转染Chk1 siRNA的人胃癌SGC7901细胞对顺铂的敏感性

摘要

Aim: To investigate the influence of cisplatin on proliferation and apoptosis of human gastric cancer SGC7901 cells transfected with Chk1 siRNA.Methods:Three kinds of Chk1 siRNA were established and transfected into SGC7901 cells.Real-time PCR and Western blot were used to detect the expressions of Chk 1 mRNA and protein in SGC7901 cells, respectively.The Chk1 siRNA with strongest inhibitory effect on Chk 1 expression was transfected into SGC7901 cells for 24 h, then treated by cisplatin(10μmol/L) for 12 h, MTT method was used to detect the cell prolifera-tion, and TUNEL method was used to detect cell apoptosis .The cells treated with cisplatin alone and without any treatment were the controls.Results:Chk1 protein and mRNA expression levels in SGC7901 cells transfected with Chk1 mRNA both decreased(P<0.05), and the inhibition effect of Chk1 S2 was stronger.The cell proliferation inhibition rate and apoptosis index in Chk1 S2+cisplatin group were higher than those in the 2 control groups(P<0.05).Conclusion:Silencing Chk1 expression might enhance the sensitivity of SGC 7901 cells to cisplatin .%目的:探讨Chk1 siRNA对顺铂作用的人胃癌SGC7901细胞增殖及凋亡的影响.方法:构建3个Chk1 siRNA重组载体,分组转染人胃癌SGC7901细胞,采用实时荧光定量PCR方法检测细胞中Chk1 mRNA的表达,应用Western blot检测Chk1蛋白的表达.以效应最强的Chk1 siRNA转染胃癌SGC7901细胞24 h,再用终浓度为10μmol/L的顺铂处理12 h(Chk1 S2+顺铂组).运用MTT法检测细胞增殖抑制率,TUNEL法检测细胞凋亡.以顺铂处理和未经任何处理的细胞作对照.结果:3条Chk1 siRNA转染SGC7901细胞后,Chk1 mRNA及蛋白表达减弱(P<0.05),以Chk1 S2效应最佳.Chk1 S2+顺铂组SGC7901细胞的增殖抑制率和凋亡指数(AI)均高于顺铂组及对照组(P<0.05).结论:体外沉默Chk1表达可增强人胃癌SGC7901细胞对顺铂的敏感性.