干扰miR-148a-5p对肿瘤坏死因子-α诱导血管平滑肌细胞增殖和迁移的影响  

Effect of Interfering with miR-148a-5p on Proliferation and Migration of Vascular Smooth Muscle Cells Induced by TNF-α

作　　者：陈愿[1] 赵子明 宋爽[1] 崔留义 李传荣 杨晓航 沈蕾 刘乾坤 CHEN Yuan;ZHAO Ziming;SONG Shuang;CUI Liuyi;LI Chuanrong;YANG Xiaohang;SHEN Lei;LIU Qiankun(The Seventh People′s Hospital of Zhengzhou,Zhengzhou 450001,Henan,China)

机构地区：[1]郑州市第七人民医院,郑州450001

出　　处：《中西医结合心脑血管病杂志》2021年第21期3674-3678,共5页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：河南省医学科技攻关计划项目(No.2018020855)。

摘　　要：目的探讨干扰miR-148a-5p对肿瘤坏死因子-α(TNF-α)诱导的血管平滑肌细胞(VSMC)增殖和迁移的影响。方法体外培养VSMC,分为对照组、TNF-α组、TNF-α+anti-miR-NC组和TNF-α+anti-miR-148a-5p组,实时荧光定量聚合酶链式反应(RT-qPCR)检测细胞中miR-148a-5p表达水平,流式细胞术检测细胞周期和凋亡,蛋白免疫印迹(Western Blot)法检测细胞中细胞核增殖抗原Ki67、上皮型钙黏附蛋白(E-cadherin)和神经型钙黏附蛋白(N-cadherin)表达水平。双荧光素酶报告基因实验验证miR-148a-5p与第10号染色体缺失的磷酸酶张力蛋白同源物基因(PTEN)调控关系。结果与对照组比较,TNF-α组细胞中miR-148a-5p的表达水平升高(P<0.05),G0/G1期明显降低(P<0.05),S期明显升高(P<0.05),细胞迁移数和Ki67、N-cadherin蛋白表达水平明显升高(P<0.05),E-cadherin蛋白表达水平明显降低(P<0.05)。与TNF-α+anti-miR-NC组比较,TNF-α+anti-miR-148a-5p组细胞G0/G1期明显升高(P<0.05),S期明显降低(P<0.05),细胞迁移数和Ki67、N-cadherin蛋白表达水平明显降低(P<0.05),E-cadherin蛋白表达水平明显升高(P<0.05)。双荧光素酶报告基因结果显示,miR-148a-5p可降低PTEN野生型的荧光素酶活性(P<0.05)。结论干扰miR-148a-5p表达可抑制TNF-α诱导的VSMC增殖和迁移。Objective To investigate the effects of interfering with miR-148a-5p on the proliferation and migration of vascular smooth muscle cells(VSMC)induced by tumor necrosis factor-α(TNF-α).Methods VSMCs were cultured in vitro and divided into control group,TNF-αgroup,TNF-α+anti-miR-NC group,and TNF-α+anti-miR-148a-5p group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of miR-148a-5p,flow cytometry was used to detect the cell cycle and apoptosis.Western Blot was used to detect the expression levels of Ki67,E-cadherin and N-cadherin protein.The dual luciferase reporter gene experiment was used to verified the regulatory relationship between miR-148a-5p and phosphatase and tensin homolog deleted on chromosome ten(PTEN).Results Compared with control group,the expression level of miR-148a-5p in TNF-αgroup was significantly increased(P<0.05),the G0/G1 phase was significantly shortened(P<0.05),the S phase was significantly extended(P<0.05),the number of migrated cells and the expression levels of Ki67 and N-cadherin protein were significantly increased(P<0.05),and the expression level of E-cadherin protein was significantly reduced(P<0.05).Compared with TNF-α+anti-miR-NC group,the G0/G1 phase of TNF-α+anti-miR-148a-5p group was significantly prolonged(P<0.05),the S phase was significantly shortened(P<0.05),the number of migrated cells and the expression levels of Ki67 and N-cadherin protein were significantly reduced(P<0.05),and the expression level of E-cadherin protein was significantly increased(P<0.05).The dual luciferase reporter gene results showed that miR-148a-5p could reduce the luciferase activity of PTEN wild type(P<0.05).Conclusion Interference of with the expression of miR-148a-5p can inhibit the proliferation and migration of VSMCs induced by TNF-α.

关 键 词：血管平滑肌细胞 miR-148a-5p 肿瘤坏死因子-Α 细胞增殖 细胞迁移 实验研究 

分 类 号：R73[医药卫生—肿瘤]