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Exendin-4 Reversed the PC12 Cell Damage Induced by circRNA CDR1as/miR-671/GSK3β Signaling Pathway

Journal of Molecular Neuroscience volume 71, pages 778–789 (2021)Cite this article

Abstract

The purpose of this paper is to study the effect of circRNA cerebellar degeneration–related protein 1 antisense RNA(CDR1as)/miR-671/GSK3β signaling pathway on PC12 cell injury and the mechanism of Exendin-4 (Ex-4) in PC12 cell injury protection. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to detect the expression levels of circular RNA CDR1as and miR-671 in PC12 cells. By overexpressing or knocking out CDR1as, miR-671, and GSK3β, the role of CDR1as, miR-671, and GSK3β in PC12 cell injury was analyzed. The binding of CDR1as to miR-671 and GSK3β to miR-671 was verified by dual luciferase reporter assay. PC12 cells were treated with 1-methyl-4 phenyl-pyridine ion (MPP+) to construct a PC12 cell damage model. PC12 cell transfection experiments were used to confirm the role of CDR1as/miR-671/GSK3β signal axis in PC12 cell damage, and the role of Ex-4 in the association of circRNA CDR1as/miR-671/GSK3β signaling axis and PC12 cell damage. PC12 cell damage was detected by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and cellular lactate dehydrogenase (LDH) release. Ex-4 reversed the phosphorylation levels of PI3K, AKT, and GSK-3β in MPP+-treated PC12 cells, and reduced MPP+-induced PC12 cell damage. CircRNA CDR1as upregulated the expression of GSK3β by sponge miR-671. Ex-4 downregulated CDR1as expression and upregulated miR-671 expression in MPP+-induced PC12 cell. Silencing of CDR1as reduced MPP+-induced PC12 cell damage. CDR1as transfection downregulated the expression of miR-671 in PC12 cells, promoted the expression and phosphorylated of GSK3β, and induced PC12 cell damage. GSK3β silencing reversed CDR1as-induced PC12 cell damage. CDR1as promoted the phosphorylation level of GSK3β in PC12 cells to cause cell damage; Ex-4 reversed the phosphorylation of GSK3β caused by CDR1as in PC12 cells and reduced the PC12 cell damage caused by CDR1as. Ex-4 reverses the damage of PC12 cells induced by CDR1as/miR-671/GSK3β signaling pathway.

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Funding

This work was supported by the Shanghai Health and Family Planning Commission Clinical Research Project for Health Industry (No.: 20184Y0199).

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Author notes

  1. Hui Quan and Qiudan Chen contributed equally to this work.

Affiliations

  1. Department of Neurology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, 200072, China

    Hui Quan

  2. Department of Central Laboratory, Clinical Laboratory, Jing’an District Center Hospital of Shanghai, Fudan University, Shanghai, 200040, China

    Qiudan Chen

  3. Department of Central Laboratory, Clinical Medicine Scientific and Technical Innovation Park, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, 200435, China

    Kesheng Wang, Qinwan Wang, Meiling Lu, Yue Zhang, Sujing Qiang, Shaobo Xue, Lin Han, Huanhuan Zhu, Qiangyuan Fan, Meiyu Bao & Weifeng Wang

  4. Department of Laboratory Medicine, Jiading District Central Hospital Affiliated Shanghai University of Medicine & Health Sciences, Shanghai, 201800, China

    Weixin Xu

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  1. Hui Quan
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  2. Qiudan Chen
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Correspondence to Weixin Xu or Weifeng Wang.

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Quan, H., Chen, Q., Wang, K. et al. Exendin-4 Reversed the PC12 Cell Damage Induced by circRNA CDR1as/miR-671/GSK3β Signaling Pathway. J Mol Neurosci 71, 778–789 (2021). https://doi.org/10.1007/s12031-020-01698-2

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  • DOI: https://doi.org/10.1007/s12031-020-01698-2

Keywords

  • Exendin-4
  • Circular RNA
  • circRNA cerebellar degeneration–related protein 1 antisense RNA
  • GSK3β
  • miR-671