Livin靶向RNA干扰对人胃癌细胞SGC7901细胞增殖和凋亡的影响

Livin-targeted siRNA inhibits the growth of SGC7901 cells and induces apoptosis

目的探讨Livin靶向RNA干扰对SGC7901细胞株生物学特性的影响,探索胃癌基因治疗的新途径。方法设计和构建Livin特异性的siRNA真核表达载体,转染SGC7901细胞,应用RT-PCR分析LivinmRNA的表达情况,采用MTT法检测细胞的生长状况,通过流式细胞仪进行细胞凋亡的检测。结果构建真核表达载体p-siRNA1并转染SGC7901细胞后,RT-PCR结果显示2条Livin siRNA真核表达质粒均能有效抑制Livin mRNA的转录表达,干扰组细胞与未转染组细胞比较,生长受到明显抑制,凋亡率明显增加。结论成功构建了Livin干扰真核表达载体,siRNA重组体能有效抑制人胃癌细胞Livin mRNA表达,并抑制胃癌细胞生长,促进其凋亡。

Objective To study he effects of Livin - targeting siRNA on the growth and apoptosis of SGC7901 cells and to explore a new pathway for gastric carcinoma genetherapy. Methods Two pairs of DNA template coding siRNA against Livin were synthesized and cloned into the vector pGenSil - 1 which was transfected into SGC7901 cells. The expression of survivin mRNA was detected by semi -quantitive RT- PCR. MrfT assay was applied to determine the cell growth status. Apoptotic rates were evaluated by FCM. Results DNA sequencing results all showed that the 2 target segments were cloned into pGenSil - 1 vector respectively. After transfected into SGC7901 cells, RT - PCR showed the expression of Livin mRNA was down - regulated in the transfected cells（ P 〈 0.05 ）. The proliferation of SGC7901 cells was obviously inhibited and the apoptotic rates were significantly increased （ P 〈 0.05 ）. Conclusion The siRNA eukaryotic expression vector of Livin have been constructed successfully which significantly inhibit Livin expression, suppress the growth of SGC7901 cells and induce apotosis.