MiR-132-3p regulates ADAMTS-5 expression and promotes chondrogenic differentiation of rat mesenchymal stem cells

J Cell Biochem. 2018 Mar;119(3):2579-2587. doi: 10.1002/jcb.26421. Epub 2017 Dec 12.

Abstract

Previous study showed that miRNA aberrant expression is involved in chondrogenic differentiation. In this study, we aimed to investigate the effects of miR-132-3p on chondrogenic differentiation and the underlying mechanisms. First, quantitative PCR were performed to determine the level of MiR-132-3p. Then, we used luciferase assay to examine the target of miR-132-3p. Proteoglycan was tested by Alcian blue staining assay. Moreover, the sex determining region Y-box 9 (SOX9), Collagen type II alpha 1 chain (COL2A1) and Aggrecan (ACAN) levels were analyzed by quantitative PCR, immunofluorescence and Western blotting. Our results showed that MiR-132-3p level was reduced in rat MSCs (rMSCs) during chondrogenic differentiation. Ectopic expression of miR-132-3p induced proteoglycan accumulation and the increase of ACAN, SOX9 and COL2A1 expression, which were involved in inducing chondrogenic differentiation of rMSCs. More importantly, ADAMTS-5 was identified as the target of MiR-132-3p. Knockdown of ADAMTS-5 increased proteoglycan level, but reduced the SOX9, ACAN, and COL2A1 levels during chondrogenic differentiation of rMSCs. Taken together, our results revels that MiR-132-3p promotes rMSCs chondrogenic differentiation, possibly mediated by targeting ADAMTS-5, which provided new perspective on the chondrogenic differentiation and pathology of osteoarthritis.

Keywords: ADAMTS-5; chondrogenic differentiation; miR-132-3p; rat mesenchymal stem cells.

MeSH terms

  • ADAMTS5 Protein / biosynthesis*
  • ADAMTS5 Protein / genetics
  • Animals
  • Cell Differentiation / genetics*
  • Cells, Cultured
  • Chondrocytes / cytology*
  • Chondrogenesis / genetics
  • Gene Expression Regulation / genetics*
  • Male
  • Mesenchymal Stem Cells / cytology*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Rats

Substances

  • MIRN132 microRNA, rat
  • MicroRNAs
  • ADAMTS5 Protein
  • Adamts5 protein, rat