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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 investigate the effect of VEGF-D expression on the proliferation and invasion of human colon cancer cells by using small interfering RNA (siRNA) of human vascular endothelial growth factor-D (VEGF-D). Methods Three cell lines were established: untreated human colon cancer SW480 cells as control group, SW480 cells transfected negative control siRNA as negative control group and SW480 cells transfected with VEGF-D siRNA as experimental group. The expression of VEGF-D gene and protein in SW480 cells was detected by RT-PCR and Western-blot. The effect of VEGF-D RNA interference on the proliferation of colon cancer cells was detected by MTT assay. The effect of VEGF-D RNA interference on the proliferation of colon cancer Impact of cell invasion. Results The expression of VEGF-D siRNA and VEGF-D mRNA and protein in human colon cancer cell lines SW480 and VEGF-D were significantly lower than those in control group. The MTT assay showed that the cell viability decreased in vitro and the number of transmembrane cells in Transwell cells decreased. Conclusion VEGF-D siRNA significantly inhibits VEGFD gene and protein expression in SW480 cells and significantly inhibits the proliferation and invasion of SW480 cells in vitro.