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LncRNA LOXL1-AS1 controls osteogenic and adipocytic differentiation of bone marrow mesenchymal stem cells in postmenopausal osteoporosis through regulating the miR-196a-5p/Hmga2 axis

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Abstract

Introduction

Exploring molecular mechanisms of human bone marrow mesenchymal stem cells (hBMMSCs) differentiation, a crucial step for bone formation, is a new direction for treating postmenopausal osteoporosis. LncRNAs are involved in lots of biological processes including hBMMSCs differentiation. The present study aimed to explore the effect of LOXL1-AS1 on hBMMSCs differentiation.

Materials and methods

We examined the expression levels of LOXL1-AS1, miR-196a-5p and Hmga2 in peripheral blood from postmenopausal osteoporosis patients by RT-qPCR, and detected their changes during the osteogenic differentiation of hBMMSCs by RT-qPCR. RT-qPCR and western blot measured the level of biomarkers of bone formation and osteogenic differentiation (osteopontin, OPN; Alkaline phosphatase, ALP; Runt-related transcription factor-2, Runx-2). The effects of LOXL1-AS1 on the osteogenic and adipocytic differentiation of hBMMSCs were, respectively, determined by ALP, ARS staining assays and oil red O staining assay.

Results

The abnormal high expression of LOXL1-AS1 was found in patients. LOXL1-AS1 expression showed a gradual decrease during the osteogenic differentiation of hBMMSCs. However, LOXL1-AS1 overexpression inhibited the hBMMSCs osteogenic differentiation but promoted adipocytic differentiation. Furthermore, LOXL1-AS1 was identified to be a sponge of miR-196a-5p and Hmga2 as a target gene of miR-196a-5p. Besides, LOXL1-AS1 sponged miR-196a-5p to mediate Hmga2 expression, which played contrary effects on regulating osteogenic and adipocytic differentiation of hBMMSCs. Moreover, LOXL1-AS1/miR-196a-5p/Hmga2 axis regulated hBMMSCs differentiation through controlling C/EBPβ-mediated PPARγ expression.

Conclusion

These findings facilitate understanding the molecular mechanism of hBMMSCs differentiation and bring up a novel sight for postmenopausal osteoporosis therapy.

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Data availability

All data generated or analyzed during this study are included in this published article.

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Funding

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Author information

Authors and Affiliations

  1. Department of Geriatrics, the First Affiliated Hospital, Sun Yat-Sen University, No. 58 Zhongshan 2nd Road, Guangzhou City, 510080, Guangdong Province, China

    Ling Zhang

  2. Department of Ultrasound, Peking University Shenzhen Hospital, Shenzhen City, 518036, Guangdong Province, China

    Haiqin Xie

  3. Healthcare Office, the First Affiliated Hospital, Sun Yat-Sen University, No. 58 Zhongshan 2nd Road, Guangzhou City, 510080, Guangdong Province, China

    Shiliang Li

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  1. Ling Zhang
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Contributions

LZ conceived and designed the experiments, HQX analyzed and interpreted the results of the experiments, SLL performed the experiments.

Corresponding author

Correspondence to Ling Zhang.

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Conflict of interest

The authors state that there are no conflicts of interest to disclose.

Ethical approval

We obtained written informed consent from all participants. Our study obtained the approval from the Ethics Committee of the First Affiliated Hospital, Sun Yat-Sen University.

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Ling Zhang and Haiqin Xie contributed equally to the work.

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774_2020_1123_MOESM1_ESM.tif

Supplementary file1 Supplementary figure 1. Expression of miRNAs interacting with LOXL1-AS1. The expression levels of miR-28-5p, miR-423-5p and miR-143-3p were determined using RT-qPCR in peripheral blood from postmenopausal osteoporosis patients and healthy postmenopausal females (control). *p < 0.01 versus control. (TIF 736 kb)

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Zhang, L., Xie, H. & Li, S. LncRNA LOXL1-AS1 controls osteogenic and adipocytic differentiation of bone marrow mesenchymal stem cells in postmenopausal osteoporosis through regulating the miR-196a-5p/Hmga2 axis. J Bone Miner Metab 38, 794–805 (2020). https://doi.org/10.1007/s00774-020-01123-z

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  • DOI: https://doi.org/10.1007/s00774-020-01123-z

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