MiR-467g suppresses the osteogenic differentiation and mineralization of mouse preosteoblasts via down-regulation of Runx-2 expression / 中国组织工程研究

ABSTRACT

BACKGROUND: Preliminary studies have found miR-467g inhibits bone regeneration, however, there is little information about the underlying mechanism. OBJECTIVE: To explore the effect of miR-467g on osteogenic differentiation and mineralization of mouse preosteoblasts and the underlying mechanism. METHODS: C57 mouse preosteoblasts were isolated and cultured in vitro. The expression levels of Runx-2, Osterix, and Osteocalcin, as well as alkaline phosphatase and mineralization activities were determined by western blot, real-time PCR, alkaline phosphatase and Alizarin red staining, respectively. miR-467g-overexpressed preosteoblasts were constructed to investigate the effect of miR-467g on osteogenic differentiation and mineralization of preosteoblasts by lipofection transfection. Dual luciferase reporter assay was used to identify whether the 3’UTR of Runx-2 mRNA was a binding target of miR-467g. RESULTS AND CONCLUSION: The primary mouse preosteoblasts had a good osteogenic proliferation and differentiation ability in vitro. Expression level of miR-467g was decreased with the increase in osteogenic induction time. MiR-467g suppressed the osteogenic differentiation and mineralization of mouse preosteoblasts. Luciferase assay confirmed that miR-467g targeted Runx-2 directly. In summary, miR-467g can suppress the osteogenic differentiation and mineralization of mouse preosteoblasts via down-regulation of Runx-2 expression.