转染Chk1 siRNA的人胃癌SGC7901细胞对顺铂的敏感性

Sensitivity of human gastric cancer SGC7901 cells transfected with Chk1 siRNA 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。

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 Chk1 mRNA and protein in SGC7901 cells,respectively. The Chk1 siRNA with strongest inhibitory effect on Chk1 expression was transfected into SGC7901 cells for 24 h,then treated by cisplatin（ 10 μmol/L） for 12 h,MTT method was used to detect the cell proliferation,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 SGC7901 cells to cisplatin.