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Nfib promotes chondrocyte proliferation and inhibits differentiation by mildly regulating Sox9 and its downstream genes

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Abstract

Background

Chondrocyte proliferation and differentiation play pivotal roles in regulating cartilage formation, endochondral bone formation, and repair. Cartilage damage and underdevelopment may cause severe joint diseases. Various transcription factors regulate cartilage development. Nuclear factor 1 B (Nfib) is a transcription factor that plays a regulatory role in various organs. However, the effect and mechanism of Nfib on the proliferation and differentiation of chondrocytes in cartilage are still largely unknown.

Methods and results

In the present study, we investigated the gene expression patterns in primary chondrocytes with Nfib overexpression or silencing by RNA sequencing (RNA-seq) technology. The results showed that Nfib overexpression significantly up-regulated genes that are related to chondrocyte proliferation and extracellular matrix (ECM) synthesis and significantly down-regulated genes related to chondrocyte differentiation and ECM degradation. However, with Nfib silencing, the genes involved in promoting chondrocyte differentiation were significantly up-regulated, whereas those involved in promoting chondrocyte proliferation were significantly down-regulated. Furthermore, quantitative real-time PCR (qRT-PCR), western blot, alcian blue staining and immunofluorescence staining assays further confirmed that Nfib potentially promotes chondrocyte proliferation and extracellular synthesis but inhibits differentiation.

Conclusions

The molecular mechanism of Nfib in promoting chondrocyte proliferation and inhibiting differentiation was probably achieved by stimulating Sox9 and its downstream genes. Thus, this study adds new insights regarding the underlying molecular mechanism of transcriptional regulation in cartilage.

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Data availability

The RNA-seq data set are available in the NCBI Sequence Read Archive (SRA) database (Accession Number: PRJNA707136).

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (grant number 2018YFC1706605) and the National Natural Science Foundation of China (grant number 81702136).

Funding

This work was supported by the National Key Research and Development Program of China (Grant Number 2018YFC1706605), Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine (Grant Number ZYYCXTD-D-202001ZYYCXTD-D-202001) and the National Natural Science Foundation of China (Grant Number 81702136).

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Authors and Affiliations

  1. Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, China

    Daian Pan, Daqing Zhao & Baojin Yao

  2. Department of Thoracic Surgery, The First Hospital of Jilin University, Changchun, China

    Benxin Qian

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  1. Daian Pan
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by DP and BQ. The first draft of the manuscript was written by DP. The review and editing of the manuscript was written by BY. The resources and supervision were performed by DZ. All authors read and approved the final manuscript.

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Correspondence to Daqing Zhao or Baojin Yao.

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All procedures were performed in accordance with the guidelines of the Institutional Animal Ethics Committee of Changchun University of Chinese Medicine (No. ccucm-2017–0015).

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Pan, D., Qian, B., Zhao, D. et al. Nfib promotes chondrocyte proliferation and inhibits differentiation by mildly regulating Sox9 and its downstream genes. Mol Biol Rep 48, 7487–7497 (2021). https://doi.org/10.1007/s11033-021-06767-2

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