特异siRNA沉默Slug基因抑制体外胰腺癌AsPC-l细胞生长的实验研究

Effects of Slug gene silence by siRNA on proliferation and apoptosis of AsPC-1 in vitro

目的:研究Slug基因沉默对胰腺癌AsPC-l细胞增殖、凋亡的影响。方法:构建靶向抑制S1ug的siRNA表达载体,瞬时转染AsPC-1细胞。采用RT-PCR和Westernblot法检测细胞Slug蛋白,MTT检测细胞增殖,AnnexinV-FITC/PI了解细胞凋亡的变化。结果:成功构建了pGenesil-l-Slug-siRNA(pSlug-siRNA)和pCenesil-1-Neg-siRNA(pNeg-siRNA)表达质粒。pSlug-siRNA瞬时转染AsPC-1细胞72h后,细胞S1ug表达及生长受到明显抑制,细胞凋亡率明显增加。结论:Slug-siRNA抑制Slug表达,促进As-PC-1细胞凋亡并抑制细胞的增殖。

Objective： To inhibit Slug expression in pancreatic cancer cells with small interfering RNA （siRNA）, and explore its effect on cell proliferation and apoptosis. Methods：Plasmaid S pSlug-siRNA and pNeg- SiRNA（negative control） were constructed and transfected into AsPC-1 cells. The expression of Slug was detected by RT-PCR and Western blotting. The proliferation of CaPan-2 cells in vitro was determined by MTT assay. The apoptotic rates of the cells were detected by flow cytometry. Results：Two cell clones were screened from pSlug-siRNA and pNeg-siRNA-transfected AsPC-1 cells. Slug expression in AsPC-1 cells was suppressed markedly after transfection of pSlug-siRNA. The apoptosis rate was significantly higher in pSlug-siRNA cells than in pNeg-siRNA cells. Apoptosis could be seen more easily by Hoechst 33258 and PI staining at 72h in pSlug-siRNA cells than in pNeg-siRNA and control cells. Conclusion ： Slug inhibition can inhibit the proliferation of AsPC-1 cell in vitro, and accelerate the apoptosis.