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METTL3 m6A-dependently promotes miR-21-5p maturation to accelerate choriocarcinoma progression via the HIF1AN-induced inactivation of the HIF1A/VEGF pathway

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Abstract

Background

Gestational choriocarcinoma is a highly malignant neoplastic disease derived from pathological changes in trophoblastic cells. Recent evidences have shown that N6-methyladenosine (m6A) modifications play important role in modulating the development of multiple cancers, but the detailed mechanisms by which m6A-mediated choriocarcinoma progression have not been fully delineated.

Objectives

This study aimed to investigate the role of m6A in choriocarcinoma and reveal its underlying molecular mechanisms.

Methods

The expression of METTL3, miR-21-5p and HIF1AN was detected using RT-qPCR in tissues and cells. The protein expression of METTL3, HIF1AN, HIF1A and VEGF were measured by western blot. The luciferase reporter assays and RNA immunoprecipitation (RIP) were used to verify the relationship between miR-21-5p and HIF1AN. The CCK-8, colony formation and transwell assays were used to detected cell proliferation and cell migration, respectively.

Results

Here, we demonstrated that the m6A methyltransferase-like 3 (METTL3) was aberrantly high-expressed in the clinical choriocarcinoma tissues and choriocarcinoma cell lines compared to the corresponding normal counterparts. The following functional experiments verified that silencing of METTL3 suppressed cell proliferation, migration, epithelial-mesenchymal transition (EMT) and tumorigenesis in vitro and in vivo to hamper the aggressiveness of choriocarcinoma. Next, the mechanical experiments confirmed that METTL3 promoted the maturation of miR-21-5p in an m6A-dependent manner, and elevated miR-21-5p subsequently degraded its downstream hypoxia-inducible factor asparagine hydroxylase (HIF1AN) by targeting its 3ʹ untranslated regions (3ʹ-UTR), resulting in the activation of the tumor-promoting HIF1A/VEGF pathway. Finally, the rescuing experiments verified that METTL3 ablation-induced inhibitory effects on the malignant phenotypes in choriocarcinoma were all abrogated by both miR-21-5p overexpression and HIF1AN downregulation.

Conclusions

Collectively, this study firstly reported the involvement of the METTL3/m6A/miR-21-5p/HIF1AN signaling cascade in regulating the progression of choriocarcinoma, which provided novel biomarkers for the diagnosis and treatment of this disease.

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Data availability statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This study was supported by the Doctor Research Fund of The First People’s Hospital of Yunnan Province (KHBS-2020-003) and the Yunnan Provincial Department of Science and Technology -Kunming Medical University Joint Special Project on Applied Basic Research (202201AY070001-229).

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Author notes

  1. Kefan Ye and Lingchuan Li contributed equally to this article

Authors and Affiliations

  1. Department of Gynaecology, The First People’s Hospital of Yunnan Province, Kunming, 650032, China

    Kefan Ye, Lingchuan Li & Dongjie Wang

  2. The Affiliated Hospital of Kunming University of Science and Technology, No. 157, Jinbi Road, Xishan District, Kunming, 650032, Yunnan, China

    Kefan Ye, Lingchuan Li & Dongjie Wang

  3. Guanganmen Hospital Southern District, China Academy of Chinese Medical Science, Cardiovascular Division, Beijing, 102600, China

    Bao Wu

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  1. Kefan Ye
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Contributions

DW conceived of the study and KY and LL carried out its design. KY, LL and BW performed the experiments. BW collected the clinical samples. BW and LL analyzed the data and KY wrote the paper. DW revised the paper. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Dongjie Wang.

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Conflict of interest

Kefan Ye, Lingchuan Li, Bao Wu and Dongjie Wang declared that they have no relevant conflicts of interest.

Ethical approval

This study was approved by the Ethics Committee of The First People's Hospital of Yunnan Province (KHLL2021-121), and all participants signed an informed consent before tissues sample collection. All animal experiments were approved by the approval of Animal Ethics Committee of The First People's Hospital of Yunnan Province.

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Ye, K., Li, L., Wu, B. et al. METTL3 m6A-dependently promotes miR-21-5p maturation to accelerate choriocarcinoma progression via the HIF1AN-induced inactivation of the HIF1A/VEGF pathway. Genes Genom 44, 1311–1322 (2022). https://doi.org/10.1007/s13258-022-01309-x

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