SLIT1 3'UTR Modulates the Phenotype of Endothelial Cells Through MiR-34a-5p/SIRT1 Axis
In the evolutionary process of higher organisms,the 3'untranslated region(3'UTR)sequence of mRNA markedly increase,which indicates a potential important role of 3'UTR in regulating biological functions.The 3'UTR of slit guidance ligand 1(SLIT1)mRNA was markedly decreased in the myocardium of hypertrophic cardiomyopathy(HCM)patients,however,the potential role and mechanism of SLIT 1 3'UTR in regulating vascular function in the hypertrophic myocardium remains unclear.The expression of endothelial nitric oxide synthase(eNOS)and vascular endothelial growth factor A(VEGFA)was detected in aortic endothelial cells(HAECs)with overexpression of SLIT1 3'UTR.The migration and tube formation activities of HAECs were examined by using wound-healing assay and matrigel-based tube formation assay,respectively.Overexpression of SLIT1 3'UTR could significantly increase the levels of p-eNOS,eNOS and VEGFA in HAECs(P＜0.01,respectively),and promote the migration and tube formation activities of HAECs(P＜0.01,respectively).Bioinformatic analysis indicated multiple potential binding sites of microRNAs(miRNAs)exist in SLIT1 3'UTR.The results of RNA antisense purification(RAP)assay,as well as Ago2-based RNA binding protein immunoprecipitation(RIP)assay,verified the interaction between SLIT1 3'UTR and miR-34a-5p.Additionally,a significant reduction of miR-34a-5p was observed in HAECs with overexpression of SLIT13'UTR(P＜0.05),and transfection of miR-34a-5p mimic could reverse the increase of SIRT1 expression by SLIT1 3'UTR in HAECs(P＜0.05).Transfection of miR-34a-5p and SIRT1 siRNA could consistently suppress the increase of p-eNOS,eNOS and VEGFA,and the enhancement of migration and tube formation activities of HAECs with overexpression of SLIT1 3'UTR(P＜0.05,P＜0.01,respectively).In conclusion,SLIT1 3'UTR can sponge miR-34a-5p,and promote the endothelial cell migration and tube formation by targeting miR-34a-5p/SIRT1 axis.
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