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Mechanism of METTL14-mediated m6A modification in non-small cell lung cancer cell resistance to cisplatin

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Abstract

Methyltransferase-like 14 (METTL14) mediates N6-methyladenosine (m6A) modification to influence cancer progression. This study aims to determine the mechanism of METTL14-mediated m6A in non-small cell lung cancer (NSCLC) cell resistance to cisplatin (DDP). METTL14, miR-19a-5p, RBM24, and AXIN1 expressions in NSCLC tissues/cells were determined. DDP-resistant cell line was obtained, followed by the interference of METTL14 expression. NSCLC cells were treated with DDP to establish a drug-resistant cell line, and METTL14 expression in cells was intervened. The IC50 of NSCLC cells to DDP was measured by CCK-8 assay. NSCLC cell proliferation and apoptosis were observed by clone formation assay and flow cytometry. The content of m6A in total RNA in tissues and cells of NSCLC patients was detected using m6A Methylation Quantification Kit. The expressions of DGCR8-bound pri-miR-19a and m6A-modified pri-miR-19a were detected. The binding relationships between miR-19a-5p and RBM24 and RBM24 and AXIN1 were validated using dual-luciferase assay and RNA immunoprecipitation. Finally, mouse xenograft tumor model was established to verify the role of METTL14 in vivo. METTL14 was highly expressed in NSCLC. METTL14 silencing diminished IC50 to DDP, repressed NSCLC cell proliferation, and enhanced apoptosis. METTL14-mediated m6A induced recognition and processing of pri-miR-19a by DGCR8, thus promoting the transition of pri-miR-19a to miR-19a-5p, repressing RBM24 expression, reducing the binding of RBM24 and AXIN1, and suppressing AXIN1 transcription. miR-19a-5p overexpression or RBM24/AXIN1 silencing abolished the effect of METTL14 silencing on NSCLC cell resistance to DDP. METTL14 silencing in vivo enhanced the suppressive role of DDP to tumor growth. Collectively, METTL14-mediated m6A modification facilitated NSCLC cell resistance to DDP via miR-19a-5p/RBM24/AXIN1 axis.

Key messages

• METTL14 is highly expressed NSCLC and further increased in DDP-resistant cells.

• METTL14 silencing attenuates DDP resistance of NSCLC cells.

• METTL14 promotes the nature of pri-miR-19a by upregulating pri-miR-19a m6A level.

• miR-19a-5p targets RBM24, thus reducing the binding of RBM24 and AXIN1 and inhibiting AXIN1 transcription.

• METTL14 silencing in vivo enhances the suppressive role of DDP to tumor growth.

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Data availability

The authors confirm that the data supporting the findings of this study are available within the article.

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Funding

This research was supported by funding from the Science and Technology Foundation of Liaoning Province (2022-BS-122).

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Authors and Affiliations

  1. Department of Thoracic Surgery, First Affiliated Hospital of China Medical University, Liaoning Province, Shenyang, 110001, People’s Republic of China

    Shulei Gong & Mingrui Shao

  2. Department of Geriatric Surgery, First Affiliated Hospital of China Medical University, Liaoning Province, Shenyang, 110001, People’s Republic of China

    Shiyang Wang

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Contributions

All authors contributed to the study conception and design. Material preparation and data collection and analysis were performed by SLG and SYW. The first draft of the manuscript was written by MRS, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Mingrui Shao.

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This study was conducted with the approval of the ethics committee of the First Affiliated Hospital of China Medical University and in accordance with the Declaration of Helsinki. All participants signed written informed consent. Animal experimental protocols were approved by the animal ethics committee of the First Affiliated Hospital of China Medical University. The protocol was also conducted in compliance with the Guidelines for the Care and Use of Laboratory Animals provisions of administration and usage of laboratory animals [15].

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Gong, S., Wang, S. & Shao, M. Mechanism of METTL14-mediated m6A modification in non-small cell lung cancer cell resistance to cisplatin. J Mol Med 100, 1771–1785 (2022). https://doi.org/10.1007/s00109-022-02268-2

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