siRNA靶向沉默TET2基因对人肝癌细胞增殖和凋亡的影响  

Effects of targeted silencing of TET2 gene by si RNA on cell proliferation and apoptosis of human hepatic cancer cells

作　　者：景晓乾 杨卫平[1] 刘心玉[1] 汪炳瑞 马丁[1] 邱伟华[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院外科上海消化外科研究所,上海200025

出　　处：《外科理论与实践》2015年第4期338-342,共5页Journal of Surgery Concepts & Practice

基　　金：国家自然科学基金(81172326)

摘　　要：目的:通过siRNA靶向沉默TET2(ten-eleven translocation 2)基因,以研究TET2基因对于人肝癌细胞增殖和凋亡的影响并初步探讨其可能机制。方法:RT-PCR和Western印迹法检测人肝癌细胞株和正常人肝星状细胞LO2中TET2表达情况。siRNA转染入Bel-7404和SMMC-7721肝癌细胞内,并设置阴性对照组(NC组)。靶向沉默TET2基因后,CCK-8法检测肝癌细胞增殖能力,FITC AnnexinⅤ-Apoptosis Detection KitⅠ检测肝癌细胞的凋亡;Western印迹法检测下游信号通路中Caspase-3和Caspase-8蛋白表达。结果:与正常人肝星状细胞LO2相比,TET2基因在人肝癌细胞中高表达。siRNA转染入SMMC-7721和Bel-7404细胞后,TET2基因的表达显著降低。与NC组相比,转染48 h后肝癌细胞的增殖水平显著降低(P<0.05)。肝癌细胞的凋亡水平率显著升高,下游信号通路中,Capase-3蛋白和Capase-8蛋白的表达量显著上调。结论:siRNA靶向沉默TET2基因可通过促进肝癌细胞凋亡的发生抑制其增殖。Objective To investigate the effects of TET2（ten-eleven translocation 2） gene on the proliferation and apoptosis of human hepatic cancer cells and explore the possible mechanisms using targeted silencing of TET2 by siRNA.Methods RT-PCR and Western blot were used to detect the expression of TET2 gene on human hepatic cancer cell line and human hepatic stellate cell LO2. siRNA was transfected into hepatic cancer cells Bel-7404 and SMMC-7721, as study group and negative control（NC） group was set. After the targeted silencing of TET2 gene, CCK-8 method was used to detect the proliferation of hepatic cancer cells, and FITC Annexin Ⅴ-Apoptosis Detection Kit Ⅰ to detect the apoptosis of them.Western blot was used to detect the expression of downstream signal pathway proteins including Caspase-3 and Caspase-8.Results The expression of TET2 gene in human hepatic cancer cells was higher significantly than that in human hepatic stellate cells LO2. The expression of TET2 gene in the cells SMMC-7721 and Bel-7404, which si RNA was transfected into,was decreased significantly. Compared with NC, the proliferation was suppressed after 48 h transfection（ P〈0.05）. The incidence of cell apoptosis increased significantly. The expressions of Caspase-3 and Caspase-8 were increased significantly.Conclusions si RNA targeted silencing TET2 gene could inhibit the proliferation of hepatic cancer cells by promoting the apoptosis.

关 键 词：肝细胞癌 TET2基因 小干扰RNA 增殖 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]