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Circ_FBLN1 promotes the proliferation and osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by regulating let-7i-5p/FZD4 axis and Wnt/β-catenin pathway

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Journal of Bioenergetics and Biomembranes Aims and scope Submit manuscript

Abstract

Background

Recently, more and more circular RNAs (circRNAs) have been identified in osteogenesis. In this study, we aimed to explore the effect of circ_FBLN1 on the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs).

Methods

The protein levels of osteogenesis-related genes, let-7i-5p, frizzled class receptor 4 (FZD4), Ki67, Wnt6 and β-catenin were measured by western blot assay. The levels of circ_FBLN1, FBLN1 mRNA and FZD4 mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR) assay. The feature of circ_FBLN1 was investigated by RNase R and Actinomycin D assays. Cell proliferation ability was evaluated by colony formation assay and 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The targeting relationship between let-7i-5p and circ_FBLN1 or FZD4 was verified by dual-luciferase reporter assay.

Results

Circ_FBLN1 level was enhanced during the osteogenic differentiation of hBMSCs. Silencing of circ_FBLN1 repressed cell proliferation and osteogenic differentiation in hBMSCs. For mechanism analysis, circ_FBLN1 was found to act as a sponge for let-7i-5p and FZD4 served as a direct target gene of let-7i-5p. Let-7i-5p was downregulated during the osteogenic differentiation of hBMSCs and let-7i-5p inhibition restored the effects of circ_FBLN1 knockdown on the proliferation and osteogenesis of hBMSCs. Moreover, let-7i-5p overexpression suppressed cell proliferation and osteogenesis in hBMSCs through targeting FZD4. In addition, circ_FBLN1 knockdown reduced the levels of Wnt6 and β-catenin in hBMSCs, indicating the inactivation of Wnt/β-catenin pathway.

Conclusion

Knockdown of circ_FBLN1 inhibited the proliferation and osteogenesis of hBMSCs by regulating let-7i-5p/FZD4 axis and repressing Wnt/β-catenin pathway.

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Data availability

The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.

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Authors and Affiliations

  1. Department of Spine, Zaozhuang Municipal Hospital, Zaozhuang City, Shandong Province, China

    Zilong Zhang

  2. Department of the Orthopaedic Trauma, Zaozhuang Mining Group Zaozhuang Hospital, Zaozhuang, Shandong, China

    Huachao Zhou & Fei Sun

  3. Department of the Orthopaedic Trauma, Qingdao Central Hospital, Qingdao, Shandong, China

    Jianjian Han

  4. Department of Orthopedics, No.4 Hospital Beijing University of Chinese Medicine, No.202 Xuezhuang Community, Zhongxin Street, Zaozhuang, 277101, Shandong, China

    Yongyuan Han

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  1. Zilong Zhang
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  2. Huachao Zhou
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  3. Fei Sun
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  4. Jianjian Han
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Correspondence to Yongyuan Han.

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Zilong Zhang and Huachao Zhou contributed equally to this work

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Zhang, Z., Zhou, H., Sun, F. et al. Circ_FBLN1 promotes the proliferation and osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by regulating let-7i-5p/FZD4 axis and Wnt/β-catenin pathway. J Bioenerg Biomembr 53, 561–572 (2021). https://doi.org/10.1007/s10863-021-09917-0

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  • DOI: https://doi.org/10.1007/s10863-021-09917-0

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