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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 effect of CD147 siRNA on the expression of VEGF and the migration of vascular endothelial cells in vitro in the malignant melanoma cell line A375. Methods The expression of VEGF mRNA in A375 cells was detected by semi-quantitative RT-PCR. The expression of VEGF in A375 cells and fibroblasts were detected by ELISA. The effect of A375 cells transfected with CD147 siRNA on the migration of human umbilical vein endothelial cell line ECV-304 in vitro was tested by cell migration assay. Results After transfection with CD147 siRNA, the mRNA and protein expressions of VEGF in A375 cells were significantly decreased, with the down-regulation rates of 10.77% (P <0.05) and 38.5% (P <0.01), respectively. The expression of VEGF in A375 cells transfected with CD147 siRNA before and after transfection was significantly higher than that cultured alone (98.5%, P <0.01) and 159.9% (P <0.01), respectively. A375 cells transfected with CD147 siRNA reduced ECV-304 cells migration in vitro by 44.77% (P <0.01). Conclusion CD147 siRNA can effectively inhibit the expression of VEGF in human melanoma cell line A375 and the migration of ECV-304 in vitro.