title: The influence of MicroRNA-150 in Osteoblast Matrix Mineralization.
abstract: This study investigated the influence of miR-150 expression on osteoblast matrix mineralization and its mechanisms. The mouse osteoblast cell line MC3T3-E1 was used as an in vitro model of bone formation. On the fifth day of mineralization, transfection experiments using agomiR-150, agomiR-NC, antagomiR-150 antagomiR-NC, and mock groups were set up to test the effects of miR-150 in MC3T3-E1 model. The mRNA and protein levels of OC, ALP, type I collagen, and OPN were measured by qRT-PCR and ELISA. Matrix mineralization was detected by alizarin red S (ARS) staining and flow cytometry was employed to quantify apoptosis in each group. RT-PCR and Western blot were applied to detect the expression of target gene MMP14. Our results demonstrated that the endogenous expression levels of miR-150, OC, ALP, type I collagen, and OPN in MC3T3-E1 cells increased steadily. Exogenous expressions of agomiR-150 and antagomiR-150 can significantly up-/down-regulate, respectively, the expression level of miR-150 in MC3T3-E1 cells. Compared with the mock group, higher expression levels of OC, ALP, type I collagen, and OPN mRNA were observed in the agomiR-150 group, while lower mRNA expression levels of OC, ALP, type I collagen, and OPN were found in the antagomiR-150 group. Based on these results, potential miR-150 targeted genes are discussed. Our results showed that miR-150 supports the osteoblastic phenotype related to osteoblast function and bone mineralization. Thus, miR-150 may have potential therapeutic applications in promoting bone formation in certain disease settings, such as in osteoporosis and in elderly patients.
authors.affiliation: Department of Nursing, Linyi People's Hospital, Linyi 276003, P.R. China.
authors.forename: Chun-Ling
authors.initials: CL
authors.lastname: Dong
authors.affiliation: Department of Pharmacology, Linyi Health School, Linyi 276000, P.R. China.
authors.forename: Hao-Zhi
authors.initials: HZ
authors.lastname: Liu
authors.affiliation: Department of Rheumatism Immunity, Linyi People's Hospital, Linyi 276003, P.R. China.
authors.forename: Zhen-Chun
authors.initials: ZC
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Linyi People's Hospital, Linyi 276003, P.R. China.
authors.forename: Huan-Li
authors.initials: HL
authors.lastname: Zhao
authors.affiliation: Department of Rheumatism Immunity, Linyi People's Hospital, Linyi 276003, P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Nursing, Linyi People's Hospital, Linyi 276003, P.R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Nursing, Linyi People's Hospital, Linyi 276003, P.R. China.
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Yao
authors.affiliation: Department of Nursing, Linyi People's Hospital, Linyi 276003, P.R. China.
authors.forename: Tian-Sheng
authors.initials: TS
authors.lastname: Sun
mesh_terms: D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects* / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000378:metabolism; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis*
chemical_list: D035683:MicroRNAs; C529101:Mirn150 microRNA, mouse; D012333:RNA, Messenger
keywords: MICRORNA-150; MINERALIZATION; MIRNA AGOMIR; MIRNA ANTAGOMIR; OSTEOBLAST