P27RF-Rho基因沉默对SMMC7721肝癌细胞5-Fu药物敏感性的实验研究

发布时间：2018-03-15 18:19

本文选题：肝癌　切入点：基因沉默　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨SMMC7721肝癌细胞P27RF-Rho基因沉默对于5-氟脲嘧啶(5-Fu)药物敏感性的影响,为临床晚期肝癌治疗提供理论依据。方法:构建P27RF-Rho RNAi载体,P27RF-Rho基因沉默慢病毒感染SMMC7721肝癌细胞,Western blotting法检测P27RF-Rho基因沉默效果,Western blotting法检测基因沉默效果。SMMC7721细胞分为:Scramble-si RNA阴性对照组、5-Fu组、P27RF-Rho-si RNA组和P27RF-Rho-si RNA+5-Fu组。MTT法检测各组细胞生长曲线;划痕实验检测细胞迁移能力;Transwell小室检测细胞侵袭能力;Western blotting法检测各组细胞中肿瘤相关蛋白P27及Rho C表达。结果:成功构建P27RF-Rho RNAi慢病毒载体。荧光显微镜检测慢病毒感染效果良好。Western blotting法显示P27RF-Rho-si RNA组P27RF-Rho蛋白表达明显低于SMMC7721组和Scramble-si RNA组,基因沉默效果良好。与3个对照组比较,P27RF-Rho-si RNA+5-Fu组细胞生长速度降低(P0.05);划痕实验中P27RF-Rho-si RNA+5-Fu组细胞迁移能力明显减低(P0.01);P27RF-Rho-si RNA+5-Fu组穿过Transwell小室微孔滤膜的平均细胞数明显少于另外三组(P0.01);Western blotting法检测P27RF-Rho-si RNA+5-Fu组癌相关蛋白P27表达水平明显高于其他三组(P0.05),侵袭相关蛋白Rho C表达水平低于其他三组(P0.05)。结论:P27RF-Rho基因沉默能明显增强5-Fu对肝癌SMMC7721细胞的药物敏感性。
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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