DNMT1-MicroRNA126 Epigenetic Circuit Contributes to Esophageal Squamous Cell Carcinoma Growth via ADAM9-EGFR-AKT Signaling
DNMT1-MicroRNA126 Epigenetic Circuit Contributes to Esophageal Squamous Cell Carcinoma Growth via ADAM9-EGFR-AKT Signaling
复制标题
DNMT1-MicroRNA126 表观遗传电路通过 ADAM9-EGFR-AKT 信号传导促进食管鳞状细胞癌生长。
DOI：
10.1158/1078-0432.ccr-14-1740
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发表时间：
2015-02-15
影响因子：
11.5
通讯作者：
Chu, Yiwei
中科院分区：
文献类型：
--
作者：
Liu, Ronghua;Gu, Jie;Chu, Yiwei
目的：MicroRNA（miRNA）参与表观遗传调控并受其控制，从而形成相互的调控回路。本研究利用基于下一代测序 (NGS) 的 miRNA 分析，旨在发现食管鳞状细胞癌 (ESCC) 特异性 miRNA 和 miRNA 相关的表观遗传调节。 实验设计：为四对 ESCC 组织和邻近正常组织生成基于 NGS 的 miRNA 谱。原位杂交用于评估 miRNA 表达及其与预后的相关性。使用亚硫酸氢盐基因组测序鉴定了 miRNA 相关的 DNA 甲基化，并使用 RNA 干扰研究了 DNA 甲基转移酶 1 (DNMT1) 的作用。通过 mRNA 测序筛选 miRNA 靶标，并在体外和体内进行功能验证。结果：基于 NGS 的 miRNA 分析鉴定出 ESCC 中 78 个差异表达的 miRNA。其中，microRNA126-3p（miR-126）显着下调，其下调与ESCC预后不良相关。 miR-126 的下调是由于其宿主基因 Egfl7 的启动子高甲基化。 DNMT1 在 ESCC 中异常上调，并导致 Egfl7 的高甲基化。有趣的是，DNMT1 被 miR-126 的过度表达所抑制，表明调节反馈回路的存在。 ADAM9 被确定为 miR-126 的关键靶标。 miR-126 的异位表达或 ADAM9 的沉默通过抑制表皮生长因子受体 -AKT 信号传导来减少 ESCC 细胞的增殖和迁移。结论：我们的结果表明 miR-126 是 ESCC 的潜在预后指标，并表明一种新的“DNMT1-miR-126 表观遗传回路”参与 ESCC 进展。因此，基于 miR-126 的表观遗传调节可能为抗肿瘤治疗的新方法提供基本原理。 (C) 2014 年 AACR。
Purpose: MicroRNAs (miRNA) are involved in and are controlled by epigenetic regulation, and thereby form a reciprocal regulatory circuit. Using next-generation sequencing (NGS)based miRNA profiling, this study aimed to discover esophageal squamous cell carcinoma (ESCC)-specific miRNAs and miRNArelated epigenetic modulations.Experimental Design: NGS-based miRNA profiles were generated for four pairs of ESCC tissues and adjacent normal tissues. In situ hybridization was used to assess miRNA expression and its correlation with prognosis. miRNA-related DNA methylations were identified using bisulfite genomic sequencing, and the role of DNA methyltransferase 1 (DNMT1) was investigated using RNA interference. miRNA targets were screened by mRNA sequencing, and functional validation was performed in vitro and in vivo.Results: NGS-based miRNA profiling identified 78 differentially expressed miRNAs in ESCC. Among them, microRNA126-3p (miR-126) was significantly downregulated, and its downregulation correlated with poor ESCC prognosis. Downregulation of miR-126 was due to promoter hypermethylation of its host gene, Egfl7. DNMT1 was aberrantly upregulated in ESCC and responsible for the hypermethylation of Egfl7. Intriguingly, DNMT1was suppressed by overexpression of miR-126, indicating the existence of a regulatory feedback circuit. ADAM9 was identified as a key target of miR-126. Ectopic expression of miR-126 or silencing of ADAM9 reduced ESCC cell proliferation and migration by inhibiting epidermal growth factor receptor-AKT signaling.Conclusions: Our results indicate that miR-126 is a potential prognostic indicator for ESCC and suggest that a novel "DNMT1-miR-126 epigenetic circuit" is involved in ESCC progression. Consequently, miR-126-based epigenetic modulations may provide a basic rationale for new approaches to antitumor therapeutics. (C) 2014 AACR.