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MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells

Nuo Yin1, Longzhang Zhu1, Liang Ding1

  • 1Department of Orthopaedics, Shanghai Fengxian District Central Hospital, No. 6600, Nanfeng Highway, Shanghai, 201499 China.

Cellular & Molecular Biology Letters
|August 15, 2019
PubMed
Abstract

Insights

MicroRNA-135-5p promotes bone formation by targeting HIF1AN, offering potential therapeutic applications for osteoporosis. This study elucidates the molecular mechanism of miR-135-5p in osteoblast differentiation and calcification.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Background:

  • MicroRNAs (miRNAs) are key regulators in osteoporosis development.
  • Osteoblast differentiation and calcification are critical processes in bone formation.

Purpose of the Study:

  • To investigate the role of miR-135-5p in osteoblast differentiation and calcification.
  • To elucidate the molecular mechanism underlying miR-135-5p's function.

Main Methods:

  • MC3T3-E1 cells were treated with BMP2 and transfected with miR-135-5p mimic or inhibitor.
  • Quantitative RT-PCR, Western blot, ALP activity assay, and alizarin red staining were used to assess osteogenic markers and calcification.
  • Luciferase reporter assays and HIF1AN overexpression were employed to confirm the target and mechanism.

Main Results:

  • Overexpression of miR-135-5p enhanced osteogenic differentiation and calcification, increasing ALP activity and levels of Runx2, OSX, OPN, and OCN.
  • Knockdown of miR-135-5p inhibited these processes.
  • HIF1AN was identified as a direct target of miR-135-5p; its overexpression inhibited osteogenesis, an effect reversed by miR-135-5p.

Conclusions:

  • miR-135-5p promotes osteoblast differentiation and calcification by targeting HIF1AN.
  • miR-135-5p demonstrates potential as a therapeutic agent for enhancing bone formation in conditions like osteoporosis.
Keywords:
CalcificationHypoxia-inducible factor 1 α inhibitorOsteoblast differentiationmiR-135

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