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

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. 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.