应用shRNA技术研究NDRG2在人胃癌细胞SGC7901增殖中的作用

Study of the role of NDRG2 in the proliferation of SGC7901 cell by shRNA-mediated RNA interference

目的:构建针对人 N-myc 下游调节基因2(NDRG2)的短发夹 RNA(shRNA)表达载体 pSNAV-shRNA,观察其在SGC7901细胞中对 NDRG2的抑制作用以及 NDRG2被抑制后细胞增殖能力的变化。方法:PCR 扩增针对 NDRG2的 shRNA 表达框架,构建 pSNAV-shRNA 质粒,转染 SGC7901细胞,通过 RT-PCR,免疫印迹试验检测其对 NDRG2表达的影响;通过细胞周期分析,软琼脂集落形成试验检测细胞增殖能力的变化。结果:将构建好的 pSNAV-shRNA 质粒转染 SGC7901细胞后,NDRG2的表达受到明显的抑制,细胞周期 G1期阻滞,集落形成能力降低。结论:构建的 pSNAV-shRNA 质粒能够有效抑制NDRG2在 SGC7901细胞中的表达,降低 SGC7901细胞的增殖能力。

Objective：To construct an expression vector carrying a short hairpin RNA （shRNA）against human N - myc downstream regulated gene 2 （NDRG2） to suppress the expression of NDRG2 in SGC7901 cell and test the change of cell proliferation. Method：Plasmid pSNAV - shRNA carrying shRNA aiming at NDRG2 was transfected into SGC7901 cell and its effect on NDRG2 expression was studied by RT - PCR and west blotting. The change of cell proliferation was studied by cell cycle analysis and colony formation assay. Results：The recombinant plasmid pSNAV - shRNA was successfully constructed. The recombinant plasmids suppressed the expression of NDRG2 sig- nificantly and cell proliferation reduced accordingly. Conclusion：The shRNA of NDRG2 can inhibit the expression of NDRG2 efficiently in SGC7901 cell and reduce SGC7901 cell＇s ability of proliferation.