The Effect Of MiR-193a-3p,miR-200a-3p,miR-141-3p On TGF-?2 Gene Expression In H9C2 Rat Cardiomyocytes

Objective : Transforming growth factor-?(TGF-?)signaling pathway is closely related to heart development and cardiovascular disease.MicroRNAs(miRNAs)can reduce the stability of the messenger RNA(mRNA)or inhibit the translation of the mRNA by targeting its 3'-untranslatedregion(3'-UTR),there by participate in the regulation of gene post-transcription.The role and pathogenesis of miRNAs in the regulation of TGF-? signaling pathway in cardiovascular development and disease are unknown.In this study,we investigate the effect of miR-193a-3p,miR-200a-3p,miR-141-3p on the expression of TGF-?2 to explore whether miR-193a-3p,miR-200a-3p,miR-141-3p downregulate the expression of TGF-?2 in H9C2 rat cardiomyocytes.Method: 1.Bioinformatics methods were used to predict the miRNAs,which may affect the TGF-?2 gene expression.2.Wild type and mutant type luciferase reports of TGF-?2 3'-UTR were constructed respectively,and luciferase assays were performed after co-transfected with miR-193a-3p mimics,miR-200a-3p mimics,miR-141-3p mimics respectively.3.miR-193a-3p mimics,miR-200a-3p mimics,miR-141-3p mimics was transfected into the H9C2 rat cardiomyocytes.RT-PCR was used to detect the expression levels of miRNAs and TGF-?2 mRNA.Western blot was used to detect the expression levels of TGF-?2 protein.Result: 1.The results of prediction by Bioinformatics methods showed that there may be targeted relationship between the TGF-?2 gene and miR-193a-3p,miR-200a-3p,miR-141-3p.2.miR-193a-3p mimics could decrease the luciferase activity of wild type TGF-?2 3'-UTR(P<0.05),but had no significantly effect on the luciferase activity of mutant type TGF-?2 3'-UTR(P > 0.05).miR-200a-3p mimics,miR-141-3p mimics had no significantly effect on wither wild type TGF-?2 3'-UTR or mutant type TGF-?2 3'-UTR(P>0.05).3.Compared with negative control,miR-193a-3p,miR-200a-3p,miR-141-3p expression ware significantly increased(P < 0.05)after transfected with miR-193a-3p mimics,miR-200a-3p mimics,miR-141-3p mimics respectively,but the expression of TGF-?2 mRNA and TGF-?2 protein expression were decreased(P<0.05).Conclusion:1.miR-193a-3p can direct target down-regulate the expression of TGF-?2 in H9C2 rat cardiomyocytes.2.miR-200a-3p and miR-141-3p can down-regulate the expression of TGF-?2 in H9C2 rat cardiomyocytes.