Department of Thyroid and Breast Surgery, The Seventh Affiliated Hospital, Southern Medical University, Foshan, 528200, China.
Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China.
J Exp Clin Cancer Res. 2023 Jan 7;42(1):10. doi: 10.1186/s13046-022-02581-1.
Posttranscriptional modification of tumor-associated factors plays a pivotal role in breast cancer progression. However, the underlying mechanism remains unknown. M6A modifications in cancer cells are dynamic and reversible and have been found to impact tumor initiation and progression through various mechanisms. In this study, we explored the regulatory mechanism of breast cancer cell proliferation and metabolism through m6A methylation in the Hippo pathway. METHODS: A combination of MeRIP-seq, RNA-seq and metabolomics-seq was utilized to reveal a map of m6A modifications in breast cancer tissues and cells. We conducted RNA pull-down assays, RIP-qPCR, MeRIP-qPCR, and RNA stability analysis to identify the relationship between m6A proteins and LATS1 in m6A regulation in breast cancer cells. The expression and biological functions of m6A proteins were confirmed in breast cancer cells in vitro and in vivo. Furthermore, we investigated the phosphorylation levels and localization of YAP/TAZ to reveal that the activity of the Hippo pathway was affected by m6A regulation of LATS1 in breast cancer cells. RESULTS: We demonstrated that m6A regulation plays an important role in proliferation and glycolytic metabolism in breast cancer through the Hippo pathway factor, LATS1. METTL3 was identified as the m6A writer, with YTHDF2 as the reader protein of LATS1 mRNA, which plays a positive role in promoting both tumorigenesis and glycolysis in breast cancer. High levels of m6A modification were induced by METTL3 in LATS1 mRNA. YTHDF2 identified m6A sites in LATS1 mRNA and reduced its stability. Knockout of the protein expression of METTL3 or YTHDF2 increased the expression of LATS1 mRNA and suppressed breast cancer tumorigenesis by activating YAP/TAZ in the Hippo pathway.
In summary, we discovered that the METTL3-LATS1-YTHDF2 pathway plays an important role in the progression of breast cancer by activating YAP/TAZ in the Hippo pathway.
肿瘤相关因子的转录后修饰在乳腺癌的进展中起着关键作用。然而，其潜在机制尚不清楚。癌细胞中的 m6A 修饰是动态和可逆的，已被发现通过多种机制影响肿瘤的起始和进展。在这项研究中，我们通过 Hippo 通路中的 m6A 甲基化探索了乳腺癌细胞增殖和代谢的调节机制。
采用 MeRIP-seq、RNA-seq 和代谢组学-seq 联合方法，揭示乳腺癌组织和细胞中 m6A 修饰图谱。我们进行了 RNA 下拉实验、RIP-qPCR、MeRIP-qPCR 和 RNA 稳定性分析，以鉴定 m6A 蛋白与 LATS1 在乳腺癌细胞中 m6A 调控中的关系。在体外和体内证实了 m6A 蛋白在乳腺癌细胞中的表达和生物学功能。此外，我们还研究了 YAP/TAZ 的磷酸化水平和定位，揭示了 Hippo 通路的活性受到乳腺癌细胞中 LATS1 的 m6A 调控的影响。
我们证明了 m6A 调节通过 Hippo 通路因子 LATS1 在乳腺癌中对增殖和糖酵解代谢起重要作用。鉴定出 METTL3 是 m6A 书写酶，YTHDF2 是 LATS1 mRNA 的阅读器蛋白，它在促进乳腺癌的肿瘤发生和糖酵解中起积极作用。METTL3 在 LATS1 mRNA 上诱导高水平的 m6A 修饰。YTHDF2 在 LATS1 mRNA 上识别 m6A 位点并降低其稳定性。敲除 METTL3 或 YTHDF2 的蛋白表达会增加 LATS1 mRNA 的表达，并通过 Hippo 通路中的 YAP/TAZ 激活抑制乳腺癌肿瘤发生。
总之，我们发现 METTL3-LATS1-YTHDF2 通路通过 Hippo 通路中的 YAP/TAZ 激活在乳腺癌的进展中起着重要作用。
