摘要
目的利用人血管内皮生长因子-D(vascular endothelial growth factor-D,VEGF-D)小RNA干扰(siRNA),研究VEGF-D表达下调对人结肠癌细胞增殖和侵袭的影响。方法设立3个细胞组:未处理的人结肠癌SW480细胞作为空白对照组,转染阴性对照siRNA的SW480细胞作为阴性对照组,转染VEGF-D siRNA的SW480细胞作为实验组。RT-PCR和Western-blot检测转染后SW480细胞VEGF-D基因和蛋白的表达,MTT法检测VEGF-D RNA干扰对结肠癌细胞增殖的影响,Transwell小室实验检测VEGF-D RNA干扰对结肠癌细胞侵袭的影响。结果 VEGF-D siRNA转染人结肠癌细胞SW480,VEGF-D基因和蛋白水平表达量明显低于对照组,MTT实验显示体外细胞存活率下降,Transwell小室中穿膜细胞数减少。结论 VEGF-D siRNA显著抑制结肠癌细胞SW480中VEGFD的基因和蛋白表达,明显抑制SW480细胞的体外增殖和侵袭能力。
Objective To study the influence of downregulation of vascular endothelial growth factor-D (VEGF-D) on growth and invasion of human colonic carcinoma cells(SW480) by VEGF-D siRNA. Methods SW480 cells were divided into: untreated as blank group, transfected with negative controlled siRNA as negative control group, transfected with VEGF-D siRNA as experimental group. RT-PCR and Western-blot were used to study the expression of VEGF-D gene and protein, MTT was used to observe the influence of VEGF-D siRNA on cells growth, Transwell experiment was used to study the influence of VEGF-D siRNA on cells invasion. Results Expression levels of VEGF-D gene and protein were downregulated evidently after transfection of VEGF-D siRNA compared to controls, MTF showed that survival rate of the cells decreased, Transwell experiment showed that transmembrane eeUs number reduced in experiment group than in controls. Conclusion siRNA can downregulate the expression of VEGF-D evidently, and inhibit both growth and invasion ability of SW480 cells in vitro.
出处
《解剖科学进展》
CAS
2013年第5期416-419,共4页
Progress of Anatomical Sciences
基金
黑龙江省教育厅科技研究项目资助(No.12511217)