title: MicroRNA-204 regulates vascular smooth muscle cell calcification in vitro and in vivo.
abstract: OBJECTIVE
  Medial artery calcification is a common macroangiopathy that initiates from a cell-regulated process similar to osteogenesis. Although the mechanisms governing this process remain unclear, epigenomic regulation by specific microRNAs might play a role in vascular smooth muscle cell (VSMC) calcification. In this study, we aimed to investigate whether miR-204 participates in the regulation of VSMC calcification.
  
  
  RESULTS
  We found that miR-204 was suppressed in mouse aortic VSMCs during β-glycerophosphate-induced calcification, whereas Runx2 protein levels were elevated. Overexpression of miR-204 by transfection of miR-204 mimics decreased Runx2 protein levels and alleviated β-glycerophosphate-induced osteoblastic differentiation of VSMCs, whereas miR-204 inhibition by transfection of miR-204 inhibitors significantly elevated Runx2 protein levels and enhanced osteoblastic differentiation of VSMCs, suggesting the role of miR-204 as an endogenous attenuator of Runx2 in VSMC calcification. Luciferase reporter assays revealed Runx2 as the direct target of miR-204 by overexpression of miR-204 on the wild-type or mutant 3'-UTR sequences of Runx2 in VSMCs. In vivo overexpression of miR-204 by injection of miR-204 agomirs in Kunming mice attenuated vitamin D3-induced medial artery calcification.
  
  
  CONCLUSIONS
  Our study has shown that down-regulation of miR-204 may contribute to β-glycerophosphate-induced VSMC calcification through regulating Runx2. miR-204 represents an important new regulator of VSMC calcification and a potential therapeutic target in medial artery calcification.
authors.affiliation: Department of Pediatrics, The Second Xiang-Ya Hospital,  Central South University, #139 Middle Renmin Road, 410011, Changsha, Hunan,  People's Republic of China.
authors.forename: Rong-Rong
authors.initials: RR
authors.lastname: Cui
authors.forename: Shi-Jun
authors.initials: SJ
authors.lastname: Li
authors.forename: Ling-Juan
authors.initials: LJ
authors.lastname: Liu
authors.forename: Lu
authors.initials: L
authors.lastname: Yi
authors.forename: Qiu-Hua
authors.initials: QH
authors.lastname: Liang
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhu
authors.forename: Guan-Ying
authors.initials: GY
authors.lastname: Liu
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
authors.forename: Shan-Shan
authors.initials: SS
authors.lastname: Wu
authors.forename: Xiao-Bo
authors.initials: XB
authors.lastname: Liao
authors.forename: Ling-Qing
authors.initials: LQ
authors.lastname: Yuan
authors.forename: Ding-An
authors.initials: DA
authors.lastname: Mao
authors.forename: Er-Yuan
authors.initials: EY
authors.lastname: Liao
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism*; D005260:Female; D005994:Glycerophosphates / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D010006:Osteoblasts / Q000378:metabolism; D010012:Osteogenesis; D015854:Up-Regulation; D061205:Vascular Calcification / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D050659:Core Binding Factor Alpha 1 Subunit; D005994:Glycerophosphates; C574692:MIRN204 microRNA, mouse; D035683:MicroRNAs; C493716:Runx2 protein, mouse; C031463:beta-glycerophosphoric acid