摘要
目的:探讨miR-126对EA.hy926细胞血管内皮生长因子(VEGF)表达的影响。方法:选用EA.hy926细胞株作为研究对象,采用阳离子介导的转染方式,将miR-126 inhibitor和miR-126 mimics进行细胞转染,分析miR-126对血管内皮细胞功能的影响。采用转染negative control作为阴性对照,转染36 h后提取细胞总RNA,利用real-time PCR和Western blotting分别检测EA.hy926细胞VEGF mRNA和蛋白水平的表达。结果:转染miR-126 inhibitor 50 nmol/L 36 h后,EA.hy926细胞的VEGF mRNA及蛋白水平各组间差异有统计学意义(P<0.01),与阴性对照组相比,miR-126 inhibitor使EA.hy926细胞的VEGF mRNA及蛋白水平显著上调(P<0.01);转染miR-126 mimics 50 nmol/L 36 h后,EA.hy926细胞的VEGF mRNA及蛋白水平各组间有显著差异(P<0.01),与阴性对照组相比,miR-126 mimics使EA.hy926细胞的VEGF在mRNA及蛋白水平显著下调(P<0.01)。结论:miR-126能够抑制VEGF的表达,VEGF有可能是其调节的靶基因之一。
AIM： To explore the effects of miR-126 on the expression of vascular endothelial growth factor （VEGF） in vascular endothelial cell line EA. hy926. METHODS： EA. hy926 cells were cultured in vitro and transfected with miR-126 mimics or miR-126 inhibitor by cation-mediated transfection method. The total RNA was extracted from the culture cells 36 h after transfection of miR-126 mimics or miR-126 inhibitor. The expression of VEGF at mRNA and protein levels was detected by real-time PCR and Western blotting. RESULTS ： Thirty-six hours after transfection of miR-126 in- hibitor at concentration of 50 nmol/L, the expression of VEGF at mRNA and protein levels increased significantly as com- pared with the negative control （ P 〈 0. 01 ）. However, transfection of miR-126 mimics at concentration of 50 nmol/L for 36 h significantly decreased the expression of VEGF at mRNA and protein levels as compared with the negative control. CON- CLUSION： miR-126 inhibits the expression of VEGF. VEGF may be one of the target genes regulated by miR-126.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第1期30-34,共5页
Chinese Journal of Pathophysiology
