RNA干扰Rac1基因表达抑制胃癌细胞增殖及促进凋亡的机制研究

Study on the mechanism of RNA interference Rac1 to inhibit the proliferation and promote apoptosis of human gastric cancer cell line

目的 RNA干扰抑制Rac1基因的表达对胃癌细胞增殖和凋亡的影响及机制。方法 Western blot检测胃癌组织及相应的癌旁组织中Rac1基因的蛋白表达;将NC-siRNA、Rac1-siRNA转染至人胃癌SGC-7901细胞,不经任何处理的细胞作为空白对照组,48 h后,Western blot检测各组细胞中Rac1的蛋白表达;CCK8实验检测细胞增殖;流式细胞仪检测细胞凋亡;Western blot检测ki67、PCNA、Cleaved caspase3、Notch1、Hes1蛋白表达。结果 Rac1在胃癌组织中的蛋白表达显著高于癌旁组织(P<0.01);RNA干扰SGC-7901细胞中Rac1基因后能显著降低Rac1的蛋白表达;与对照组及NC-siRNA组比较,Rac1-siRNA组细胞存活率及ki67、PCNA、Notch1、Hes1蛋白表达显著降低,细胞凋亡率及Cleaved caspase3蛋白表达显著升高(P<0.01)。结论 Rac1基因在胃癌组织中高表达,RNA干扰抑制Rac1基因表达后能显著降低细胞的增殖及诱导细胞凋亡,其机制与抑制Notch1信号通路有关。

Objective To investigate the effect of RNA interference Rac1 gene expression on the proliferation and apoptosis of gastric cancer cells.Methods The expression of Rac1 gene protein in gastric cancer tissues and adjacent tissues was detected by Western blot;NC-siRNA and Rac1-siRNA was transfected into human gastric cancer SGC-7901 cells,and without any treatment cells were used as blank control group,expression of Rac1 protein after 48h was detected by Western blot;cell proliferation was detected by CCK8 assay;cell apoptosis was detected by flow cytometry;ki67,PCNA,Cleaved caspase3,Notch1,Hes1 protein expression was detected by Western blot.Results The expression of Rac1 protein in gastric cancer tissues was significantly higher than that in paracancerous tissues （P＆lt;0.01）.RNA interference of Rac1 gene in SGC-7901 cells could significantly reduce the expression of Rac1 protein;compared with the control group and NC-siRNA group,cell survival rate and Ki67,PCNA,Notch1,Hes1 protein expression in Rac1-siRNA group was significantly decreased,cell apoptosis rate and the expression of Cleaved Caspase3 protein were significantly increased （P＆lt;0.01）.Conclusion Rac1 gene is highly expressed in gastric cancer tissues.RNA interference can inhibit the expression of Rac1 gene and decrease cell proliferation and induce apoptosis.The mechanism is related to the inhibition of Notch1 signaling pathway.