siRNA干扰沉默GS基因抑制胃癌细胞生长的研究

Experimental study on silencing the GS expression by siRNA to inhibit the growth of gastric cancer cells

目的:利用RNA干扰技术,研究靶向Glutamine Synthetase(GS)基因的小干扰RNA(siRNA)在体外对胃癌细胞生长的影响,探索胃癌基因治疗并了解GS在胃癌发生、发展中的作用。方法:合成GSsiRNA,并用脂质体法将GSsiRNA转至胃癌BGC-823细胞中;采用实时定量PCR和Western印迹法观察GSsiRNA转染前后胃癌细胞GS基因及相应蛋白表达的变化;用CCK8、流式细胞检测技术分别检测胃癌细胞增殖及凋亡的变化。结果:转染GSsiRNA后的胃癌细胞BGC-823与对照组相比,生长明显变缓(P<0.05),细胞凋亡率明显增高(P<0.05)。结论:靶向GSsiRNA可明显下调靶基因GS的表达,在体外可抑制胃癌BGC-823细胞的生长并促进其凋亡。

Objective To determine the inhibitory effect of synthetic GS short interfering RNA（siRNA） on the expression of GS gene in human gastric cancer line BGC-823 cells;and to elucidate its effect on the proliferation of BGC823 cells in vitro,in order to probe the feasibility of gene therapy in this domain.Methods siRNA targeting GS mRNA was transfected into BGC-823 cells,and GS expression was determined by real-time PCR and Western blot.Cell proliferation and apoptosis were examined by CCK8 and flow cytometry,respectively.Results GS siRNA significantly inhibited the expression of GS in gastric cancer cells at both mRNA and protein levels;the rate of apoptosis of BGC-823 cells was significantly different from that of the control groups（P〈0.05）.The proliferation of BGC-823 cells was also significantly suppressed in vitro（P 〈0.05）.Conclusions siRNA directed against GS can significantly suppress the proliferation of BGC-823 cells in vitro.This might provide a new approach of gene therapy for gastric cancer.