m6A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma
m6A mRNA methylation regulates CTNNB1 to promote the proliferation of hepatoblastoma
复制标题
m(6)A mRNA甲基化调控CTNNB1促进肝母细胞瘤增殖
DOI：
10.1186/s12943-019-1119-7
复制
发表时间：
2019-12-23
期刊：
影响因子：
37.3
通讯作者：
Pan, Qiuhui
中科院分区：
文献类型：
--
作者：
Liu, Li;Wang, Jing;Pan, Qiuhui
背景N-6-甲基腺苷（m（6）A）修饰参与了许多生物学过程。它对信使RNA（mRNA）的稳定性、剪接和翻译的调节很重要。然而，它在癌症中的作用尚未详细研究。本文研究了m（6）A修饰在肝母细胞瘤（HB）中的生物学作用及其机制。方法采用逆转录实时定量PCR（RT-qPCR）和Western blotting方法检测m6 A相关因子的表达。并通过细胞增殖实验、集落形成实验、凋亡实验等研究了这些因子对HB细胞的影响。然后研究甲基转移酶样蛋白13（methyltransferase-like 13，简称L3）的表达及其与临床病理特征的关系，并通过异种移植实验检测L3在体内的作用。使用m（6）A-Seq分析肝母细胞瘤肿瘤组织和正常组织中m（6）A全转录组。最后，通过甲基化RNA免疫沉淀（MeRIP）试验、RNA残留试验来研究MEETL 3对HB中靶CTNNB 1的调节机制。结果在本研究中我们发现肝母细胞瘤中m（6）A修饰增多，而胃L3是参与异常m（6）A修饰的主要因素。我们还分析了肝母细胞瘤肿瘤组织和正常组织中整个转录组的m（6）A。我们的研究结果表明，m（6）A在肝母细胞瘤中高度表达。此外，m（6）A不仅在终止密码子周围富集，而且在编码序列（CDS）区域周围富集。基因本体分析表明m（6）A mRNA甲基化对Wnt/beta-catenin通路的调控有重要作用。减少m（6）A甲基化可导致CTNNB 1的表达和稳定性降低。结论肝母细胞瘤中m（6）A mRNA甲基化水平升高是一种致癌机制，胃L3在HB中表达上调，促进HB的发生发展。并鉴定CTNNB 1作为HB中胃L3引导的m（6）A修饰的调节剂。
Background N-6-Methyladenosine (m(6)A) modification has been implicated in many biological processes. It is important for the regulation of messenger RNA (mRNA) stability, splicing, and translation. However, its role in cancer has not been studied in detail. Here we investigated the biological role and underlying mechanism of m(6)A modification in hepatoblastoma (HB). Methods We used Reverse transcription quantitative real-time PCR (RT-qPCR) and Western blotting to determine the expression of m(6)A related factors. And we clarified the effects of these factors on HB cells using cell proliferation assay, colony formation, apoptotic assay. Then we investigated of methyltransferase-like 13 (METTL3) and its correlation with clinicopathological features and used xenograft experiment to check METTL3 effect in vivo. m(6)A-Seq was used to profiled m(6)A transcriptome-wide in hepatoblastoma tumor tissue and normal tissue. Finally, methylated RNA immunoprecipitation (MeRIP) assay, RNA remaining assay to perform the regulator mechanism of MEETL3 on the target CTNNB1 in HB. Results In this research, we discovered that m(6)A modifications are increased in hepatoblastoma, and METTL3 is the main factor involved with aberrant m(6)A modification. We also profiled m(6)A across the whole transcriptome in hepatoblastoma tumor tissues and normal tissues. Our findings suggest that m(6)A is highly expressed in hepatoblastoma tumors. Also, m(6)A is enriched not only around the stop codon, but also around the coding sequence (CDS) region. Gene ontology analysis indicates that m(6)A mRNA methylation contributes significantly to regulate the Wnt/beta-catenin pathway. Reduced m(6)A methylation can lead to a decrease in expression and stability of the CTNNB1. Conclusion Overall our findings suggest enhanced m(6)A mRNA methylation as an oncogenic mechanism in hepatoblastoma, METTL3 is significantly up-regulated in HB and promotes HB development. And identify CTNNB1 as a regulator of METTL3 guided m(6)A modification in HB.