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CircRNA-23525 regulates osteogenic differentiation of adipose-derived mesenchymal stem cells via miR-30a-3p

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Abstract

Adipose-derived mesenchymal stem cells (ADSCs) are considered to be seed cells in bone tissue engineering and emerging evidence indicates that circular RNAs (circRNAs) function in the osteogenic differentiation of ADSCs. The mechanisms of osteoblastic differentiation of ADSCs from the perspective of circRNA modulation are examined in this study. First, circRNA-23525 was upregulated during osteoblastic differentiation of ADSCs. Second, overexpression of circRNA-23525 increased Runx2, ALP and OCN at both mRNA and protein levels. Alkaline phosphatase (ALP) and Alizarin Red staining indicated a similar tendency. Silencing circRNA-23525 produced the opposite effect. Bioinformatics analysis with luciferase assays confirmed that circRNA-23525 functioned as a sponge for miR-30a-3p. In the osteoblastic differentiation of ADSCs, the dynamic expression of miR-30a-3p and circRNA-23525 resulted in an opposite trend at 3, 7 and 14 days. Overexpression of circRNA-23525 downregulated miR-30a-3p and knockdown of circRNA-23525 promoted the expression of miR-30a-3p. Bioinformatics methods and luciferase assays suggested that miR-30a-3p modulated Runx2 expression by targeting 3′UTR. Knockdown of miR-30a-3p facilitated osteogenesis in ADSCs and enhancing miR-30a-3p interfered with the osteogenic process. Finally, circRNA-23525 overexpression significantly increased Runx2 expression, while co-transfection of miR-30a-3p mimics reversed it. Runx2 expression was decreased in circRNA-23525-knockdown ADSCs but expression was rescued by including the miR-30a-3p inhibitor in the osteoblastic process. ALP activity and mineralized bone matrix confirmed the function of circRNA-23525/miR-30a-3p in osteogenesis. Taken together, the current study demonstrated that circRNA-23525 regulates Runx2 expression via targeting miR-30a-3p and is thus a positive regulator in the osteoblastic differentiation of ADSCs.

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Funding

This study was supported by grants from the National Nature Science Foundation of China (Nos. 31570950, 10502037, and 31070833), the Science and Technology Foundation of Sichuan Province (Nos. 2017SZ0032, 2010GZ0225, 2011GZ0335, and 2009SZ0139) and National Key Research and Development Program of China (No. 2016YFC1101404).

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Author notes

  1. Zeyou Guo and Luyang Zhao equally contributed to this work.

Authors and Affiliations

  1. The State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, Sichuan, 610041, People’s Republic of China

    Zeyou Guo, Luyang Zhao, Suhui Ji, Ting Long, Yanling Huang, Rui Ju & Jie Long

  2. Department of Oral and Maxillofacial Surgery, West China College of Stomatology, Sichuan University, Chengdu, Sichuan, 610041, People’s Republic of China

    Zeyou Guo, Luyang Zhao, Suhui Ji, Yanling Huang, Rui Ju, Wei Tang, Weidong Tian & Jie Long

  3. Engineering Research Center of Oral Translational Medicine, Ministry of Education, Chengdu, 610041, People’s Republic of China

    Weidong Tian

  4. National Engineering Laboratory for Oral Regenerative Medicine, Chengdu, 610041, People’s Republic of China

    Weidong Tian

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  1. Zeyou Guo
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Correspondence to Jie Long.

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All applicable international, national and institutional guidelines for the care and use of animals were followed. All procedures in our studies involving animals were executed in accordance with a protocol approved by the Animal Research Committee of Sichuan University (Chengdu, China).

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Guo, Z., Zhao, L., Ji, S. et al. CircRNA-23525 regulates osteogenic differentiation of adipose-derived mesenchymal stem cells via miR-30a-3p. Cell Tissue Res 383, 795–807 (2021). https://doi.org/10.1007/s00441-020-03305-7

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  • DOI: https://doi.org/10.1007/s00441-020-03305-7

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