si-RNA介导FGFR1基因沉默对胃癌细胞生长及凋亡的影响

Effect of siRNA-mediated FGFR1 gene silencing on growth and apoptosis of gastric cancer cell line SGC-7901

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 si RNA-induced FGFR1 gene down-regulation on proliferation and apoptosis of gastric cancer SGC-7901 cells. Methods si RNA-FGFR1 and the negative-control(NC)-si RNA 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 si RNA-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 si RNA-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 si RNA-FGFR1 in comparison to that in NC-si RNA and mock groups. Flow cytometry and Hoechst staining showed that the apoptotic rate was significantly increased in si RNA-FGFR1 transfected cells compared to control cells. Conclusion si RNA-FGFR1 can significantly inhibit FGFR1 expression, suppress the growth and induce apoptosis in FGFR1-overexpressing gastric cancer SGC-7901 cells.