Study of microRNAs targeted Dvl2 on the osteoblasts differentiation of rat BMSCs in hyperlipidemia environment

Dishevelled 2 (Dvl‐2), a key mediator of the wnt/β‐catenin signaling pathway, plays critical roles in osteoblasts differentiation in hyperlipidemia environment. In our previous study, we observed a strong correlation between increased dvl2 expression and decreased new bone formation around implants in a rat hyperlipidemia implant surgery model. However, transcriptional regulation of Dvl2 by microRNAs in this process remains unknown. In the current study, we searched in online database and identified four significantly up‐regulated miRNAs, miR‐21‐5p, miR‐29c‐3p, miR‐138‐5p, and miR‐351‐5p that could potentially regulate Dvl2. Using Western blot and dual‐luciferase assays, we confirmed that miR29c‐3p suppresses Dvl2 expression by binding to its 3′‐UTR. Our results suggest a novel transcriptional regulation mechanism of Dvl2 by miR‐29c‐3p in osteoblasts differentiation of BMSCs. The expression of Dvl2 protein was visualized by Western blot analysis at 48 hrs after transfected mimic and inhibitor of these four miRNAs. We demostrate that in the high lipid environment, the differentiation of BMSCs to osteoblasts is suppressed, which correlated with down‐regulation of key osteogenesis genes RUNX2, ALP SP7, and Dvl2. We further demonstrated that, among the four predicted miRNAs, miR‐29c‐3p can suppressor the expression of Dvl2. We demonstrated in this research that miR‐29c‐3p suppresses Dvl2 expression by specific binding to its 3′‐UTR promoter region.