CD147siRNA对恶性黑色素瘤细胞VEGF及血管内皮细胞体外迁移活性的影响

Effects of CD147siRNA on VEGF Levels in Malignant Melanoma Cells and Migration Activity of Vascular Epithelial Cells in vitro

目的研究CD147siRNA对恶性黑色素瘤细胞株A375中VEGF表达水平和血管内皮细胞体外迁移活性的影响。方法采用半定量RT-PCR法检测转染后A375细胞中VEGF mRNA表达的变化。采用ELISA法检测A375细胞与成纤维细胞共同培养后VEGF表达水平的变化。采用细胞迁移实验检测转染CD147siRNA前后的A375细胞对脐静脉内皮细胞株ECV-304在体外迁移活性的影响。结果转染CD147siRNA后,A375细胞中VEGF的mRNA和蛋白表达水平显著降低,其下调率分别为10.77%(P<0.05)和38.5%(P<0.01)。转染CD147siRNA前后的A375细胞与成纤维细胞共同培养后VEGF的表达水平均较单独培养时显著升高,其升高率分别为98.5%(P<0.01)和159.9%(P<0.01)。转染CD147siRNA后的A375细胞使ECV-304细胞的体外迁移活性下降44.77%(P<0.01)。结论 CD147siRNA能有效抑制恶性黑色素瘤细胞株A375中VEGF的表达水平和脐静脉内皮细胞株ECV-304在体外的迁移活性。

Objective To investigate the effects of CD147siRNA on the VEGF levels in malignant melanoma cell line A375 and the migration activity of vascular epithelial cells in vitro.Methods The change of the expression of VEGF mRNA in transfected A375 cells was identified by semi-quantitive RT-PCR.The change of the expression of soluble VEGF in coculture medium of A375 cells with fibroblasts was detected by ELISA assay.The effect of A375 cells transfected with CD147siRNA on migration activity of human umbilical vein epithelial cell line ECV-304 in vitro was evaluated by endothelial cell migration assay.Results The VEGF mRNA and protein levels significantly down-regulated after A375 cells were transfected with CD147siRNA,the reduction rates were 10.77%（P0.05） and 38.5%（P0.01） respectively.The expression of soluble VEGF in coculture medium of A375 cells with fibroblasts and of C1 cells with fibroblasts both significantly up-regulated as compared with tumor cells single cultured,the increase rates were 98.5%（P0.01） and 159.9%（P0.01） respectively.The transfected A375 cells reduced the migration activity of ECV-304 cells for 44.77%（P0.01）.Conclusion CD147siRNA could effectively inhibit the VEGF levels in malignant melanoma cell line A375 and the migration activity of vascular epithelial cells in vitro.