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miR-346-3p promotes osteoclastogenesis via inhibiting TRAF3 gene

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Abstract

MicroRNAs (miRNAs) modulate gene expression and regulate many physiological and pathological conditions. However, their modulation and effect in osteoclastogenesis remain unknown. In this study, we investigated the role of miR-346-3p in regulating the osteoclast differentiation from RAW264.7 cells. We used the miRNA microarray assay, miR-346-3p mimic transfection, tartrate resistant acid phosphatase (TRAP) staining, bone resorption assay, qRT-PCR, and western blot. Our results showed that the expression of miR-346-3p was significantly upregulated during osteoclast differentiation. Further, by transfecting cells with miR-346-3p mimic, we observed an increased number of TRAP-positive multinucleated cells, increased pit area caused by bone resorption, and enhanced expression of osteoclast-specific genes and proteins. Conversely, miR-346-3p inhibition attenuated the osteoclast differentiation and function. Software-mediated prediction and validation using luciferase reporter assay showed that TRAF3, a negative regulator of osteoclast differentiation, was inhibited by miR-346-3p overexpression. Our results showed that miR-346-3p directly targeted TRAF3 mRNA via binding to its 3′-UTR and inhibited the expression of TRAF3 protein. Taken together, our results revealed that miR-346-3p promotes the regulation of osteoclastogenesis by suppressing the TRAF3 gene. In conclusion, miR-346-3p could be a novel therapeutic target for bone loss-related pathogenesis.

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Acknowledgements

We would like to appreciate lab members for their support to our research and thank Editage (www.editage.cn) for English language editing.

Funding

This study was supported by grants from the National Natural Science Foundation of China (31700854), the Project of Natural Science Foundation of Anhui Province (1908085MH276), the Key Program of Anhui Educational Committee (KJ2018A1011 and KJ2019A0392), the Scientific Research Foundation of Bengbu Medical College (BYKY17118, BYKY18108, and BYKY1848ZD), the Translational Medicine Key Projects of Bengbu Medical College (BYTM2019006 and BYTM2019012), the Scientific Research Innovation Team of Bengbu Medical College (BYKC201910), the Postgraduate Research Innovation Project of Bengbu Medical College (Byycx1827, Byycx1954, Byycx1956, and Byycz1929), and the National Training Programs of Innovation and Entrepreneurship for Undergraduate (201810367013 and 201810367028).

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Author notes

  1. Yingji Mao and Yu Chen contributed equally to this work.

Authors and Affiliations

  1. School of Life Science, Bengbu Medical College, Bengbu, 233030, People’s Republic of China

    Yingji Mao, Yu Chen, Yingxiao Fu, Mengxiang Liang, Yansong Zhu, Fugen Yang, Feixiang Li, Zhe Zhang & Chuanfeng Wan

  2. Anhui Province Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu, 233030, People’s Republic of China

    Yingji Mao & Pinghui Zhou

  3. Department of Plastic Surgery, First Affiliated Hospital of Bengbu Medical College, Bengbu, 233000, People’s Republic of China

    Yu Chen

  4. Department of Orthopedics, First Affiliated Hospital of Bengbu Medical College, Bengbu, 233000, People’s Republic of China

    Jingjing Guan & Pinghui Zhou

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  1. Yingji Mao
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  2. Yu Chen
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Correspondence to Pinghui Zhou.

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The authors declare that they have no competing interests.

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Editor: Tetsuji Okamoto

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Mao, Y., Chen, Y., Fu, Y. et al. miR-346-3p promotes osteoclastogenesis via inhibiting TRAF3 gene. In Vitro Cell.Dev.Biol.-Animal 56, 533–542 (2020). https://doi.org/10.1007/s11626-020-00479-w

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  • DOI: https://doi.org/10.1007/s11626-020-00479-w

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