+5-Fu組穿過Transwell小室微孔濾膜的平均細(xì)胞數(shù)明顯少于另外三組(P0.01);Western blotting法檢測P27RF-Rho-si RNA+5-Fu組癌相關(guān)蛋白P27表達(dá)水平明顯高于其他三組(P0.05),侵襲相關(guān)蛋白Rho C表達(dá)水平低于其他三組(P0.05)。結(jié)論:P27RF-Rho基因沉默能明顯增強(qiáng)5-Fu對肝癌SMMC7721細(xì)胞的藥物敏感性。
[Abstract]:Objective: to investigate the effect of P27RF-Rho gene silencing on the drug sensitivity of 5-fluorouracil (5-Fu) in SMMC7721 hepatoma cells. Methods: P27RF-Rho RNAi vector was constructed to silence lentivirus infected SMMC7721 hepatoma cells. Western blotting assay was used to detect the effect of P27RF-Rho gene silencing. SMMC7721 cells were divided into: Scramble-si RNA. The growth curves of P27RF-Rho-si RNA group and P27RF-Rho-si RNA 5-Fu group were detected by MTT assay. Western blotting assay was used to detect the expression of tumor associated protein P27 and Rho C. Results: P27RF-Rho RNAi lentivirus vector was successfully constructed. Fluorescence microscopy was used to detect chronic disease. The expression of P27RF-Rho protein in P27RF-Rho-si RNA group was significantly lower than that in SMMC7721 group and Scramble-si RNA group. The cell growth rate of P27RF-Rho-si RNA 5-Fu group was significantly lower than that of three control groups, and the cell migration ability of P27RF-Rho-si RNA 5-Fu group was significantly lower than that of P27RF-Rho-si RNA 5-Fu group, and the average cell number of P27RF-Rho-si RNA 5-Fu group through Transwell micropore filter membrane was significantly decreased in scratch test. The expression of P27RF-Rho-si RNA 5-Fu in the P27RF-Rho-si RNA 5-Fu group was significantly higher than that in the other three groups, and the expression of invasive-associated protein Rho C was lower than that in the other three groups. Conclusion the silencing of P27RF-Rho gene can significantly enhance the expression of 5-Fu on SMMC7721 of hepatocellular carcinoma. The drug sensitivity of cells.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.7

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