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AKT/GSK-3β/β-catenin signaling pathway participates in erythropoietin-promoted glioma proliferation

  • Laboratory Investigation
  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

Purpose

Although erythropoietin (EPO) has been proven to significantly promote the proliferation of cancer cells, the mechanism for promoting glioma proliferation is poorly understood. Here, we examined the functional role of the AKT/GSK-3β/β-catenin signaling pathway in the EPO-mediated proliferation of glioma.

Methods

The distribution of EPO and Ki-67 among clinical samples with different WHO grades was plotted by Immunological Histological Chemistry analysis. U87 and U251 glioma cell lines were treated with short hairpin RNA targeting (shEPO), recombinant human erythropoietin (rhEPO) and/or AKT-specific inhibitor (MK-2206). The changes in phosphorylated AKT, nuclear β-catenin, cyclin D1 and p27kip1 expression were detected. Cell cycle distributions and glioma proliferation in vitro and in vivo were analyzed.

Results

The expression level of EPO was significantly elevated with the increase of WHO grade and Ki67 in clinical glioma specimens. In vitro, knockdown of endogenous EPO in U87 and U251 cells effectively block the phosphorylation of AKT and GSK-3β and the expression of nuclear β-catenin. shEPO treatment also significantly decreased the expression of cyclin D1 and increased the expression of p27kip1. The cell cycle transition then slowed down and the proliferation of glioma cells or mouse xenograft tumors both decreased. Treatment of cells or tumors with extra rhEPO reversed the above biological effects mediated by shEPO. rhEPO-induced activation of the AKT/GSK-3β/β-catenin pathway and proliferation were abolished by MK-2206.

Conclusions

Our study identified the AKT/GSK-3β/β-catenin axis as a critical mediator of EPO-induced glioma proliferation and further provided a clinically significant dimension to the biology of EPO.

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Data availability

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

Abbreviations

EPO:

Erythropoietin

GSK-3β:

Glycogen synthase kinase-3β

rhEPO:

Recombinant human erythropoietin

siRNA:

Short interfering RNA

MK:

MK-2206

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Acknowledgements

We would like to thank Professor Xinggang Zhi for his technical assistance.

Funding

This research was supported by the National Natural Science Funds of China under Awards Nos. 81301630, 81401070 and 81771355, the Science and Technology Research Project of Chongqing Municipal Education Commission, China (Award No. KJ1500219), and the Science and Technology Project of Yuzhong District, Chongqing, China (Award No. 20150108).

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Authors and Affiliations

  1. Department of Neurosurgery, The First Affiliated Hospital of Chongqing Medical University, No. 1 Youyi Road, Chongqing, 400016, China

    Zhaohua Tang, Gang Yang, Xiaoshu Wang, Zhengbu Liao, Zili Liu, Min Fang, Wentao Wang, Xiaochuan Sun & Gang Huo

  2. Laboratory Animal Centre, Chongqing Medical University, No. 1 Youyi Road, Chongqing, 400016, China

    Feilan Chen

  3. Laboratory of Medical Molecular Biology, Chongqing Medical University, Chongqing, 400016, China

    Zhiwei Zhang & Wenxin Zeng

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  1. Zhaohua Tang
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Contributions

ZT and GH contributed to the conception and design. All authors had full access to the data and participated in the analysis and interpretation of the data. ZT was responsible for drafting the manuscript. All of the authors reviewed the manuscript before submission. All authors read and approved the final manuscript.

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Correspondence to Gang Huo.

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The authors declare that they have no competing interests.

Ethical approval

The primary human brain tumor specimens were obtained from patients who provided informed consent under protocols approved by the Institutional Review Board of the First Affiliated Hospital of Chongqing Medical University. Animal experiments were approved by the Medical Ethics Committee of the First Affiliated Hospital of Chongqing Medical University. All procedures involving mice conformed to the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health.

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Tang, Z., Yang, G., Wang, X. et al. AKT/GSK-3β/β-catenin signaling pathway participates in erythropoietin-promoted glioma proliferation. J Neurooncol 149, 231–242 (2020). https://doi.org/10.1007/s11060-020-03602-9

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