长链非编码RNA M通过作为miRNA-1的中心诱导血管平滑肌细胞增殖和迁移

LncRNA M induces proliferation and migration of vascular smooth muscle cells by acting as a center of miRNA-1

目的:探究LncRNA M是否可以通过作为miRNA-1的中心来诱导血管平滑肌细胞(VSMC)增殖及迁移。方法:将体外培养的VSMC按照各检测指标需要进行转染,通过生物信息学找到可以与LncRNA M结合的miRNA-1;通过双荧光素酶报告基因检测各组的荧光素酶活性;RT-PCR检测LncRNA M与miRNA-1之间的调控关系,并用流式细胞仪检测细胞周期变化,从而找出LncRNA M及miRNA-1对细胞增殖的影响;通过MTT实验和Transwell实验分别检测细胞的增殖、迁移情况。结果:通过生物信息学及双荧光素酶报告基因检测得出LncRNA M可能是miRNA-1的一个靶向调控基因,并且miRNA-1能够和LncRNA M结合从而调控其在VSMC中的表达;RT-PCR和流式细胞术研究发现,敲低LncRNA M可能通过上调miRNA-1的表达来促进VSMC细胞的增殖。通过MTT实验和Transwell实验发现,与对照组相比miRNA-1 mimics组、miRNA-1 mimics+pmirGLO组的细胞活力和迁移率明显上升(P<0.05),过表达LncRNA M后其细胞活力和细胞迁移率明显下降(P<0.05)。结论:LncRNA M可能通过作为miRNA-1的中心上调miRNA-1的表达诱导VSMC的增殖和迁移。

Objective:To explore whether LncRNA M can induce the proliferation and migration of vascular smooth muscle cells(VSMC)by acting as the center of miRNA-1.Methods:VSMC cultured in vitro were transfected according to the requirements of each detection index. miRNA-1 binding to LncRNA M was found by bioinformatics. Luciferase activity was detected by double luciferase reporter gene. Regulatory relationship between LncRNA M and miRNA-1 was detected by RT-PCR,Flow cytometry was used to detect cell cycle changes,so as to find out the relationship between LncRNA M and miRNA-1 and cell proliferation;MTT and Transwell experiments were used to detect cell proliferation and migration.Results:Bioinformatics and double luciferase reporter gene detection showed that LncRNA M might be a target regulatory gene of miRNA-1,and that miRNA-1 could bind with LncRNA M to regulate its expression in VSMC. In order to explore the regulatory relationship between LncRNA M and miRNA-1,RT-PCR and flow cytometry was found that knocking down LncRNA M may promote the proliferation of VSMC by up-regulating the expression of miRNA-1.Through MTT and Transwell experiments,we found that compared with the control group,the cell viability and mobility of the miRNAs-1 mimics group and the microRNAs-1 mimics+pmirGLO group increased significantly(P<0.05),while the cell viability and cell mobility decreased significantly after overexpression of LncRNA M(P<0.05).Conclusion:LncRNA M may induce VSMC proliferation and migration by up-regulating the expression of miRNA-1 as the center of miRNA-1.