RNA m6A methylation regulates uveal melanoma cell proliferation, migration, and invasion by targeting c-Met
RNA m6A methylation regulates uveal melanoma cell proliferation, migration, and invasion by targeting c-Met
复制标题
RNA m(6)A 甲基化通过靶向 c-Met 调节葡萄膜黑色素瘤细胞增殖、迁移和侵袭
DOI：
10.1002/jcp.29608
复制
发表时间：
2020-02-04
影响因子：
5.6
通讯作者：
Yan, Dongsheng
中科院分区：
文献类型：
--
作者：
Luo, Guangying;Xu, Weiwei;Yan, Dongsheng
n -6-甲基腺苷（m(6)A）是一种新型的表转录组标记物，通过控制信使RNA代谢来调节多种生物过程。然而，尚不清楚m(6)A RNA甲基化是否影响葡萄膜黑色素瘤（UM）的发展。为了解决这个问题，我们探讨了它在UM中的作用和分子机制。最初，我们证明了在UM细胞系和临床标本中，全局RNA m(6)A甲基化水平显著升高。同时，我们发现UM细胞和标本中主要的m(6) a调节酶METTL3显著升高。随后，使用环亮氨酸（Cyc）或METTL3靶向小干扰RNA阻断UM细胞中的m(6)A甲基化。我们通过功能分析发现，Cyc或沉默METTL3通过细胞周期G1阻滞显著抑制UM细胞增殖和集落形成，以及迁移和侵袭。另一方面，过表达METTL3具有相反的作用。此外，生物信息学和甲基化RNA免疫沉淀-定量聚合酶链反应鉴定出c-Met是UM细胞中m(6) a甲基化的直接靶点。此外，western blot分析显示，Cyc或敲低METTL3可下调UM细胞中c-Met、p-Akt和细胞周期相关蛋白的水平。综上所述，我们的研究结果表明，mettl3介导的m(6)A RNA甲基化通过靶向c-Met调节UM细胞的增殖、迁移和侵袭。这种修饰在UM的发展中起着关键的致癌调节作用。
N-6-methyladenosine (m(6)A) is a novel epitranscriptomic marker that contributes to regulating diverse biological processes through controlling messenger RNA metabolism. However, it is unknown if m(6)A RNA methylation affects uveal melanoma (UM) development. To address this question, we probed its function and molecular mechanism in UM. Initially, we demonstrated that global RNA m(6)A methylation levels were dramatically elevated in both UM cell lines and clinical specimens. Meanwhile, we found that METTL3, a main m(6)A regulatory enzyme, was significantly increased in UM cells and specimens. Subsequently, cycloleucine (Cyc) or METTL3 targeted small interfering RNA was used to block m(6)A methylation in UM cells. We found that Cyc or silencing METTL3 significantly suppressed UM cell proliferation and colony formation through cell cycle G1 arrest, as well as migration and invasion by functional analysis. On the other hand, overexpression of METTL3 had the opposite effects. Furthermore, bioinformatics and methylated RNA immunoprecipitation-quantitative polymerase chain reaction identified c-Met as a direct target of m(6)A methylation in UM cells. In addition, western blot analysis showed that Cyc or knockdown of METTL3 downregulated c-Met, p-Akt, and cell cycle-related protein levels in UM cells. Taken together, our results demonstrate that METTL3-mediated m(6)A RNA methylation modulates UM cell proliferation, migration, and invasion by targeting c-Met. Such a modification acts as a critical oncogenic regulator in UM development.