m6A modification suppresses ocular melanoma through modulating HINT2 mRNA translation
m6A modification suppresses ocular melanoma through modulating HINT2 mRNA translation
复制标题
m(6)A 修饰通过调节 HINT2 mRNA 翻译来抑制眼部黑色素瘤。
DOI：
10.1186/s12943-019-1088-x
复制
发表时间：
2019-11-14
期刊：
影响因子：
37.3
通讯作者：
Fan, Xianqun
中科院分区：
文献类型：
--
作者：
Jia, Ruobing;Chai, Peiwei;Fan, Xianqun
背景N-6-甲基腺苷（m（6）A）RNA的动态修饰由N-6-甲基转移酶和脱甲基酶产生和清除，调节基因表达、选择性剪接和细胞命运。眼黑色素瘤，包括葡萄膜黑色素瘤（UM）和结膜黑色素瘤（CM），是成人中最常见的原发性眼肿瘤，也是第二常见的黑色素瘤。然而，m（6）A修饰在眼黑色素瘤中的功能作用仍不清楚。方法采用m（6）A检测和生存分析来探讨总体m（6）A水平降低，提示眼黑色素瘤晚期和预后不良。miCLIP-seq、RNA-seq和Label-free MS数据的多组学分析显示，m（6）A RNA修饰在转录后促进HINT 2表达。RNA免疫沉淀（RIP）-qPCR和双荧光素酶测定显示HINT 2 mRNA与YTHDF 1特异性相互作用。此外，多核糖体分析表明，在具有较高m（6）A甲基化的眼黑色素瘤细胞中，翻译池中HINT 2 mRNA的量更大。结果在这里，我们发现RNA甲基化显著抑制UM和CM的进展。眼黑色素瘤样本显示m（6）A水平降低，提示预后不良。整体m（6）A修饰的变化与体外和体内肿瘤进展高度相关。从机制上讲，YTHDF 1促进了甲基化HINT 2 mRNA的翻译，HINT 2 mRNA是眼黑色素瘤中的肿瘤抑制因子。结论我们的工作揭示了m（6）A甲基化在眼黑色素瘤中的重要作用，并为理解m（6）A修饰提供了额外的见解。
Background Dynamic N-6-methyladenosine (m(6)A) RNA modification generated and erased by N-6-methyltransferases and demethylases regulates gene expression, alternative splicing and cell fate. Ocular melanoma, comprising uveal melanoma (UM) and conjunctival melanoma (CM), is the most common primary eye tumor in adults and the 2nd most common melanoma. However, the functional role of m(6)A modification in ocular melanoma remains unclear. Methods m(6)A assays and survival analysis were used to explore decreased global m(6)A levels, indicating a late stage of ocular melanoma and a poor prognosis. Multiomic analysis of miCLIP-seq, RNA-seq and Label-free MS data revealed that m(6)A RNA modification posttranscriptionally promoted HINT2 expression. RNA immunoprecipitation (RIP)-qPCR and dual luciferase assays revealed that HINT2 mRNA specifically interacted with YTHDF1. Furthermore, polysome profiling analysis indicated a greater amount of HINT2 mRNA in the translation pool in ocular melanoma cells with higher m(6)A methylation. Results Here, we show that RNA methylation significantly inhibits the progression of UM and CM. Ocular melanoma samples showed decreased m(6)A levels, indicating a poor prognosis. Changes in global m(6)A modification were highly associated with tumor progression in vitro and in vivo. Mechanistically, YTHDF1 promoted the translation of methylated HINT2 mRNA, a tumor suppressor in ocular melanoma. Conclusions Our work uncovers a critical function for m(6)A methylation in ocular melanoma and provides additional insight into the understanding of m(6)A modification.