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m6A mRNA methylation regulates the ERK/NF-κB/AKT signaling pathway through the PAPPA/IGFBP4 axis to promote proliferation and tumor formation in endometrial cancer

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Abstract

N6-methyladenosine (m6A) mRNA methylation has been considered a gene modulatory mechanism involved in disease progression and carcinogenesis. Herein, we aimed to explore the specific mechanism of m6A mRNA methylation in endometrial cancer. RT-qPCR was implemented to test the clinical correlation between m6A methylation and endometrial cancer. Bioinformatics analysis was performed to screen the genes related to endometrial cancer, and SRAMP was utilized for the prediction of m6A targets. Western blot assay and MeRIP-qPCR experiments were conducted to verify the effect of m6A methylation on the candidate genes and the signaling pathways involved in the occurrence of endometrial cancer. m6A-seq, RT-qPCR, and polysome profiling were used to confirm the mechanisms of m6A methylation in modulating related genes and pathways. The levels of m6A methylation, METTL3, and IGFBP4 were reduced in tumor tissues of patients with endometrial cancer, and SRAMP analysis confirmed that IGFBP4 and PAPPA had m6A methylation sites. Reduced m6A methylation promoted endometrial cancer cell progression and tumor formation in vivo. m6A methylation of RNA in endometrial cancer cells directly modulated IGFBP4 and PAPPA expression. m6A methylation regulated the PAPPA/IGFBP4 axis, thereby influencing endometrial cancer through the NF-κB and ERK signaling pathways. Knockdown of PAPPA or overexpression of IGFBP4 in endometrial cancer cells partially reduced disease progression caused by reduced m6A methylation. This research suggests that m6A mRNA methylation modulates the ERK/NF-κB/AKT signaling pathway through the PAPPA/IGFBP4 axis to induce cell proliferation and tumor formation in endometrial cancer.

Graphical abstract

1. METTL3 expressed modestly and m6A methylation of IGFBP4 and PAPPA mRNAs decreased in endometrial cancer;

2. YTHDF1-mediated IGFBP4 translation was reduced in HEC-1-A and AN3CA cells, and YTHDF2-mediated PAPPA mRNA degradation was blunted but its protein expression increased;

3. Increased PAPPA and reduced IGFBP4 activated IGF1-induced ERK, AKT, and NF-κB pathways by binding IGFR, thereby promoting cancer cell malignancy.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

Code availability

Not applicable.

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Funding

Thanks for the grants from the National Natural Science Foundation of China (No. 81801421).

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Authors and Affiliations

  1. Department of Oncology, Renmin Hospital of Wuhan University, Wuhan, Hubei, 430060, People’s Republic of China

    Peng Ruan

  2. Department of Obstetrics & Gynecology, Renmin Hospital of Wuhan University, No. 99 Zhangzhidong Road, Wuchang District, Wuhan, Hubei, 430060, People’s Republic of China

    Shujun Wang, Chaoyi Yang, Xiaohui Huang, Pengxing Sun & Aili Tan

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Contributions

RP and TAL conceived the ideas. RP, TAL, and WSJ designed the experiments. RP, YCY, and HXH performed the experiments. RP and SPX analyzed the data. RP and TAL provided critical materials. RP, TAL, and WSJ wrote the manuscript. LHX supervised the study. All the authors have read and approved the final version for publication.

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Correspondence to Aili Tan.

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The clinical and animal experiments were ratified by the ethics committee of of Renmin Hospital of Wuhan University and the relevant studies were completed within the scope of ethical regulations. The clinical experiments conformed to the Declaration of Helsinki. All subjects had provided their written informed consent. All animal experiments were in strict accordance with the rules and regulations and operating norms for the management of experimental animals and the relevant ethical requirements of experimental animals.

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Informed consent was obtained from all individual participants included in the study.

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Ruan, P., Wang, S., Yang, C. et al. m6A mRNA methylation regulates the ERK/NF-κB/AKT signaling pathway through the PAPPA/IGFBP4 axis to promote proliferation and tumor formation in endometrial cancer. Cell Biol Toxicol 39, 1611–1626 (2023). https://doi.org/10.1007/s10565-022-09751-z

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