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miR-374-5p inhibits osteogenesis by targeting PTEN/PI3K/AKT signaling pathway

Guangning Long1,2,3, Fen Liu1,2,3, Hongmeng Cheng1,2,3

  • 1School of Stomatology, Jiangxi Medical College, Nanchang University, Nanchang, 330006, PR China.

Journal of Orthopaedic Surgery and Research
|March 15, 2025
PubMed
Abstract

Insights

MicroRNA miR-374-5p inhibits bone formation in rat osteoblasts by targeting PTEN and regulating the PI3K/AKT pathway. This study clarifies miR-374-5p

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) are crucial regulators of gene expression.
  • Osteogenesis, the process of bone formation, is tightly controlled by various molecular factors.
  • Dysregulation of miRNAs can contribute to bone-related diseases.

Purpose of the Study:

  • To investigate the role of miR-374-5p in osteogenesis in rat osteoblasts.
  • To identify PTEN as a direct target of miR-374-5p.
  • To elucidate the involvement of the PTEN/PI3K/AKT signaling pathway in miR-374-5p-mediated osteoblast differentiation.

Main Methods:

  • Transfection of cells with miR-374-5p mimics/inhibitors.
  • Dual-luciferase assay to confirm PTEN targeting.
  • qRT-PCR and Western blot to analyze gene and protein expression of osteoblastic markers (Runx2, OSX, OCN) and PTEN.
  • Lentiviral vector for miR-374-5p overexpression in rat osteoblast progenitors.
  • Alizarin Red staining and ALP activity assays for bone formation assessment.
  • Rescue experiments with PI3K/AKT pathway inhibitor (LY294002).

Main Results:

  • miR-374-5p directly targets and downregulates PTEN at both mRNA and protein levels.
  • Overexpression of miR-374-5p suppressed osteoblast differentiation markers (Runx2, OSX, OCN) and reduced bone formation (calcium nodules, ALP activity).
  • Inhibition of miR-374-5p promoted osteogenesis.
  • The PI3K/AKT signaling pathway was modulated by miR-374-5p, and its inhibitory effect on osteogenesis was reversed by LY294002.

Conclusions:

  • miR-374-5p acts as an inhibitor of osteogenesis in rat osteoblasts.
  • The mechanism involves the direct targeting of PTEN and subsequent modulation of the PI3K/AKT signaling pathway.
  • These findings highlight miR-374-5p as a potential therapeutic target for bone-related disorders.
Keywords:
MiRNAOsteogenesisPI3K/AKTPTEN

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