Ossotide promotes cell differentiation of human osteoblasts from osteogenesis imperfecta patients by up-regulating miR-145

• Published in: Biomedicine & pharmacotherapy 2016-10, Vol.83, p.1105-1110
• Main Authors: Sun, Keming, Wang, Junjian, Liu, Fangna, Ji, Zejuan, Guo, Zhanhao, Zhang, Chunxu, Yao, Manye
• Format: Article
• Language: English
• Subjects: Case-Control Studies; Cell Differentiation - drug effects; Cell Proliferation - drug effects; Complex Mixtures - pharmacology; Core Binding Factor Alpha 1 Subunit - genetics; Core Binding Factor Alpha 1 Subunit - metabolism; Differentiation; Disease Progression; Down-Regulation - drug effects; Humans; Internal Medicine; Medical Education; MicroRNAs - genetics; MicroRNAs - metabolism; MiR-145; Ossotide; Osteoblasts; Osteoblasts - drug effects; Osteoblasts - metabolism; Osteoblasts - pathology; Osteogenesis Imperfecta - genetics; Osteogenesis Imperfecta - pathology; Runx2; Up-Regulation - drug effects; Up-Regulation - genetics; Wnt Signaling Pathway - drug effects
• ISSN: 0753-3322; 1950-6007
• DOI: 10.1016/j.biopha.2016.08.025

Abstract Ossotide as an effective bone formation compound preparation has been proved to promote osteoblasts differentiation. MiR-145 is significantly decreased in osteogenesis imperfecta (OI) patients, but it is still unknown whether ossotide performed its effect by regulating miR-145. In this study, we investigated the effect of ossotide on regulating miR-145 expression and osteoblasts differentiation. The primary osteoblasts cells were isolated from OI patients and then cultured with different concentrations (0, 25, 50, 100, 200 μg/l) of ossotide. The cell proliferation was detected with CCK-8 Elisa kit after ossotide treatment. The level of miR-145 expression was determined using qRT-PCR. In order to study whether ossotide up regulated miR-145, miR-145 mimic and miR-145 inhibitor were used to up regulate and down regulate the miR-145 levels in osteoblasts. The expressions of Runx2, Osx, β-catenin, TCF-1 were detected using Western blot and qRT-PCR. We observed that miR-145 was up regulated by ossotide treatment in miR-145 mimic or miR-145 inhibitor treated osteoblasts. What’s more, up regulated miR-145 increased the expression of osteoblasts differentiation regulated protein Runx2 and Osx. In addition, Wnt signaling related β-catenin, TCF-1 were activated by up-regulated miR-145 which was induced by ossotide treatment. In summary, ossotide induced cell differentiation and Wnt signaling activation in osteoblasts by up regulating miR-145.