Nucleostemin基因特异性RNA干扰对胃癌细胞增殖和凋亡的影响

The effect on proliferation and apoptosis by Nucleostemin gene specific RNA interference in gastric-carcinoma cells

目的探讨应用RNA干扰技术抑制nucleostemin(NS)基因表达对胃癌SGC-7901细胞株增殖和凋亡的影响。方法阳离子脂质体法将构建好的NS-siRNA表达载体转染胃癌SGC-7901细胞株,采用MTT法测试各组细胞增殖抑制率;RT-PCR法检测各组细胞NS基因表达情况,流式细胞术检测细胞周期,Annexin-V-FITC/PI法检测细胞凋亡率。结果与对照组相比,S组细胞的增殖减慢,24、48、72 h细胞增殖抑制率分别为53.21%、71.54%、87.47%;NS基因表达量下降,G0/G1期细胞百分率显著升高(58.34%),S期细胞减少(20.68%),细胞凋亡率增加(26.85%)。结论RNA干扰技术可抑制胃癌SGC-7901细胞株NS基因的表达,使细胞增殖减慢,细胞周期阻滞在G0/G1期,凋亡率增加。

Objective To investigate the effect on proliferation and apoptosis by RNA interference to inhibit Nucleostemin （NS） gene expression in gastric carcinoma SGC-7901 ceils. Methods The NS-siRNA expression vector was transfected into gastric-carcinoma cells with LipofectamineTM 2000 reagent. Then we detected the cell proliferation inhibition ratio by MTT assay, the levels of NS gene expression in all groups by RT-PCR, cell cycle by flow cytometry, cell apoptosis ratio by Annexin-V-FITC/PI kit. Results Compared with that in the control group, cell proliferation in S group decreased; at 24, 48 and 72 h the cell proliferation inhibition ratio was 53.21%, 71.54% and 87. 47%, respectively, the level of NS gene expression reduced in S group. G0/G1 phase cell was 58.34%, S phase cell was 20.68%, and the cell apoptosis was 26.85%. Conclusion RNA interference could substantially inhibit NS gene expression in gastric carcinoma SGC-7901 cells, decrease cell proliferation, arrest cell cycle and increase apoptosis ratio.