微小RNA-610对血小板源生长因子诱导的血管平滑肌细胞影响的分子机制研究  

Molecular mechanism of microRNA-610 on platelet-derived growth factor-induced vascular smooth muscle cells

作　　者：周健华 卢莎 吕永楠[3] ZHOU Jian-hua;LU Sha;LU Yong-nan(Department of Cardiology,Hubei Provincial Traditional Chinese Medical Hospital,Wuhan 430061,Hubei Province,China;Department of Acupuncture and Moxibustion,Huang Jia Hu Hospital of Hubei University of Traditional Chinese Medicine,Wuhan 430065,Hubei Province,China;Department of Cardiology,People’s Hospital of Wuhan University,Wuhan 430060,Hubei Province,China)

机构地区：[1]湖北省中医院心血管内科,湖北武汉430061 [2]湖北中医药大学黄家湖医院针灸科,湖北武汉430065 [3]武汉大学人民医院心血管内科,湖北武汉430060

出　　处：《中国临床药理学杂志》2020年第15期2294-2298,共5页The Chinese Journal of Clinical Pharmacology

基　　金：湖北省自然科学基金资助项目(ZRMS2017000913)。

摘　　要：目的探讨微小RNA-610(miR-610)对血小板源生长因子-BB(PDGF-BB)诱导的血管平滑肌细胞(VSMCs)的增殖、迁移和侵袭的影响及其机制。方法双荧光素酶报告基因分析法验证miR-610对起始识别复合物1(ORC1)的靶向作用。将VSMCs分为8组:空白组、实验组(PDGF-BB处理)和第1至第6转染组(分别转染miR-NC、miR-610模拟物、si-NC、si-ORC1、miR-610+pcDNA、miR-610+pcDNA-ORC1后,进行PDGF-BB处理)。用噻唑蓝法、Transwell实验分别检测细胞活性和迁移、侵袭能力。结果miR-610靶向负性调控ORC1表达。空白组、实验组、第1转染组至第6转染组细胞活力(48 h)分别为0.41±0.04,0.64±0.07,0.72±0.07,0.48±0.05,0.65±0.06,0.48±0.05,0.45±0.05和0.64±0.06;这8组的迁移细胞数分别为(38.46±6.38),(86.39±8.16),(89.34±8.25),(52.39±5.11),(86.73±8.23),(59.63±5.25),(47.89±4.25)和(71.46±7.03)个;这8组的侵袭细胞数分别为(29.36±5.67),(78.69±7.26),(81.43±8.18),(41.59±6.28),(75.36±7.29),(48.63±4.27),(35.41±5.74)和(62.43±6.52)个。空白组与实验组比较,或第1转染组与第2转染组比较,或第3转染组与第4转染组比较,或第5转染组与第6转染组比较,上述指标的差异均有统计学意义(均P<0.05)。结论miR-610通过下调ORC1抑制PDGF-BB诱导的VSMCs的增殖、迁移和侵袭。Objective To explore the effects of microRNA-610(miR-610)on vascular smooth muscle cells(VSMCs)proliferation,migration and invasion induced by platelet-derived growth factor-BB(PDGF-BB)and the mechanism involved.Methods Double luciferase reporter gene analysis was used to verify the targeting effect of miR-610 on origin recognition complex 1(ORC1).VSMCs were divided into 8 groups:blank group,experimental group(PDGF-BB treatment)and transfection-1,-2,-3,-4,-5,-6 groups(PDGF-BB treatment was performed after transfection of miR-NC,miR-610 simulators,si-NC,si-ORC1,miR-610+pcDNA,and miR-610+pcDNA-ORC1,respectively).Thiazole blue and Transwell assays were used to detect cell viability and migration-invasion ability,respectively.Results Double luciferase reporter gene analysis verified the targeting effect of miR-610 on ORC1.ORC1 targeted and negatively regulated ORC1 expression.The cell viability(48 h)in blank group,experimental group,and transfection-1,-2,-3,-4,-5,-6 groups were 0.41±0.04,0.64±0.07,0.72±0.07,0.48±0.05,0.65±0.06,0.48±0.05,0.45±0.05 and 0.64±0.06;the number of migrating cells in the 8 groups were(38.46±6.38),(86.39±8.16),(89.34±8.25),(52.39±5.11),(86.73±8.23),(59.63±5.25),(47.89±4.25)and(71.46±7.03);the number of invasive cells in the 8 groups were(29.36±5.67),(78.69±7.26),(81.43±8.18),(41.59±6.28),(75.36±7.29),(48.63±4.27),(35.41±5.74)and(62.43±6.52).Compared between experimental group and blank group,or the transfection-2 and transfection-1group,or the transfection-4 and transfection-3 group,or transfection-6 and transfection-5 group,the differences of the above indexes were statistically significant(all P<0.05).Conclusion miR-610 inhibits PDGF-BB induced proliferation,migration and invasion of VSMCs by down-regulating ORC1.

关 键 词：血管平滑肌细胞 增殖 迁移 侵袭 微小RNA-610 起始识别复合物基因1 

分 类 号：R97[医药卫生—药品]