METTL3 介导的 mA mRNA 甲基化直接促进 YAP 翻译，并通过调节 MALAT1-miR-1914-3p-YAP 轴增加 YAP 活性，从而诱导 NSCLC 耐药和转移。

mA mRNA methylation initiated by METTL3 directly promotes YAP translation and increases YAP activity by regulating the MALAT1-miR-1914-3p-YAP axis to induce NSCLC drug resistance and metastasis.

机构信息

Clinical Medical Laboratory, Binzhou Medical University Hospital, Binzhou, 256603, People's Republic of China.

Cancer research institute, Binzhou Medical University Hospital, Binzhou, 256603, People's Republic of China.

出版信息

J Hematol Oncol. 2019 Dec 9;12(1):135. doi: 10.1186/s13045-019-0830-6.

DOI:10.1186/s13045-019-0830-6

PMID:31818312

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6902496/

Abstract

BACKGROUND

METTL3 is an RNA methyltransferase that mediates mA 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, mA modification was analyzed by MeRIP.

RESULTS

METTL3 increased the mA 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 mA 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 mA modification by METTL3 knockdown inhibits tumor growth and enhances sensitivity to DDP in vivo.

CONCLUSION

Results indicated that the mA 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.

摘要

背景

METTL3 是一种 RNA 甲基转移酶，介导 mA 修饰，并参与 mRNA 的生成、降解和翻译。然而，METTL3 通过调节 MALAT1-miR-1914-3p-YAP 轴活性来诱导 NSCLC 耐药和转移的生物力学机制尚不清楚。

方法

通过 qPCR 检测分析 mRNA 的表达。通过 Western blot 和免疫荧光染色分析蛋白水平。通过 CCK8 检测分析细胞增殖。通过划痕愈合和 Transwell 分析分别检测细胞迁移和侵袭。通过荧光素酶报告基因检测分析启动子活性和基因转录。最后，通过 MeRIP 分析 mA 修饰。

结果

METTL3 增加了 YAP 的 mA 修饰。METTL3、YTHDF3、YTHDF1 和 eIF3b 通过与翻译起始机制相互作用，直接促进 YAP 的翻译。此外，由于 METTL3 介导的更高水平的 mA 修饰，MALAT1 的 RNA 水平增加。同时，MALAT1 的稳定性通过 METTL3/YTHDF3 复合物增加。此外，MALAT1 作为一种竞争性内源性 RNA，通过 YAP 促进 NSCLC 的侵袭和转移，其作用是通过海绵吸附 miR-1914-3p。此外，METTL3 敲低减少 YAP 的 mA 修饰可抑制体内肿瘤生长并增强对 DDP 的敏感性。