miR-144 promotes the proliferation and differentiation of bone mesenchymal stem cells by downregulating the expression of SFRP1

Abstract

Osteoporosis (OP) seriously affects the health and quality of life of elderly individuals and postmenopausal women, and the need to identify drugs that can prevent or treat OP remains urgent. Recently, several miRNAs have been reported to be involved in the differentiation of mesenchymal stem cells and osteoblasts; however, the role of miRNA (miR)-144 in regulating OP remains to be elucidated. In the present study, the expression levels of miR-144, secreted frizzled-related protein 1 (Sfrp1) and TNF-α in clinical samples were detected by the reverse transcription-quantitative polymerase chain reaction analysis and ELISA, respectively. 5-Ethynyl-2′-deoxyuridine staining, Hoechst 33258 staining, flow cytometry, a clone formation assay and Alizarin red staining were used to assess the effects of miR-144 combined with or without Sfrp1 small interfering RNA on the proliferation, apoptosis and osteoblastic differentiation of primary mesenchymal stem cells isolated from rats. Western blot assays were performed to assess the relevant mechanisms, and a dual luciferase reporter assay was used to detect the interaction between miR-144 and Sfrp1. The results showed that the levels of miR-144, Sfrp1 and TNF-α in clinical serum samples obtained from patients with postmenopausal OP were higher than those in serum samples obtained from postmenopausal women with normal bone density. There was a significant positive correlation between miR-144 and Sfrp1. Functional experiments demonstrated that miR-144 promoted proliferation, inhibited apoptosis and induced the osteoblastic differentiation of bone marrow-derived mesenchymal stem cells by targeting Sfrp1. It was also shown that miR-144 may help regulate OP by activating the Wnt/β-catenin pathway. These data suggest miR-144 as a novel target for preventing and treating OP.