si-RNA介导FGFR1基因沉默对胃癌细胞生长及凋亡的影响

摘要

目的 探索靶向沉默成纤维细胞生长因子受体1(FGFR1)表达对胃癌细胞SGC-7901生长及凋亡的影响.方法 通过Western blotting法研究FGFR1蛋白在胃癌细胞SGC-7901以及人正常胃黏膜上皮细胞GES-1的表达情况;设计合成特异性针对FGFR1基因的小干扰RNA(siRNA)及阴性对照-siRNA(NC-siRNA);采用瞬时转染法将siRNA转入胃癌细胞SGC-7901中,Western blotting法检测转染siRNA-FGFR1后对SGC-7901细胞中FGFR1蛋白表达的影响.通过MTT法检测siRNA-FGFR1对SGC-7901细胞增殖抑制的时间-效应关系,以及克隆集落形成实验检测siRNA-FGFR1对SGC-7901细胞克隆集落形成的影响;采用Hoechst染色法和流式细胞术检测siRNA-FGFR1对胃癌细胞凋亡的影响.结果 与人正常胃黏膜上皮细胞GES-1相比,胃癌细胞SGC-7901的FGFR1蛋白表达水平明显增高(P＜0.05);siRNA-FGFR1转染SGC-7901细胞后,FGFR1蛋白表达水平明显下调(P＜0.05);胃癌细胞增殖生长曲线提示,用siRNA技术沉默胃癌细胞中FGFR1的表达后,相较于空白组和NC-siRNA组,siRNA-FGFR1组细胞增殖明显受抑制,另外胃癌细胞SGC-7901克隆集落形成明显受抑制;流式细胞术结果表明,正常组与NC-siRNA组中胃癌SGC-7901细胞早期凋亡百分比差异无统计学意义(P＞0.05),而siRNA-FGFR1组细胞早期凋亡百分比显著升高(P＜0.01).结论 siRNA-FGFR1可抑制FGFR1蛋白在人胃癌细胞SGC-7901中的表达,并抑制胃癌细胞的生长,促进其凋亡.以FGFR1为靶点的小RNA干扰技术有望成为胃癌靶向治疗的新方法.%Objective To investigate the effect of siRNA-induced FGFR1 gene down-regulation on proliferation and apoptosis of gastric cancer SGC-7901 cells.Methods siRNA-FGFR1 and the negative-control(NC)-siRNA were designed and synthesized,then transfected into SGC-7901 cells.The expression levels of FGFR1 protein was detected by Western blotting analysis.The MTT assay and cell clonogenic assay were used to investigate the inhibitory effect of siRNA-FGFR1 on the viability of gastric cancer SGC-7901 cells.The apoptosis of SGC-7901 cells was detected by flow cytometry with Hoechst staining.Results The expression of FGFR1 in SGC-7901 cells was higher than that in human normal gastric mucosa epithelial GES-1 cells (P＜0.05),and the expression level of FGFR1 in SGC-7901 cells after transfection with siRNA-FGFR1 was down-regulated (P＜0.01).The MTT and clonogenic assay showed that the growth of SGC-7901 cells was inhibited after transfecton with siRNA-FGFR1 in comparison to that in NC-siRNA and mock groups.Flow cytometry and Hoechst staining showed that the apoptotic rate was significantly increased in siRNA-FGFR1 transfected cells compared to control cells.Conclusion siRNA-FGFR1 can significantly inhibit FGFR1expression,suppress the growth and induce apoptosis in FGFR1-overexpressing gastric cancer SGC-7901 cells.