MicroRNA-574-5p对冠状动脉平滑肌细胞生长的影响及其调控机制

MicroRNA-574-5p promotes cell growth of smooth muscle cells of coronary artery and its regulative molecular mechanism

目的:研究miRNA-574-5p对冠状动脉平滑肌细胞(Vascular smooth muscle cells,VSMCs)增殖的影响及其调控的分子机制。方法:实时定量PCR检测冠心病(coronary artery heart disease,CAD)患者及健康体检者外周血中miRNA-574-5p的表达及正常人VSMCs细胞中miRNA-574-5p及ZDHHC14的表达。MTT法检测细胞增殖,流式细胞仪检测细胞凋亡。双荧光素酶报告分析检测miRNA-574-5p与靶基因的结合。结果:与正常体检者对比,CAD患者血清中miRNA-574-5p的表达显著增加(P<0.05)。与阴性对照组相比,miRNA-574-5p类似物显著提高VSMCs的增殖(P<0.01),降低VSMCs的凋亡(P<0.01)。经在线miRNA靶基因预测软件分析,miRNA-574-5p的一个靶基因为ZDHHC14。miRNA-574-5p降低ZDHHC14 3'UTR端的荧光素酶活性(P<0.01),且抑制其表达(P<0.01)。ZDHHC14过表达降低miRNA-574-5p诱导的VSMCs的增殖(P<0.05),提高miRNA-574-5p诱导的VSMCs的凋亡(P<0.05)。结论:miR-574-5p通过抑制ZDHHC14表达促进VSMCs增殖,并抑制其凋亡。miR-574-5p可能为CAD相关因子,并可能成为CAD治疗的潜在分子靶点。

Objective:To investigate the effect and regulative molecular mechanism of vascular miRNA-574 -5p on cell proliferation of smooth muscle cells (VSMCs) of coronary artery. Methods:Real-time quantitative PCR was used to detect expression of miRNA-574-5p in peripheral blood of patients with coronary heart disease and healthy people,and to measure expressions of miRNA-574-5p and ZDHHC14 in normal VSMCs.Cell proliferation was detected by MTT method.Apoptosis was tested by flow cytometry.miRNA-574-5p binding to target genes was measured by dual-luciferase reporter assay. Results:Compared with healthy people,expressions of miRNA-574-5p in serum of CAD patients were significantly increased ( P < 0.05).Compared with negative control group,miRNA-574-5p mimic significantly promoted proliferation of VSMCs ( P <0.01),reduced apoptosis of VSMCs ( P <0.01).Online miRNA target gene prediction software analysis shown that ZDHHC14 was one of the target genes of miRNA-574-5p.miRNA-574-5p reduced luciferase activity of ZDHHC14 3 UTR ( P <0.01),and inhibited the expression of ZDHHC14 ( P <0.01).ZDHHC14 overexpression inhibited miRNA-574-5p-induced proliferation of VSMCs ( P <0.05),promoted miRNA-574-5p-induced apoptosis of VSMCs ( P <0.05). Conclusion:miRNA-574-5p promotes proliferation of VSMCs and suppresses apoptosis of VSMCs by inhibiting ZDHHC14 expression,suggesting that miR-574-5p is a CAD-related factor that may serve as a potential molecular target for CAD treatment.