PDF(Pubmed)
Abstract：
RNA 5-methylcytosine (m5C) is a widespread post-transcriptional modification involved in diverse biological processes through controlling RNA metabolism. However, its roles in uveal melanoma (UM) remain unknown. Here, we describe the biological roles and regulatory mechanisms of RNA m5C in UM. Initially, we identified significantly elevated global RNA m5C levels in both UM cells and tissue specimens using ELISA assay and dot blot analysis. Meanwhile, NOP2/Sun RNA methyltransferase family member 2 (NSUN2) was upregulated in both types of these samples, whereas NSUN2 knockdown significantly decreased RNA m5C level. Such declines inhibited UM cell migration and suppressed cell proliferation through cell cycle G1 arrest. Furthermore, bioinformatic analyses, m5C-RIP-qPCR, and luciferase assay identified β-Catenin (CTNNB1) as a direct target of NSUN2-mediated m5C modification in UM cells. Additionally, overexpression of miR-124a in UM cells diminished NSUN2 expression levels indicating that it is an upstream regulator of this response. Our study suggests that NSUN2-mediated RNA m5C methylation provides a potential novel target to improve the therapeutic management of UM pathogenesis.
摘要：
RNA5-甲基胞嘧啶（m5C）是一种广泛的转录后修饰，通过控制RNA代谢参与各种生物过程。然而,其在葡萄膜黑色素瘤（UM）中的作用尚不清楚。这里,我们描述了RNAm5C在UM中的生物学作用和调节机制。最初，我们使用ELISA分析和斑点印迹分析在UM细胞和组织标本中鉴定出全局RNAm5C水平显著升高.同时,NOP2/SunRNA甲基转移酶家族成员2（NSUN2）在这两种类型的样品中上调，而NSUN2敲低显著降低m5CRNA水平。这种下降抑制UM细胞迁移并通过细胞周期G1阻滞抑制细胞增殖。此外，生物信息学分析，m5C-RIP-qPCR，荧光素酶分析确定β-连环蛋白（CTNNB1）是UM细胞中NSUN2介导的m5C修饰的直接靶标。此外,miR-124a在UM细胞中的过表达降低了NSUN2表达水平,表明它是该反应的上游调节因子。我们的研究表明，NSUN2介导的RNAm5C甲基化为改善UM发病机制的治疗管理提供了一个潜在的新靶标。
