胃癌中 EPB41L3 的表达、DNA 甲基化模式和转录因子:对 262 例病例的研究。

IF 8.2 2区 生物学 Q1 CELL BIOLOGY
Mengdi Cai, Haonan Guo, Dong Wang, Tie Zhao, Xiao Liang, Jiaqi Li, XiaoBo Cui, Songbin Fu, Jingcui Yu
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引用次数: 0

摘要

目的:DNA 甲基化会使肿瘤抑制基因失活并促进肿瘤发生。此前,我们发现了包含红细胞膜蛋白带 4.1-like 3(EPB41L3)基因的 18 号染色体缺失,该基因是胃癌(GC)中肺腺癌-1(DAL-1)不同表达的抑癌基因的祖先:我们目前的研究旨在阐明 EPB41L3 在 GC 中的表达和甲基化情况,识别调控转录因子,并确定受影响的下游通路。免疫组化结果表明,DAL-1在GC组织中的表达明显降低,其下调可作为独立的预后标志物:结果:对70对GC患者组织进行的高通量亚硫酸氢盐测序显示,EPB41L3启动子中非CpGs的甲基化程度较高与肿瘤恶性程度和组织分级较高有关。这种高甲基化被证明会降低 DAL-1 的表达,从而导致 GC 表型的恶性化。研究发现,DNA甲基转移酶抑制剂5-aza-2'-脱氧胞苷(5-aza-CdR)可部分恢复DAL-1的表达。此外,通过染色体免疫沉淀(ChIP)-qPCR和荧光素酶报告实验,确定了转录因子CDC5L与EPB41L3启动子上游区域的直接结合。免疫组化证实了 CDC5L 和 DAL-1 蛋白水平之间的正相关性。随后的RNA-seq分析显示,DAL-1对细胞外基质和空间相关通路有显著影响。5-Aza-CdR处理后的GC细胞RNA-seq和GC组织的单细胞RNA-seq数据证实,EPB41L3去甲基化或过表达后,GC上皮细胞中关键的肿瘤抑制因子AREG和COL17A1上调:总之,这项研究阐明了 EPB41L3 的非 CpG 甲基化与 GC 进展之间的关联,并确定了参与其中的关键转录因子和下游分子。这些发现加深了我们对 EPB41L3 在胃癌中作用的理解,为今后的研究和潜在的临床应用提供了坚实的理论基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Expression, DNA methylation pattern and transcription factor EPB41L3 in gastric cancer: a study of 262 cases.

Purpose: DNA methylation prominently inactivates tumor suppressor genes and facilitates oncogenesis. Previously, we delineated a chromosome 18 deletion encompassing the erythrocyte membrane protein band 4.1-like 3 (EPB41L3) gene, a progenitor for the tumor suppressor that is differentially expressed in adenocarcinoma of the lung-1 (DAL-1) in gastric cancer (GC).

Methods: Our current investigation aimed to elucidate EPB41L3 expression and methylation in GC, identify regulatory transcription factors, and identify affected downstream pathways. Immunohistochemistry demonstrated that DAL-1 expression is markedly reduced in GC tissues, with its downregulation serving as an independent prognostic marker.

Results: High-throughput bisulfite sequencing of 70 GC patient tissue pairs revealed that higher methylation of non-CpGs in the EPB41L3 promoter was correlated with more malignant tumor progression and higher-grade tissue classification. Such hypermethylation was shown to diminish DAL-1 expression, thus contributing to the malignancy of GC phenotypes. The DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-aza-CdR) was found to partially restore DAL-1 expression. Moreover, direct binding of the transcription factor CDC5L to the upstream region of the EPB41L3 promoter was identified via chromosome immunoprecipitation (ChIP)-qPCR and luciferase reporter assays. Immunohistochemistry confirmed the positive correlation between CDC5L and DAL-1 protein levels. Subsequent RNA-seq analysis revealed that DAL-1 significantly influences the extracellular matrix and space-related pathways. GC cell RNA-seq post-5-Aza-CdR treatment and single-cell RNA-seq data of GC tissues confirmed the upregulation of AREG and COL17A1, pivotal tumor suppressors, in response to EPB41L3 demethylation or overexpression in GC epithelial cells.

Conclusion: In conclusion, this study elucidates the association between non-CpG methylation of EPB41L3 and GC progression and identifies the key transcription factors and downstream molecules involved. These findings enhance our understanding of the role of EPB41L3 in gastric cancer and provide a solid theoretical foundation for future research and potential clinical applications.

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来源期刊
CiteScore
11.00
自引率
0.00%
发文量
180
期刊介绍: Cell Communication and Signaling (CCS) is a peer-reviewed, open-access scientific journal that focuses on cellular signaling pathways in both normal and pathological conditions. It publishes original research, reviews, and commentaries, welcoming studies that utilize molecular, morphological, biochemical, structural, and cell biology approaches. CCS also encourages interdisciplinary work and innovative models, including in silico, in vitro, and in vivo approaches, to facilitate investigations of cell signaling pathways, networks, and behavior. Starting from January 2019, CCS is proud to announce its affiliation with the International Cell Death Society. The journal now encourages submissions covering all aspects of cell death, including apoptotic and non-apoptotic mechanisms, cell death in model systems, autophagy, clearance of dying cells, and the immunological and pathological consequences of dying cells in the tissue microenvironment.
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