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N6-methyladenosine-induced miR-182-5p promotes multiple myeloma tumorigenesis by regulating CAMK2N1

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Abstract

Methyltransferase like 3 (METTL3) has been reported to promote tumorigenesis of multiple myeloma (MM), however, the molecular mechanism still needs further research. The N6-methyladenosine (m6A) level in tissues or cells was measured by m6A kit and dot blot assay. The mRNA and protein expression were detected by quantitative real-time PCR (RT-qPCR) and Western blot, respectively. The cell counting kit-8 and colony formation assay were used to detect the cell proliferation. Coimmunoprecipitation (Co-IP) experiment verified the binding of two proteins. The luciferase reporter experiment demonstrated the targeted binding of miR-182-5p and CaMKII inhibitor 1 (CAMK2N1). More importantly, tumor growth was measured in xenograft mice. Our data showed that the expression of METTL3 was significantly increased in MM patients’ samples and MM cells. METTL3 overexpression promoted MM cells proliferation, and METTL3 knockdown inhibited MM cells proliferation. Mechanically, METTL3-dependent m6A participated in DiGeorge syndrome critical region 8 (DGCR8)-mediated maturation of pri-miR-182. Upregulation of miR-182-5p further enhanced the promoting proliferation effect of METTL3 overexpression on MM cells. Moreover, the luciferase reporter gene experiment proved that miR-182-5p targetedly regulated CAMK2N1 expression. Xenograft tumor in nude mice further verified that METTL3 promoted MM tumor growth through miR-182/CAMK2N1 signal axis. In summary, the METTL3/miR-182-5p/CAMK2N1 axis plays an important role in MM tumorigenesis, which may provide a new target for MM therapy.

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Data availability

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

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Funding

This research was supported by Anhui Provincial Department of Education (2022AH051178), the Anhui Natural Science Foundation of China (2008085MH296), the Anhui Medical University Natural Science Foundation (2020xkj186).

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  1. Department of Hematology, the First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, Hefei, 230022, Anhui, China

    Jing Bao, Tingting Xu, Wanjie Wang, Han Xu, Xiaowen Chen & Ruixiang Xia

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Contributions

JB, TTX, WJW, HX and XWC collected clinical data. JB and TTX carried out the experiments and analyzed this data. The manuscript was written and interpreted by JB. Study design, experiments and manuscript review were handled by RXX. The final manuscript was approved by all authors.

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Correspondence to Ruixiang Xia.

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For patients: This project was approved by the Research Ethics Committee of Anhui Medical University (No. 20200040). All participants in this project signed the informed consent form.

For animals: The animal experiments were carried out with the approval of the ethics committee of Anhui Medical University (No. 20190657).

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Bao, J., Xu, T., Wang, W. et al. N6-methyladenosine-induced miR-182-5p promotes multiple myeloma tumorigenesis by regulating CAMK2N1. Mol Cell Biochem (2024). https://doi.org/10.1007/s11010-023-04906-w

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