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 Biology and Toxicology

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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
Cell Biology and Toxicology ( IF 5.3 ) Pub Date : 2022-08-15 , DOI: 10.1007/s10565-022-09751-z
Peng Ruan ₁ , Shujun Wang ₂ , Chaoyi Yang ₂ , Xiaohui Huang ₂ , Pengxing Sun ₂ , Aili Tan ₂

Affiliation

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.

中文翻译：

m6A mRNA甲基化通过PAPPA/IGFBP4轴调节ERK/NF-κB/AKT信号通路促进子宫内膜癌增殖和肿瘤形成

N6-甲基腺苷 (m ⁶ A) mRNA 甲基化被认为是参与疾病进展和癌变的基因调节机制。本论文旨在探讨m ⁶ A mRNA甲基化在子宫内膜癌中的具体机制。采用RT-qPCR来测试m ⁶ A甲基化与子宫内膜癌之间的临床相关性。通过生物信息学分析筛选子宫内膜癌相关基因，并利用SRAMP预测m ⁶ A靶点。通过Western blot检测和MeRIP-qPCR实验验证m ⁶ A甲基化对子宫内膜癌发生相关候选基因及信号通路的影响。m ⁶ A-seq、RT-qPCR 和多核糖体分析用于确认 m ⁶ A 甲基化调节相关基因和通路的机制。^{子宫内膜癌患者肿瘤组织中m 6} A甲基化、METTL3和IGFBP4水平降低，SRAMP分析证实IGFBP4和PAPPA存在m ⁶ A甲基化位点。m6A甲基化减少促进子宫内膜癌细胞进展和体内肿瘤形成。m ⁶子宫内膜癌细胞中 RNA 的甲基化直接调节 IGFBP4 和 PAPPA 的表达。m ⁶ A 甲基化调节 PAPPA/IGFBP4 轴，从而通过 NF-κB 和 ERK 信号通路影响子宫内膜癌。子宫内膜癌细胞中 PAPPA 的敲低或 IGFBP4 的过度表达部分减缓了 m ⁶ A 甲基化减少引起的疾病进展。这项研究表明，m ⁶ A mRNA 甲基化通过 PAPPA/IGFBP4 轴调节 ERK/NF-κB/AKT 信号通路，从而诱导子宫内膜癌中的细胞增殖和肿瘤形成。