title: Ossotide promotes cell differentiation of human osteoblasts from osteogenesis imperfecta patients by up-regulating miR-145.
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, 200μg/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.
authors.affiliation: Department of Pediatric Orthopedics, Zhengzhou Children's Hospital, Zhengzhou, Henan 450000, China.
authors.forename: Keming
authors.initials: K
authors.lastname: Sun
authors.affiliation: Department of Pediatric Orthopedics, Zhengzhou Children's Hospital, Zhengzhou, Henan 450000, China.
authors.forename: Junjian
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Pediatric Orthopedics, Zhengzhou Children's Hospital, Zhengzhou, Henan 450000, China.
authors.forename: Fangna
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Pediatric Orthopedics, Zhengzhou Children's Hospital, Zhengzhou, Henan 450000, China.
authors.forename: Zejuan
authors.initials: Z
authors.lastname: Ji
authors.affiliation: Department of Pediatric Orthopedics, Zhengzhou Children's Hospital, Zhengzhou, Henan 450000, China.
authors.forename: Zhanhao
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Department of Pediatric Orthopedics, Zhengzhou Children's Hospital, Zhengzhou, Henan 450000, China.
authors.forename: Chunxu
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Pediatric Orthopedics, Zhengzhou Children's Hospital, Zhengzhou, Henan 450000, China. Electronic address: manyeyaoe@yeah.net.
authors.forename: Manye
authors.initials: M
authors.lastname: Yao
mesh_terms: D016022:Case-Control Studies; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D045424:Complex Mixtures / Q000494:pharmacology*; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D018450:Disease Progression; D015536:Down-Regulation / Q000187:drug effects; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010013:Osteogenesis Imperfecta / Q000235:genetics* / Q000473:pathology*; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics*; D060449:Wnt Signaling Pathway / Q000187:drug effects
chemical_list: D045424:Complex Mixtures; D050659:Core Binding Factor Alpha 1 Subunit; C541010:MIRN145 microRNA, human; D035683:MicroRNAs
keywords: Differentiation; MiR-145; Ossotide; Osteoblasts; Runx2