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Hirschsprung’s disease: m6A methylase VIRMA suppresses cell migration and proliferation by regulating GSK3β

Pediatric Research (2024)Cite this article

Abstract

Background

N6-methyladenosine (m6A) is the most abundant mRNA modification in mammals, participating in various biological processes. VIRMA is a key methyltransferase involved in m6A modification. However, the role of VIRMA in Hirschsprung’s disease (HSCR) remains unclear. This study aims to investigate the function of VIRMA in HSCR and identify its corresponding regulatory mechanisms.

Methods

The expression of VIRMA and GSK3β in colon tissues of HSCR was examined using RT-qPCR, Western blot, and Immunohistochemistry. Immunofluorescence detected localization of VIRMA and GSK3β. Cell proliferation was measured by CCK8 and EdU assays, and cell migration was evaluated via cell migration and wound healing assays. The stability of GSK3β mRNA was assessed using the actinomycin D assay and the overall level of m6A in cells was assessed by colorimetric assay.

Results

VIRMA was significantly downregulated in narrow-segment colon tissue. Silencing of VIRMA inhibited cell proliferation and migration. VIRMA can inhibit the degradation of GSK3β mRNA and increase the expression of GSK3β. GSK3β was significantly upregulated in narrow-segment colon tissues. Accordingly, our findings showed that GSK3β mediated the VIRMA-driven cell migration and proliferation.

Conclusion

VIRMA can inhibit cell migration and proliferation by upregulating the expression of GSK3β, contributing to the onset of HSCR.

Impact

  • The expressions of VIRMA were significantly reduced in HSCR, while GSK3β expression was increased in HSCR, and can be used as a molecular marker.

  • VIRMA overexpression promoted the proliferation and migration of SH-SY5Y and HEK-293T cells.

  • VIRMA can inhibit the degradation of GSK3β mRNA and increase the expression of GSK3β.

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Fig. 1: VIRMA expression was reduced in the HSCR diseased segment colon tissues.
Fig. 2: VIRMA inhibited the proliferation of human 293T and SH-SY5Y cells.
Fig. 3: VIRMA inhibited the migration of human 293 T and SH-SY5Y cells.
Fig. 4: VIRMA regulated the stability of GSK3β mRNA.
Fig. 5: Tissue samples substantiated a negative correlation between GSK3β and VIRMA.
Fig. 6: VIRMA inhibited the proliferation and migration of human 293T and SH-SY5Y cells by targeting GSK3β.

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Data availability

Data included in this manuscript are available upon request by contacting the corresponding author and will be freely available to any researcher wishing to use them for non-commercial purposes without breaching participant confidentiality.

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Funding

This study was supported by the Guangdong Basic and Applied Basic Research Foundation of China (2019A1515011086).

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Author notes

  1. These authors contributed equally: Yang Yang, Mengzhen Zhang, Nan Li.

Authors and Affiliations

  1. Department of Pediatric Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou, 510282, Guangdong, China

    Yang Yang, Mengzhen Zhang, Nan Li, Chen Wang, Huirong Yang, Xinwei Hou, Jiaming Yang, Kaisi Fan, Liucheng Yang & Kai Wu

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Contributions

Yang Yang, Mengzhen Zhang, Nan Li: preparation of the manuscript, statistics, and writing the manuscript. Chen Wang, Huirong Yang, Xinwei Hou: literature search, literature analyses. Jiaming Yang, Kaisi Fan: literature analyses and reviewed the manuscript. Kai Wu, Liucheng Yang: guide the project and revise the manuscript. All authors contributed to the article and approved the submitted version.

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Correspondence to Kai Wu.

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The studies involving human participants were reviewed and approved by the Ethics Committee of Zhujiang Hospital of Southern Medical University. Written informed consent was obtained from patients’ parents for the publication of any potentially identifiable images or data included in this article.

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Yang, Y., Zhang, M., Li, N. et al. Hirschsprung’s disease: m6A methylase VIRMA suppresses cell migration and proliferation by regulating GSK3β. Pediatr Res (2024). https://doi.org/10.1038/s41390-024-03136-0

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  • DOI: https://doi.org/10.1038/s41390-024-03136-0

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