Effects of antisense-CXCR4 on the VEGF-C mRNA expression in colon cancer cell / 解放军医学杂志

ABSTRACT

Objective To investigate the effects of transfection of chemokine receptor CXCR4 antisense-RNA on the functional expression of VEGF-C mRNA and the invasive ability of colon cell line HT-29 in vitro.Methods PCR primers were designed according to the coding sequence of CXCR4 gene.EcoR Ⅰ and BamH Ⅰ recognition sequences and cutting sites were added to the 5' end of the antisense primer.The purified PCR fragment was retro-inserted into the cloning site of eukaryotic expression vector pcDNA3.1(+).The pcDNA3.1(+)for CXCR4 antisense-RNA was transfected into colon cancer HT-29 cells by liposome transfection.The expression of VEGF-C mRNA was detected by RT-PCR.The expression of CXCR4 protein was determined by Western blot.At the same time,cell growth kinetics was assessed by MTT assay,and in vitro invasive ability was assessed with Boyden chamber.Results CXCR4 antisense-RNA recombinant was successfully constructed.The expression of VEGF-C mRNA in antisense-CXCR4 transfected(HT-29tran)group decreased by 54.2%,while the expression of VEGF-C mRNA in vacant-transfected(HT-29KZ)group only decreased by 9.4% campared with non-transfected(HT-29)group.The difference between the two groups mentioned above was remarkable(P