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USF2 reduces BMP3 expression via transcriptional activation of miR-34a, thus promoting osteogenic differentiation of
Han-Bing Zeng1, Li-Qiang Dong1, Yu-Liang Huang1
1Department of Orthopaedics, The Second Affiliated Hospital of Zhejiang Chinese Medical University, No.318, Chaowang Road, Gongshu District, Hangzhou, 310005, Zhejiang Province, China.
Introduction:
Osteoporosis is the most susceptible disease for people over 60. The main cause of osteoporosis is the decreased osteogenic differentiation of mesenchymal stem cells (MSCs). Here we showed that upstream stimulatory factor 2 (USF2)/microRNA-34a (miR-34a)/bone morphogenetic protein 3 (BMP3) axis regulated osteogenic differentiation of BMSCs.
Materials And Methods:
USF2 and miR-34a expression were examined using qPCR. Protein levels of BMP3 and osteogenic markers expression were evaluated using both western blot and qPCR. Activity of ALP was determined by ALP assay kit. Mineralization capacity of hBMSCs was assessed using ARS. Besides, CHIP assay was employed to verify whether USF2 could bind to miR-34a promoter. Finally, RIP assay and dual-luciferase reporter assay were employed to verify whether miR-34a directly bound to BMP3.
Results:
Our results suggested that miR-34a was upregulated during osteogenic differentiation of BMSCs, and miR-34a overexpression could enhance osteogenic differentiation of BMSCs. USF2 could positively regulate miR-34a expression by interacting with its promoter. USF2 overexpression enhanced osteogenic differentiation of BMSCs, while miR-34a inhibition reversed the effect. Besides, BMP3 was the target of miR-34a. MiR-34a overexpression enhanced osteogenic differentiation of BMSCs, which was abolished by BMP3 overexpression.
Conclusion:
Taken together, USF2 enhanced osteogenic differentiation of BMSCs via downregulating BMP3 by interacting with miR-34a promoter.
Insights
Upstream stimulatory factor 2 (USF2) enhances bone formation in mesenchymal stem cells (MSCs) by regulating microRNA-34a (miR-34a) and downregulating bone morphogenetic protein 3 (BMP3). This discovery offers new insights into treating osteoporosis.
Area of Science:
- Biomedical research
- Stem cell biology
- Molecular biology
Background:
- Osteoporosis is a significant health concern for individuals over 60.
- Reduced osteogenic differentiation of mesenchymal stem cells (MSCs) is a primary cause of osteoporosis.
- The USF2/miR-34a/BMP3 axis's role in regulating BMSC osteogenic differentiation is investigated.
Purpose of the Study:
- To elucidate the regulatory mechanism of osteogenic differentiation in BMSCs.
- To investigate the role of the USF2/miR-34a/BMP3 signaling pathway.
Main Methods:
- Quantitative PCR (qPCR) for gene expression analysis.
- Western blot for protein level assessment.
- ALP activity assay, Alizarin Red S staining for mineralization.
- Chromatin immunoprecipitation (CHIP) and RNA immunoprecipitation (RIP) assays.
Main Results:
- miR-34a expression increased during BMSC osteogenic differentiation, enhancing the process.
- USF2 positively regulated miR-34a, promoting osteogenic differentiation.
- BMP3 was identified as a direct target of miR-34a.
- USF2 promoted osteogenic differentiation by downregulating BMP3 via miR-34a.
Conclusions:
- USF2 enhances BMSC osteogenic differentiation by downregulating BMP3 through miR-34a.
- This pathway represents a potential therapeutic target for osteoporosis.
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