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MicroRNA-145 regulates chondrogenic differentiation of mesenchymal stem cells by targeting Sox9

Bo Yang1, Hongfeng Guo, Yulan Zhang

  • 1Laboratory of Biomechanics, Department of Anatomy, The Third Military Medical University, Chongqing, People's Republic of China.

Plos One
|July 30, 2011
PubMed

Insights

MicroRNAs (miRNAs) regulate chondrogenesis. This study found that miR-145, a microRNA, negatively regulates chondrogenic differentiation by targeting the SOX9 (SRY-related high mobility group-box gene 9) transcription factor.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Stem Cell Biology

Background:

  • Mesenchymal stem cell (MSC) chondrogenesis is crucial for cartilage formation and repair.
  • The precise molecular mechanisms, including microRNA (miRNA) involvement, regulating chondrogenesis remain incompletely understood.
  • Transforming growth factor beta 3 (TGF-β3) is a key inducer of chondrogenic differentiation.

Purpose of the Study:

  • To investigate the role of miRNAs in TGF-β3-induced chondrogenic differentiation of MSCs.
  • To identify specific miRNAs and their targets involved in regulating chondrogenesis.
  • To elucidate the regulatory mechanism of miR-145 in chondrogenic differentiation.

Main Methods:

  • Quantitative real-time PCR to measure miRNA and mRNA expression.
  • Dual-luciferase reporter gene assay to validate miRNA-target interaction.
  • Overexpression and inhibition of miR-145 in C3H10T1/2 cells.
  • Western blot analysis for protein expression.
  • Analysis of chondrogenic marker gene expression (Col2a1, Agc1, COMP, Col9a2, Col11a1).

Main Results:

  • miR-145 expression was decreased during TGF-β3-induced chondrogenic differentiation.
  • miR-145 directly targets the 3'-UTR of SOX9 (SRY-related high mobility group-box gene 9) mRNA.
  • Overexpression of miR-145 reduced SOX9 protein levels and chondrogenic marker gene expression.
  • Inhibition of miR-145 increased SOX9 protein levels and chondrogenic marker gene expression.

Conclusions:

  • miR-145 acts as a key negative regulator of chondrogenic differentiation.
  • miR-145 directly targets SOX9, a critical transcription factor for chondrogenesis.
  • This regulatory axis involving miR-145 and SOX9 is important at the early stage of chondrogenesis.

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