文档介绍：Nucleostemin基因特异性RNA干扰对胃癌细胞增殖和凋亡的影响
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作者:郑学芝,胡静,孙卫,刘桂莲,刘晓霓,蔡子微
【摘要】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、%、%、%;NS基因表达量下降,G0/G1期细胞百分率显著升高(%),S期细胞减少(%),细胞凋亡率增加(%)。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。
【关键词】 RNA干扰;nucleostemin;胃癌;细胞增殖;凋亡
ABSTRACT: Objective To investigate the effect on proliferation and apoptosis by RNA interference to inhibit Nucleostemin (NS) gene expression in gastric carcinoma SGC
7901 cells. Methods The NSsiRNA expression vector was transfected into gastriccarcinoma cells with M2000 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. pared with that in the control group, cell proliferation in S group decreased; at 24, 48 and 72h the cell proliferation inhibition ratio was %, % and %, respectively, the level of NS gene expression reduced in S group. G0/G1 phase cell was %, S phase cell was %, and the cell apoptosis was %. Conclusion RNA interference could substantially inhibit NS gene exp