RNA干扰VEGF-C基因下调COX-2、Bcl-2表达对乳腺癌细胞凋亡的影响  

Vascular endothelial cell growth factor-C small RNA interfering vector down-regulate COX-2,Bcl-2 and induce apoptosis in human breast cancer cells

作　　者：顾斐[1] 邹强[1] 倪泉兴[1] 

机构地区：[1]复旦大学附属华山医院普外科,上海200040

出　　处：《复旦学报（医学版）》2009年第2期168-171,共4页Fudan University Journal of Medical Sciences

摘　　要：目的利用人血管内皮生长因子-C(vascular endothelial growth factor-C,VEGF-C)小RNA干扰(siRNA)表达载体——重组质粒pSilencer3.0-VEGF-C/siRNA,研究VEGF-C基因下调抑制COX-2、Bcl-2表达及对人乳腺癌细胞株MDA-MB-435的体外作用。方法pSilencer3.0-VEGF-C/siRNA表达质粒和阴性对照质粒稳定转染人乳腺癌细胞株MDA-MB-435,RT-PCR和Western blot检测转染前后VEGF-C、COX-2基因和VEGF-C、COX-2、Bcl-2蛋白的表达;MTT法和流式细胞仪检测VEGF-CRNA干扰对细胞的作用。结果pSilencer3.0-VEGF-C/siRNA转染乳腺癌细胞MDA-MB-435,VEGF-C基因和蛋白水平表达量明显降低,COX-2和Bcl-2表达下调,细胞体外活性下降,72h后凋亡率达60%。结论针对人VEGF-C基因的siRNA表达载体抑制VEGF-C表达的同时下调COX-2和Bcl-2的表达,促进乳腺癌细胞MDA-MB-435的凋亡。Objective To clone the recombinant plasmid silencing vascular endothelial growth factor- C （VEGF-C） gene by small interfering RNA （siRNA） and observe its role in the expression of COX-2, Bcl-2 and proliferation in human breast cancer cells MDA-MB-435. Methods Breast cancer cells MDA-MBA-435 were transfected stably with the recombinant siRNA plasmid and negative control by positive liposomal methods. We examined the gene expression of VEGF-C, COX-2 by RT-PCR and protein expression of VEGF-C, COX-2 and Bcl-2 by Western blot to explore the effect of VEGF-C down-regulation. The influences on cell activity and apoptosis were studied by MTT and flow cytometry. Results The down-regulation of VEGF-C inhibited the expression of some other genes such as COX-2 and bcl-2. The apoptosis rate was 60% after transfection 72 hours, and grew much slower than control groups. Conclusions The results obtained explain the function of VEGF-C and the cross-talk of downstream tyrosine kinase. Silencing of VEGF-C expression using siRNA vectors inhibits cell activity and induces apoptosis in human breast cancer cells MDA-MBA-435, which is a potential approach for tumor gene therapy.

关 键 词：乳腺癌 血管内皮生长因子-C 小RNA干扰 COX-2 BCL-2 

分 类 号：R737.9[医药卫生—肿瘤]