miR-137靶向Notch1调控人视网膜微血管内皮细胞的增殖、周期和凋亡  

Effects of miR-137 Regulating Notch1 on Cycle and Apoptosis of Human Retinal Microvascular Endothelial Cells

作　　者：张亨闰 周春兰[1] 陈红梅[1] 张静[1] 康海军 Zhang Hengrun;Zhou Chunlan;Chen Hongmei(Department of Ophthalmology Suining Central Hospital,Suining,Sichuan 629000,China)

机构地区：[1]遂宁市中心医院眼科,四川遂宁629000

出　　处：《四川医学》2022年第11期1066-1072,共7页Sichuan Medical Journal

基　　金：四川省科技创新苗子工程(编号:2020080)。

摘　　要：目的探讨miR-137靶向Notch1调控对人视网膜微血管内皮细胞(HRCEC)增殖、周期和凋亡的影响。方法使用RT-qPCR检测我院20例高血压视网膜病变患者和20例高血压视网膜正常患者血清miR-137表达水平。将miR-137阴性对照(miR-NC)和miR-137过表达组(mimics)转染至HRCEC中,转染24 h后RT-qPCR检测miR-137和Notch1的RNA表达水平,CCK-8法检测细胞增殖,流式细胞术检测细胞周期和凋亡。运用生物信息学预测和荧光素酶报告实验验证miR-137与Notch1存在靶向性。Western Blot检测Notch1以及与周期和凋亡相关蛋白的表达。结果RT-qPCR结果显示,高血压视网膜病变患者血清中的miR-137表达量显著高于高血压视网膜正常组患者(P<0.05)。细胞增殖实验结果表明,miR-137mimics组细胞增殖指数显著低于miR-NC组(P<0.05)。转染后24 h,miR-137mimics组过表达组的G0/G1期百分比(65.00±1.29)%相对于miR-NC组(56.52±0.81)%显著增加(P<0.05),G2期百分比(17.58±0.75)%则显著低于对照组(22.29±1.57)%(P<0.05);miR-137mimics组的凋亡率(12.67±0.31)%显著高于miR-NC组(2.16±0.12)%(P<0.05)。生物信息学方法预测到miR-137与Notch1存在靶向性。荧光素酶报告实验说明miR-137可以靶向结合Notch1。miR-137mimics组的Notch1 mRNA和蛋白的相对表达量均显著低于miR-NC组(P<0.05),miR-137mimics组的周期相关蛋白CDK2相比miR-NC组表达量显著下降(P<0.05),miR-137mimics组抗凋亡蛋白Bcl-2的表达量相比miR-NC组显著降低(P<0.05),而miR-137mimics组的促凋亡蛋Bax表达量相比miR-NC组则显著增加(P<0.05)。结论miR137能抑制Notch1信号通路,抑制细胞增殖和周期,促进细胞凋亡,提示miR-137可作为预测高血压视网膜病变的潜在分子标志物。Objective To investigate the effects of miR-137 targeting Notch1 on proliferation,cycle and apoptosis of human retinal microvascular endothelial cells.Methods The RT-qPCR was used to measure miR-137 in serum of 20 patients with hypertensive retinopathy and 20 patients with normal retina.The miR-137 negative control(miR-NC)group and miR-137 mimics group(mimics)were transfected into human retinal microvascular endothelial cells(HRCEC).After 24 hours of transfection,RNA expression levels of miR-137 and Notch1 were detected by RT-qPCR.Cell proliferation was detected by CCK-8 method.Cell cycle and apoptosis were detected by flow cytometry.Notch1 protein and expression of cycle and apoptosis related proteins were detected by Western Blot.Results Expression of miR-137 in patients with hypertensive retinopathy was significantly higher than that in patients with hypertensive normal retina(P<0.05).Results of cell proliferation experiment showed that proliferation index of miR-137 mimics group was significantly lower than that of miR-NC group(P<0.05).24 hours after transfection,percentage of G0/G1 phase in miR-137 mimics group(65±1.29)%was significantly higher than that in miR-NC group(56.52±0.81)%(P<0.05),percentage of G2 phase in miR-137 mimics group(17.58±0.75)%was significantly lower than that in control group(22.29±1.57)%(P<0.05),and apoptosis rate in miR-137 overexpression group(12.67±0.31)%was significantly higher than that in miR-NC group(2.16±0.12)%(P<0.05).Bioinformatics methods predicted that miR-137 and Notch1 were targeted.Luciferase reporter assay showed that miR-137 could target Notch1.Relative expression of Notch1 mRNA and protein in miR-137 overexpression group was significantly lower than that in miR-NC group(P<0.05).The expression of cycle related protein CDK2 in miR-137 overexpression group was significantly lower than that in miR-NC group(P<0.05).There was no significant difference in anti apoptotic protein Bcl-2 between miR-137 overexpression group and miR-NC group(P>0.05).However,expression of B

关 键 词：高血压视网膜病变 人视网膜微血管内皮细胞 miR-137 NOTCH1 

分 类 号：R774.1[医药卫生—眼科]