Effect Of MiR-27a-3p On Osteoblast Differentiation Of Human Dental Pulp Stem Cells By Targeting YAP1

Objective To explore the function of miR-27a-3p on osteoblast differentiation of h DPSCs by targeting YAP1.In order to find an ideal microenvironment for the directional osteogenic differentiation of dental pulp stem cells and improve the value of h DPSCs as seed cells of bone tissue engineering,and provide new therapeutic targets for clinical bone tissue defects.Methods h DPSCs were isolated and culture.miR-27a-3p mimics,miR-27a-3p inhibitor and miR-27a-3p scramble were transfected into h DPSCs to detect the effects of miR-27a-3p expression levels on osteoblast differentiation of h DPSCs,bone-related relative gene expression levels were detected by real-time PCR(q RT-PCR)and Western blot.The target gene of miR-27a-3p was Yes-related protein 1(YAP1),which was predicted by bioinformatics prediction and verified by dual luciferase reporter gene experiments.The expression of YAP1 protein was analyzed by Western blot after regulating miR-27a-3p expression.3'UTR of YAP1 was cloned and ligated to construct a dual luciferase reporter vector,and then the dual luciferase reporter assay was applied to verify the target of miR-27a-3p,the expression vector pc DNA3.1-YAP1 was constructed,the interaction of miR-27a-3p and YAP1 was analyzed.The effect of overexpression of miR-27a-3p or YAP1 on the proliferation of h DPSCs were analyzed by MTT assay.Results Overexpression of miR-27a-3p could promote osteoblast differentiation of h DPSCs.Bioinformatics analysis indicated that the YAP1 was a target gene of miR-27a-3p.Western blot indicated that miR-27a-3p overexpression down-regulated the expression of YAP1.miR-27a-3p could act at the 3-UTR of YAP1 gene to regulate its expression was showed Dual luciferase reporter assay.Overexpression of YAP1 could reverse the inhibitory effect of miR-27a-3p on proliferation of h DPSCs.Conclusion miR-27a-3p promoted osteogenic differentiation of h DPSCs via downregulating the expression of YAP1.