MiRNA-20a promotes osteogenic differentiation of human mesenchymal stem cells by co-regulating BMP signaling

RNA Biol. Sep-Oct 2011;8(5):829-38. doi: 10.4161/rna.8.5.16043. Epub 2011 Jul 28.

Abstract

Osteogenic differentiation of mesenchymal stem cells (MSCs) is a complex process, which is regulated by various factors including microRNAs. Our preliminary data showed that the expression of endogenous miR-20a was increased during the course of osteogenic differentiation. Simultaneously, the expression of osteoblast markers and regulators BMP2, BMP4, Runx2, Osx, OCN and OPN was also elevated whereas adipocyte markers PPARγ and osteoblast antagonist, Bambi and Crim1, were downregulated, thereby suggesting that miR-20a plays an important role in regulating osteoblast differentiation. To validate this hypothesis, we tested its effects on osteogenic differentiation by introducing miR-20a mimics and lentiviral-miR20a-expression vectors into hMSCs. We showed that miR-20a promoted osteogenic differentiation by the upregulation of BMP/Runx2 signaling. We performed bioinformatics analysis and predicted that PPARγ, Bambi and Crim1 would be potential targets of miR-20a. PPARγ is a negative regulator of BMP/Runx2 signaling whereas Bambi or Crim1 are antagonists of the BMP pathway. Furthermore, we confirmed that all these molecules were indeed the targets of miR-20a by luciferase reporter, quantitative RT-PCR and western blot assays. Similarly to miR-20a overexpression, the osteogenesis was enhanced by the silence of PPARγ, Bambi or Crim1 by specific siRNAs. Taken together, for the first time, we demonstrated that miR-20a promoted the osteogenesis of hMSCs in a co-regulatory pattern by targeting PPARγ, Bambi and Crim1, the negative regulators of BMP signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Morphogenetic Protein Receptors
  • Bone Morphogenetic Proteins / antagonists & inhibitors
  • Bone Morphogenetic Proteins / metabolism*
  • Cell Differentiation / genetics
  • Cells, Cultured
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Genetic Vectors / genetics
  • Humans
  • Lentivirus / genetics
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mesenchymal Stem Cells / metabolism*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Osteogenesis / genetics*
  • Osteoporosis / genetics
  • Osteoporosis / metabolism
  • PPAR gamma / genetics
  • PPAR gamma / metabolism
  • RNA Interference
  • RNA, Small Interfering
  • Signal Transduction

Substances

  • BAMBI protein, human
  • Bone Morphogenetic Proteins
  • CRIM1 protein, human
  • Core Binding Factor Alpha 1 Subunit
  • MIRN20a microRNA, human
  • Membrane Proteins
  • MicroRNAs
  • PPAR gamma
  • RNA, Small Interfering
  • RUNX2 protein, human
  • Bone Morphogenetic Protein Receptors