miR-210对血管内皮细胞VEGF-Notch信号通路的调控  

Regulatory effect of miR-210 overexpression on VEGF-Notch signaling pathway

作　　者：娄远蕾[1] 刘芬[1] 高法梁[2] 阮琼芳[1] 崔苏萍[1] 邓志锋[2] 汪泱[1] 

机构地区：[1]南昌大学第一附属医院泌尿外科研究所,江西南昌330006 [2]南昌大学第二附属医院神经外科,江西南昌330006

出　　处：《中国病理生理杂志》2011年第5期923-927,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30960396;No.81060059)

摘　　要：目的:观察过表达miR-210对血管内皮细胞中血管内皮生长因子(VEGF)-Notch信号通路相关分子表达的影响,探讨miR-210调控血管新生的分子机制。方法:采用常规方法培养人脐静脉内皮细胞(HUVE-12)。通过转染LV-miR-210-GFP重组慢病毒载体上调内皮细胞miR-210表达,实验分为miR-210过表达组(LV-miR-210-GFP)和对照组(LV-GFP)。采用real-time PCR检测miR-210的表达变化,流式细胞术检测ephrin-A3的表达,采用real-time PCR、Western blotting、免疫荧光细胞化学染色法分别检测VEGF-Notch信号通路相关分子VEGF、VEGF 2(VEGFR 2)、Notch1的mRNA和蛋白表达水平。结果:Real-time PCR结果显示,与对照组相比,miR-210过表达组miR-210表达水平上调了(32.10±1.26)倍;流式细胞术结果显示,miR-210过表达组阳性细胞率[(73.22±1.45)%]较对照组[(12.52±0.67)%]明显增加(P<0.05)。Real-time PCR结果显示,miR-210过表达组VEGF、VEGFR2、Notch1 mRNA分别较对照组上调了(7.40±0.67)、(2.50±0.10)、(9.70±0.72)倍,P<0.05;免疫荧光结果显示,miR-210过表达组VEGF和VEGFR2红色荧光信号均显著强于对照组(P<0.05)。Western blotting结果显示,miR-210过表达组血管内皮细胞中Notch1的蛋白表达量(4.22±0.60)较对照组明显升高(P<0.05)。结论:miR-210过表达可显著上调VEGF-Notch信号通路分子的表达水平。AIM： To explore the potential mechanism of angiogenesis by investigating the regulatory effect of miR-210 overexpression on vascular endothelial growth factor（VEGF）-Notch signaling pathway.METHODS： MiR-210 overexpression was induced in human umbilical vein endothelial（HUVE-12） cells by lentivirus transfection.The HUVE-12 cells were divided into 2 groups： miR-210 overexpression group（LV-miR-210-GFP） and control group（LV-GFP）.The change of ephrin-A3 protein was detected by flow cytometry analysis.The mRNA and protein levels of VEGF-Notch signaling molecules,VEGF,VEGF receptor 2（VEGFR 2）and Notch 1,were determined by the methods of real-time PCR,immunofluorescence and Western blotting.RESULTS： Compared to the control cells,significant down-regulation of ephrin-A3 protein was observed in HUVE-12 cells with miR-210 overexpression,while both mRNA and protein levels of VEGF,VEGFR2 and Notch1 were up-regulated.CONCLUSION： miR-210 may be involved in VEGF-Notch signaling pathway to regulate angiogenesis.

关 键 词：MIR-210 血管内皮细胞 血管内皮生长因子 NOTCH1 

分 类 号：R363[医药卫生—病理学]