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miR-126对EA.hy926细胞血管内皮生长因子表达的调控作用

miR-126 regulates expression of vascular endothelial growth factor in EA.hy926 cells

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【作者】 郑志伟朱志伟郑滢陈蕾

【Author】 ZHENG Zhi-wei;ZHU Zhi-wei;ZHENG Ying;CHEN Lei;Department of Pharmacy,Cancer Hospital of Shantou University Medical College;Department of Medicine,Cancer Hospital of Shantou University Medical College;

【机构】 汕头大学医学院附属肿瘤医院药学部汕头大学医学院附属肿瘤医院内科

【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。

【Abstract】 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 inhibitor at concentration of 50 nmol / L,the expression of VEGF at mRNA and protein levels increased significantly as compared 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.CONCLUSION:miR-126 inhibits the expression of VEGF.VEGF may be one of the target genes regulated by miR-126.

  • 【文献出处】 中国病理生理杂志 ,Chinese Journal of Pathophysiology , 编辑部邮箱 ,2014年01期
  • 【分类号】R363
  • 【被引频次】2
  • 【下载频次】151
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