m6A demethylase ALKBH5 inhibits tumor growth and metastasis by reducing YTHDFs-mediated YAP expression and inhibiting miR-107/LATS2-mediated YAP activity in NSCLC
m6A demethylase ALKBH5 inhibits tumor growth and metastasis by reducing YTHDFs-mediated YAP expression and inhibiting miR-107/LATS2-mediated YAP activity in NSCLC
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DOI：
10.1186/s12943-020-01161-1
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发表时间：
2020-02-27
期刊：
影响因子：
37.3
通讯作者：
Sun, Hongliang
中科院分区：
文献类型：
--
作者：
Jin, Dan;Guo, Jiwei;Sun, Hongliang
背景：被ALKBH5消除的mRNA甲基化在mRNA的生物发生、衰退和翻译控制中的重要性是一个新兴的研究热点。异位激活的YAP与许多人类癌症的发生有关。然而，ALKBH5调节YAP表达和活性以抑制NSCLC肿瘤生长和转移的机制尚不清楚。方法分析正常肺细胞和非小细胞肺癌细胞中蛋白质和转录产物的相互作用。用定量聚合酶链式反应和报告基因分析检测基因的表达。用免疫化学方法测定蛋白质水平。用免疫沉淀法分析核酸的相互作用和状态。通过标准的生化测试分析细胞行为。M(6)A修饰用MERIP分析。结果YAP的表达与ALKBH5的表达呈负相关，在NSCLC细胞的增殖、侵袭、迁移和EMT的调控中起相反的作用。ALKBH5减少了m(6)YAP的A修饰。YTHDF3结合YAP前mRNA依赖于m(6)A修饰。YTHDF1和YTHDF2以m(6)A非依赖的方式竞争性地与YTHDF3相互作用，调节YAP的表达。YTHDF2通过AGO2系统促进Yap mRNA的降解，而YTHDF1通过与eIF3a相互作用促进Yap mRNA的翻译，这两种活性都受m(6)A修饰的调节。此外，ALKBH5通过调节miR-107/LATS2轴以一种HUR依赖的方式降低YAP活性。此外，ALKBH5通过降低YAP的表达和活性来抑制体内肿瘤的生长和转移。结论m(6)A去甲基酶ALKBH5通过抑制YTHDFS介导的YAP表达和抑制miR-107/LATS2介导的YAP活性抑制NSCLC的肿瘤生长和转移。此外，有效抑制m(6)A修饰的ALKBH5可能构成一种潜在的肺癌治疗策略。
Background The importance of mRNA methylation erased by ALKBH5 in mRNA biogenesis, decay, and translation control is an emerging research focus. Ectopically activated YAP is associated with the development of many human cancers. However, the mechanism whereby ALKBH5 regulates YAP expression and activity to inhibit NSCLC tumor growth and metastasis is not clear. Methods Protein and transcript interactions were analyzed in normal lung cell and NSCLC cells. Gene expression was evaluated by qPCR and reporter assays. Protein levels were determined by immunochemical approaches. Nucleic acid interactions and status were analyzed by immunoprecipitation. Cell behavior was analyzed by standard biochemical tests. The m(6)A modification was analyzed by MeRIP. Results Our results show that YAP expression is negatively correlated with ALKBH5 expression and plays an opposite role in the regulation of cellular proliferation, invasion, migration, and EMT of NSCLC cells. ALKBH5 reduced m(6)A modification of YAP. YTHDF3 combined YAP pre-mRNA depending on m(6)A modification. YTHDF1 and YTHDF2 competitively interacted with YTHDF3 in an m(6)A-independent manner to regulate YAP expression. YTHDF2 facilitated YAP mRNA decay via the AGO2 system, whereas YTHDF1 promoted YAP mRNA translation by interacting with eIF3a; both these activities are regulated by m(6)A modification. Furthermore, ALKBH5 decreased YAP activity by regulating miR-107/LATS2 axis in an HuR-dependent manner. Further, ALKBH5 inhibited tumor growth and metastasis in vivo by reducing the expression and activity of YAP. Conclusions The presented findings suggest m(6)A demethylase ALKBH5 inhibits tumor growth and metastasis by reducing YTHDFs-mediated YAP expression and inhibiting miR-107/LATS2-mediated YAP activity in NSCLC. Moreover, effective inhibition of m(6)A modification of ALKBH5 might constitute a potential treatment strategy for lung cancer.