BACKGROUND METTL3 is an RNA methyltransferase that mediates m6A modification and is implicated in mRNA biogenesis, decay, and translation. However, the biomechanism through which METTL3 regulates MALAT1-miR-1914-3p-YAP axis activity to induce NSCLC drug resistance and metastasis is not very clear. METHODS The expression of mRNA was analyzed by qPCR assays. Protein levels were analyzed by western blotting and immunofluorescent staining. Cellular proliferation was detected by CCK8 assays. Cell migration and invasion were analyzed by wound healing and transwell assays, respectively. Promoter activities and gene transcription were analyzed by luciferase reporter assays. Finally, m6A modification was analyzed by MeRIP. RESULTS METTL3 increased the m6A modification of YAP. METTL3, YTHDF3, YTHDF1, and eIF3b directly promoted YAP translation through an interaction with the translation initiation machinery. Moreover, the RNA level of MALAT1 was increased due to a higher level of m6A modification mediated by METTL3. Meanwhile, the stability of MALAT1 was increased by METTL3/YTHDF3 complex. Additionally, MALAT1 functions as a competing endogenous RNA that sponges miR-1914-3p to promote the invasion and metastasis of NSCLC via YAP. Furthermore, the reduction of YAP m6A modification by METTL3 knockdown inhibits tumor growth and enhances sensitivity to DDP in vivo. CONCLUSION Results indicated that the m6A mRNA methylation initiated by METTL3 promotes YAP mRNA translation via recruiting YTHDF1/3 and eIF3b to the translation initiation complex and increases YAP mRNA stability through regulating the MALAT1-miR-1914-3p-YAP axis. The increased YAP expression and activity induce NSCLC drug resistance and metastasis.

中文翻译：

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METTL3 启动的 m6A mRNA 甲基化通过调节 MALAT1-miR-1914-3p-YAP 轴直接促进 YAP 翻译并增加 YAP 活性，从而诱导 NSCLC 耐药和转移。

背景 METTL3 是一种 RNA 甲基转移酶，可介导 m6A 修饰并参与 mRNA 生物发生、衰变和翻译。然而，METTL3通过调控MALAT1-miR-1914-3p-YAP轴活性诱导NSCLC耐药和转移的生物机制尚不十分清楚。方法 通过 qPCR 检测分析 mRNA 的表达。通过蛋白质印迹和免疫荧光染色分析蛋白质水平。通过CCK8测定法检测细胞增殖。细胞迁移和侵袭分别通过伤口愈合和 transwell 测定进行分析。通过荧光素酶报告基因测定分析启动子活性和基因转录。最后，通过 MeRIP 分析 m6A 修饰。结果 METTL3 增加了 YAP 的 m6A 修饰。METTL3、YTHDF3、YTHDF1、eIF3b 通过与翻译启动机制的交互直接促进 YAP 翻译。此外，由于 METTL3 介导的更高水平的 m6A 修饰，MALAT1 的 RNA 水平增加。同时，METTL3/YTHDF3 复合物增加了 MALAT1 的稳定性。此外，MALAT1 作为一种竞争性内源性 RNA，通过 YAP 吸收 miR-1914-3p 促进 NSCLC 的侵袭和转移。此外，通过 METTL3 敲低减少 YAP m6A 修饰可抑制肿瘤生长并增强体内对 DDP 的敏感性。结论 结果表明，METTL3 启动的 m6A mRNA 甲基化通过将 YTHDF1/3 和 eIF3b 募集到翻译起始复合物来促进 YAP mRNA 翻译，并通过调节 MALAT1-miR-1914-3p-YAP 轴增加 YAP mRNA 的稳定性。