ArticleJournal of orthopaedic surgery and research2025
miR-374-5p inhibits osteogenesis by targeting PTEN/PI3K/AKT signaling pathway.
Article in Journal of orthopaedic surgery and research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- LncRNA-XIST Competitively Binds to MicroRNA-532-5p to Regulate PTEN Expression, Exacerbating Cerebral Ischemia-Reperfusion Injury.Advanced biology · 2026Article
- Inhibition of miR-224-5p promotes osteogenesis in dental pulp stem cells by targeting the PTEN/PI3K/AKT axis.Journal of orthopaedic surgery and research · 2026Article
- Macrophage-derived exosomal miR-466i-5p and miR-365-2-5p stimulated by BMP-2 promote osteogenesis.Journal of orthopaedic surgery and research · 2025Article
- Injectable and Assembled Calcium Sulfate/Magnesium Silicate 3D Scaffold Promotes Bone Repair by In Situ Osteoinduction.Bioengineering (Basel, Switzerland) · 2025Article
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Authors and funding
8 authors.
Funding
Abstract
purposeThis study aims to evaluate the effects of miR-374-5p on osteogenesis in rat osteoblasts, validate its target on PTEN, and explore its role in the PTEN/PI3K/AKT signaling pathway during osteoblast differentiation.
methodsWe transfected 293T cells with miR-374-5p mimics and inhibitors, followed by Western blot and qRT-PCR analyses to assess protein and mRNA expression levels. A dual-luciferase assay was performed to confirm direct targeting. Markers of osteoblast function, such as Runx2, OSX, and OCN, were examined in osteoblasts from rats by qRT-PCR and Western blot. Additionally, we developed a lentiviral vector to overexpress miR-374-5p, which successfully infected rat osteoblast progenitors. Bone formation was subsequently assessed using Alizarin Red staining and ALP activity assays. Finally, rescue experiments were conducted to validate the involvement of the miR-374-5p/PTEN/PI3K/AKT signaling pathway.
resultsOur results demonstrate that miR-374-5p significantly downregulates both the protein and mRNA levels of its target gene PTEN, as confirmed by dual luciferase assays. qRT-PCR and Western blot analyses revealed that osteoblastic markers-including Runx2, OSX, and OCN-were markedly reduced in the miR-374-5p mimic group, whereas an opposite trend was observed in the inhibitor group. In vitro, overexpression of miR-374-5p suppressed osteoblast differentiation, as evidenced by decreased calcium nodule formation and reduced ALP activity compared to controls. Furthermore, co-transfection of miR-374-5p mimics with the PI3K/AKT pathway inhibitor LY294002 in osteoblasts led to significantly lower expression of PI3K/AKT pathway-related genes, and notably, the inhibitory effect of miR-374-5p on osteoblast differentiation was reversed by LY294002 treatment.
conclusionOur findings indicate that miR-374-5p inhibits osteogenesis in rat osteoblasts by targeting PTEN and modulating the PI3K/AKT signaling pathway.
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