RNA干扰EP—CAM基因表达对胆囊癌细胞增殖及凋亡的影响

目的：利用RNA干扰技术抑制人胆囊癌GBC-SD细胞中上皮细胞黏附分子（EP-CAM）基因的表达,探讨抑制EP-CAM基因后对GBC-SD细胞增殖、凋亡的影响.方法：构建靶向EP-CAM基因的miRNA重组质粒,转染GBC-SD细胞,用RT-PCR和Western Blot分别检测EP-CAM基因的mRNA及蛋白的表达情况,MTT法检测细胞增殖活性,流式细胞仪检测细胞凋亡的变化.结果：靶向EP-CAM的miRNA重组质粒pcDNA^TM6.2-GW/EmGFPmiR-EPCAM-1构建成功,重组质粒能显著抑制靶基因EP-CAM的mRNA及蛋白表达.与对照组比较,干扰后的细胞凋亡率无明显变化（P＞0.05）,但细胞增殖活性受到明显抑制（P＜0.05）.结论：靶向EP-CAM基因的miRNA重组质粒能有效抑制EP-CAM基因的表达,抑制GBC-SD细胞的增殖.

Effects of EP-CAM Gene Silencing by RNA Interference on Proliferation and Apoptosis of Gallbladder Cancer Cells

Objective：To observe the effect of epithelial cell adhesion molecule （EP-CAM）gene silencing by RNA interfering technology on the proliferation and apoptosis of GBC-SD cells. Methods：Recombinant plasmid peDNA^TM6.2-GW/EmGFPmiR- EPCAM-1 targeting human EP-CAM gene was constructed and transfeeted into GBC-SD cells by lipofectamineTM 2000 after DNA sequencing. RT-PCR and western blot were used to detect the mRNA and protein expression of EP-CAM. After being tranfected by pcDNA^TM6.2-GW/EmGF- PmiR-EPCAM-1, cell proliferation was analyzed by MTT assay, apoptosis was detected by flow cytometry. Results：Recombinant plasmid pcDNA^TM6.2-GW/EmGFPmiR-EPCAM-1 targeting human EP-CAM gene was successfully constructed and efficiently suppressed the expression of EP-CAM mRNA and protein. The proliferation ability of the cells was reduced significantly. Compared with the control groups, the difference was significant（ P 〈 0.05 ）. However, cell apoptosis was not affected obviously（ P 〉 0.05 ）. Conclusions ：The recombinant plasmid pcDNA^TM6.2-GW/EmGFPmiR miRNA targeting human EP-CAM gene can effectively suppress EP-CAM expression, inhibit cellular proliferation in GBC-SD cells in vitro.