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Long non-coding RNA GAS5, by up-regulating PRC2 and targeting the promoter methylation of miR-424, suppresses multiple malignant phenotypes of glioma

Journal of Neuro-Oncology volume 148, pages 529–543 (2020)Cite this article

Abstract

Purpose

Malignant gliomas remain significant challenges in clinic and pose dismal prognosis on patients. In this study, we focused on growth arrest-specific 5 (GAS5), a tumor suppressive long non-coding RNA in glioma, explored its crosstalk with miR-424, and examined their biological functions in glioma.

Methods

Expressions of GAS5 and miR-424 were measured using qRT-PCR. The regulation of GAS5 on miR-424 expression was examined in GAS5-overexpressing glioma cells by combining methylation-specific PCR, western blotting, and RNA immunoprecipitation. Functional significance of GAS5 and miR-424 on in vitro cell proliferation, apoptosis, migration, invasion, and in vivo tumor growth was examined using colony formation, flow cytometry, wound healing, transwell assay, and the xenograft model, respectively. The potential targeting of AKT3 by miR-424 was investigated using luciferase reporter assay.

Results

GAS5 and miR-424 were significantly down-regulated in glioma cells. GAS5 directly interacted with enhancer of zeste homolog 2 (EZH2), stimulated the formation of polycomb repressive complex 2 (PRC2), reduced the levels of DNA methyltransferases (Dnmts), alleviated promoter methylation of miR-424, and promoted miR-424 expression. Functionally, GAS5, by up-regulating miR-424, inhibited cell proliferation, migration, and invasion, while increased apoptosis of glioma cells in vitro, and suppressed xenograft growth in vivo. miR-424 directly inhibited AKT3 and altered the expressions of AKT3 targets, cyclinD1, c-Myc, Bax, and Bcl-2, which might contribute to its tumor suppressive activities.

Conclusions

GAS5, by inhibiting methylation and boosting expression of miR-424, inhibits AKT3 signaling and suppresses multiple malignant phenotypes. Therefore, stimulating GAS5/miR-424 signaling may benefit the treatment of glioma.

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Data availability

All data generated or analyzed during this study are included in this published article.

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Affiliations

  1. Department of Neurosurgery, Xiangya Hospital, Central South University, No.87, Xiangya Road, Changsha, 410008, P.R. China

    Chen Jin, Jie Zhao, Zhi-Ping Zhang, Ming Wu, Jian Li, Ge-Lei Xiao, Bo Liu, Yu-Xiang Liao & Jing-Ping Liu

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  1. Chen Jin
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  2. Jie Zhao
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  3. Zhi-Ping Zhang
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Contributions

CJ and JPL contrinutrd in guarantor of integrity of the entire study, study concepts, study design, definition of intellectual content, literature research. JZ, ZPZ, MW, JL, GLX, YXL and BL contributrd in clinical studies. CJ contributrd in experimental studies, data analysis, manuscript editing and manuscript review. JPL, CJ, JZ, and ZPZ contributed in data acquisition. CJ and GLX contributed in statistical analysis. CJ, JPL, JZ and JL contributed in manuscript preparation.

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Correspondence to Jing-Ping Liu.

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Jin, C., Zhao, J., Zhang, ZP. et al. Long non-coding RNA GAS5, by up-regulating PRC2 and targeting the promoter methylation of miR-424, suppresses multiple malignant phenotypes of glioma. J Neurooncol 148, 529–543 (2020). https://doi.org/10.1007/s11060-020-03544-2

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Keywords

  • Glioma
  • lncRNA-GAS5
  • PRC2
  • miR-424
  • Methylation