title: MicroRNA-10a Influences Osteoblast Differentiation and Angiogenesis by Regulating β-Catenin Expression.
abstract: OBJECTIVE
  Elucidation of the molecular mechanisms governing osteoblast differentiation and angiogenesis are of great importance for improving the treatment of bone-related diseases. In this study, we examined the role of microRNA (miR)-10a in the differentiation of MC3T3-E1 cells and pro angiogenic activity of mouse umbilical vein endothelial cells (MUVECs).
  
  
  METHODS
  The murine pre-osteoblast cell line MC3T3-E1 and MUVECs were used in the experiment. After transfected with miR-10a mimics or inhibitors, with or without LiCl pretreatment, the miR-10a, ALP, Runx2, Osx, OC and Dlx5 expression were assessed by RT-PCR. MC3T3-E1 cells were cultured with BMP2 to differentiate into bone cells, osteogenic differentiation of MC3T3-E1 cells were detected by ALP and ARS staining. Cell viability were analyzed by MTT and the protein expression of β-catenin, LEF1, cyclinD1, MMP2, and VEGF were detected by Western blotting; VEGF and VE-cadherin release were assessed by ELISA, and the migration of MUVECs, as well as tube formation were also detected.
  
  
  RESULTS
  MiR-10a expression was obviously down-regulated during osteogenic differentiation. Overexpression of miR-10a inhibited osteogenic differentiation of MC3T3-E1 cells, effectively decreasing MUVECs proliferation, migration, VEGF expression, VE-cadherin concentrations, and tube formation in vitro, whereas miR-10a silence enhanced those processes. Further mechanism assays demonstrated that overexpression of miR-10a reduced the β-catenin at both protein and transcription level, while pretreatment with Wnt signaling activator Licl partially attenuated the suppression effects of miR-10a overexpression on osteoblast differentiation and angiogenesis.
  
  
  CONCLUSIONS
  Our findings imply that miR-10a plays a suppressive role in osteoblast differentiation of MC3T3-E1 cells and pro angiogenic activity of MUVECs by regulating the β-catenin expression, representing a novel and potential therapeutic target for the treatment of bone regeneration-related diseases.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Zhang
authors.forename: Qingxia
authors.initials: Q
authors.lastname: Zhao
authors.forename: Jianghua
authors.initials: J
authors.lastname: Wang
authors.forename: Xijing
authors.initials: X
authors.lastname: He
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D015536:Down-Regulation; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000502:physiology*; D018919:Neovascularization, Physiologic / Q000235:genetics*; D010006:Osteoblasts / Q000166:cytology*; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism*
chemical_list: C569259:MIRN10 microRNA, mouse; D035683:MicroRNAs; D051176:beta Catenin