Background:microRNAs are crucial for cardiovascular development and are associated with congenital heart disease(CHD).Recent studies have shown that microRNAs play a role in heart development and is closely related to...Background:microRNAs are crucial for cardiovascular development and are associated with congenital heart disease(CHD).Recent studies have shown that microRNAs play a role in heart development and is closely related to CHD.The present study investigated the underlying mechanism of microRNA-208a(miR-208a)in“simple”CHD.Material and Methods:Reverse transcription-quantitative PCR(RT-qPCR)demonstrated miR-208a expression levels in children with CHD(n=27)compared with normal controls(n=29),in cardiomyocytes from embryo 10(E10)to post-birth(P7)and organs in adult rats in healthy rats.Apoptosis of H9c2 cells after transfection with miR-208a detected by TUNEL assay.B-cell lymphoma(Bcl)-2,an anti-apoptotic gene,was detected by RT-qPCR,as well as Gata4.After 48h overexpression of miR-208a,GATA4 was detected via western blotting.Dual luciferase reporting system was used to identify the binding sites of miR-208a to Gata4.Results:Expression of miR-208a was upregulated in the CHD group via the control group(p<0.01).At P7,miR-208a had the highest expression(p<0.01),and which was highest in myocardiocytes via other organs or tissues(p<0.01)in adult rats.The number of apoptotic cells increased significantly post-transfection with miR-208a(p<0.01),while decreased with the miR-208a inhibitor via the control group(p<0.01).Compared with the control group,there was no significant difference in the expression level of Bcl-2 after miR-208a overexpression(p>0.05).The present study proved that miR-208a binds directly to the 3´-UTR of Gata4 at site 1,363-1,369 bp.Expression of GATA4 decreased after miR-208a overexpression(p<0.01),but increased following transfection with a miR-208a inhibitor via the control group(p<0.05).Conclusions:Our study demonstrated that miR-208a downregulates Bcl-2 by directly targeting GATA4,which may cause CHD.miR-208a may become a new biomarker or therapeutic target for CHD in the future.展开更多
基金The work was supported by the National Science Foundation of China(No.8170131).
文摘Background:microRNAs are crucial for cardiovascular development and are associated with congenital heart disease(CHD).Recent studies have shown that microRNAs play a role in heart development and is closely related to CHD.The present study investigated the underlying mechanism of microRNA-208a(miR-208a)in“simple”CHD.Material and Methods:Reverse transcription-quantitative PCR(RT-qPCR)demonstrated miR-208a expression levels in children with CHD(n=27)compared with normal controls(n=29),in cardiomyocytes from embryo 10(E10)to post-birth(P7)and organs in adult rats in healthy rats.Apoptosis of H9c2 cells after transfection with miR-208a detected by TUNEL assay.B-cell lymphoma(Bcl)-2,an anti-apoptotic gene,was detected by RT-qPCR,as well as Gata4.After 48h overexpression of miR-208a,GATA4 was detected via western blotting.Dual luciferase reporting system was used to identify the binding sites of miR-208a to Gata4.Results:Expression of miR-208a was upregulated in the CHD group via the control group(p<0.01).At P7,miR-208a had the highest expression(p<0.01),and which was highest in myocardiocytes via other organs or tissues(p<0.01)in adult rats.The number of apoptotic cells increased significantly post-transfection with miR-208a(p<0.01),while decreased with the miR-208a inhibitor via the control group(p<0.01).Compared with the control group,there was no significant difference in the expression level of Bcl-2 after miR-208a overexpression(p>0.05).The present study proved that miR-208a binds directly to the 3´-UTR of Gata4 at site 1,363-1,369 bp.Expression of GATA4 decreased after miR-208a overexpression(p<0.01),but increased following transfection with a miR-208a inhibitor via the control group(p<0.05).Conclusions:Our study demonstrated that miR-208a downregulates Bcl-2 by directly targeting GATA4,which may cause CHD.miR-208a may become a new biomarker or therapeutic target for CHD in the future.
