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MiR-135-5p Alleviates Bone Cancer Pain by Regulating Astrocyte-Mediated Neuroinflammation in Spinal Cord through JAK2/STAT3 Signaling Pathway

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Abstract

Bone cancer pain (BCP) was associated with microRNA dysregulation. In this study, we intended to clarify the potential role of miR-135-5p in a BCP mouse model, which was established by tumor cell implantation (TCI) in the medullary cavity of the mouse femur. The BCP-related behaviors were tested, including the paw withdrawal mechanical threshold (PWMT) and number of spontaneous flinches (NSF). The miRNA expression profiles in astrocytes of the sham and tumor groups were compared, and miRNA microarray and quantitative real-time PCR (qRT-PCR) assays confirmed that the amount of expression of miR-135-5p was significantly decreased in astrocytes of the tumor group. Gain- and loss-of-function studies showed that miR-135-5p could inhibit astrocyte activation and inflammation cytokine (TNF-α and IL-1β) expression. The relation between miR-135-5p and JAK2 was detected by bioinformatic analysis and dual luciferase reporter gene assay. By conducting in vitro experiments, it was shown that the miR-135-5P mimics lowered the level of JAK2/STAT3 proteins and inflammatory factors in astrocytes. Moreover, in vivo analysis on BCP mice model indicated that the miR-135-5p agonist could sufficiently increase PWMT and decrease NSF. Meanwhile, reduced activation of astrocytes in the spinal cord, as well as decreased expression of JAK2/STAT3 and inflammatory mediators, were found after miR-135-5p agonist treatment. Collectively, the results showed that miR-135-5p could potentially reduce BCP in mice through inhibiting astrocyte-mediated neuroinflammation and blocking of the JAK2/STAT3 signaling pathway, indicating that the upregulation of miR-135-5P could be a therapeutic focus in BCP treatment.

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Data Availability

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

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Acknowledgements

The authors would like to thank those previous researchers who updated their data onto the databases employed in the present study.

Funding

This work is supported by the Fundamental Research Funds for the Central Universities (No. 2042019kf0073).

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Author notes

  1. Ming Liu and Xuefeng Cheng contributed equally to this work.

Authors and Affiliations

  1. Department of Anaesthesiology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Ming Liu, Xuefeng Cheng, Hong Yan, Jingli Chen & Caihua Liu

  2. Department of Spine Surgery, Renmin Hospital of Wuhan University, No. 9 ZhangZhiDong Street, Wuchang District, Wuhan, Hubei, China

    Zhonghui Chen

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  1. Ming Liu
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  2. Xuefeng Cheng
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  3. Hong Yan
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Contributions

M.L., X.F.C., and Z.H.C. designed the experiments. M.L. and X.F.C. collected samples. M.L., X.F.C., and H.Y. conducted the experiments and acquired the data. M.L., H.Y., and J.L.C. analyzed the data. M.L. and X.F.C. wrote the manuscript. H.Y., J.L.C., and C.H.L. revised the manuscript. All authors approved the final version. The corresponding author had full access to all the data in the study and had final responsibility for the decision to submit for publication.

Corresponding author

Correspondence to Zhonghui Chen.

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All experimental approaches were reviewed and approved by the Institutional Animal Care and Use Committee of the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology.

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Liu, M., Cheng, X., Yan, H. et al. MiR-135-5p Alleviates Bone Cancer Pain by Regulating Astrocyte-Mediated Neuroinflammation in Spinal Cord through JAK2/STAT3 Signaling Pathway . Mol Neurobiol 58, 4802–4815 (2021). https://doi.org/10.1007/s12035-021-02458-y

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