WTAP facilitates progression of hepatocellular carcinoma via m6A-HuR-dependent epigenetic silencing of ETS1
WTAP facilitates progression of hepatocellular carcinoma via m6A-HuR-dependent epigenetic silencing of ETS1
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DOI：
10.1186/s12943-019-1053-8
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发表时间：
2019-08-22
期刊：
影响因子：
37.3
通讯作者：
Zheng, Shusen
中科院分区：
文献类型：
--
作者：
Chen, Yunhao;Peng, Chuanhui;Zheng, Shusen
n6 -甲基腺苷（m6A）甲基化是一种众所周知的具有新的表观遗传功能的修饰，已被报道参与肝细胞癌（HCC）的肿瘤发生，为该疾病的分子发病机制提供了新的见解。然而，作为m6A甲基化的关键成分，Wilms肿瘤1相关蛋白（WTAP）在HCC中的研究尚不充分。本研究探讨WTAP在肝癌中的生物学作用及其潜在机制。方法利用组织微阵列和肿瘤基因组图谱（TCGA）数据集检测WTAP的表达及其与临床病理特征的相关性。通过细胞增殖实验、集落形成实验、Edu实验和皮下异种移植实验，明确了WTAP对肝癌细胞的影响。然后，我们利用RNA测序结合基因表达综合（GEO）数据筛选WTAP候选靶点。最后，我们通过m6A点印迹法、甲基化RNA免疫沉淀法（MeRIP）、双荧光素酶报告基因法、RNA免疫沉淀法（RIP）和染色质免疫沉淀法（ChIP）研究了WTAP在HCC中的调控机制。结果我们发现WTAP在HCC中高表达，提示预后不良，并且WTAP表达可作为HCC生存的独立预测因子。在功能上，WTAP在体外和体内均能促进HCC细胞的增殖能力和肿瘤生长。此外，ETS原癌基因1 （ETS1）被确定为WTAP的下游效应因子。WTAP调控的m6A修饰导致ETS1转录后抑制，这表明HuR抗原R （HuR）是RNA稳定剂。然后发现ETS1可以抑制HCC的进展，并可以挽救WTAP缺乏引起的表型。此外，WTAP通过ETS1介导的p21/p27依赖模式调节HCC细胞的G2/M期。结论WTAP在HCC中显著上调，促进肝癌的发展。wtap引导的m6A修饰通过hr - ets1 -p21/p27轴促进HCC的进展。我们的研究首次报道了WTAP介导的m6A甲基化在HCC的发生中起着至关重要的作用，并强调了WTAP作为HCC治疗的潜在治疗靶点。
BackgroundN6-methyladenosine (m6A) methylation, a well-known modification with new epigenetic functions, has been reported to participate in the tumorigenesis of hepatocellular carcinoma (HCC), providing novel insights into the molecular pathogenesis of this disease. However, as the key component of m6A methylation, Wilms tumor 1-associated protein (WTAP) has not been well studied in HCC. Here we investigated the biological role and underlying mechanism of WTAP in liver cancer.MethodsWe determined the expression of WTAP and its correlation with clinicopathological features using tissue microarrays and the Cancer Genome Atlas (TCGA) dataset. And we clarified the effects of WTAP on HCC cells using cell proliferation assay, colony formation, Edu assay and subcutaneous xenograft experiments. We then applied RNA sequencing combined with gene expression omnibus (GEO) data to screen candidate targets of WTAP. Finally, we investigated the regulatory mechanism of WTAP in HCC by m6A dot blot assay, methylated RNA immunoprecipitation (MeRIP) assay, dual luciferase reporter assay, RNA immunoprecipitation (RIP) assay and Chromatin immunoprecipitation (ChIP) assay.ResultsWe demonstrated that WTAP was highly expressed in HCC which indicated the poor prognosis, and that WTAP expression served as an independent predictor of HCC survival. Functionally, WTAP promoted the proliferation capability and tumor growth of HCC cells in vitro and in vivo. Furthermore, ETS proto-oncogene 1 (ETS1) was identified as the downstream effector of WTAP. The m6A modification regulated by WTAP led to post-transcriptional suppression of ETS1, with the implication of Hu-Antigen R (HuR) as an RNA stabilizer. Then ETS1 was found to inhibit the progression of HCC and could rescue the phenotype induced by WTAP deficiency. Moreover, WTAP modulated the G2/M phase of HCC cells through a p21/p27-dependent pattern mediated by ETS1.ConclusionWe have identified that WTAP is significantly up-regulated in HCC and promotes liver cancer development. WTAP-guided m6A modification contributes to the progression of HCC via the HuR-ETS1-p21/p27 axis. Our study is the first to report that WTAP-mediated m6A methylation has a crucial role in HCC oncogenesis, and highlights WTAP as a potential therapeutic target of HCC treatment.