EZH2 negatively regulates IL-8 expression in human nasal epithelial cells through its histone methyltransferase activity

Eye & ENT Research Pub Date : 2025-07-15 DOI:10.1002/eer3.70020

Yu Song, Minghang Yu, Yuan Zhang, Xi Wang, Xiangyi Liu

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Abstract

Background

Chronic rhinosinusitis (CRS), characterized by persistent inflammation of the nasal and sinus mucosa, exhibits an escalating global prevalence and incidence. Interleukin-8 (IL-8), a key chemokine driving neutrophil recruitment, is implicated in CRS pathogenesis. While non-epigenetic mechanisms of IL8 regulation have been reported, the epigenetic landscape governing IL8 expression in CRS remains unexplored.

Objective

This study aimed to investigate the epigenetic regulation of IL-8 expression in human nasal epithelial cells (HNEpCs) with a focus on histone modification-mediated mechanisms.

Methods

Tumor necrosis factor-alpha (TNF-α) was selected as a prototypical pro-inflammatory stimulus through systematic screening. An in vitro model of IL-8 induction was established and validated in TNF-α-treated HNEpCs. Regulatory mechanisms were probed using bioinformatics tools (UCSC Genome Browser, Cistrome DB) and pharmacological inhibitors targeting histone-modifying enzymes. siRNA-mediated enhancer of zeste homolog 2 (EZH2) knockdown to assess its regulatory role in IL-8 expression; Chromatin Immunoprecipitation followed by quantitative PCR (ChIP-qPCR) to determine whether H3K27me3 is directly enriched at the IL8 promoter region under TNF-α stimulation.

Results

TNF-α stimulation induced time and concentration-dependent upregulation of IL-8 mRNA (p < 0.001) and protein secretion (p < 0.001) in HNEpCs. TNF-α-mediated IL-8 upregulation was abrogated by the addition of methyltransferase inhibitors EPZ005687, EPZ6438, and BIX01294. SiRNA-mediated EZH2 depletion significantly enhanced both IL-8 mRNA (p < 0.001) and protein levels (p < 0.01). ChIP-qPCR confirmed TNF-α-dependent enrichment of H3K27me3 at the IL8 promoter, supporting EZH2-mediated transcriptional repression.

Conclusion

EZH2-dependent H3K27 trimethylation is a key epigenetic mechanism controlling IL-8 gene expression in HNEpCs.

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EZH2通过其组蛋白甲基转移酶活性负调控IL-8在人鼻上皮细胞中的表达

背景：慢性鼻窦炎（CRS）以鼻腔和鼻窦黏膜持续炎症为特征，在全球范围内呈现出不断上升的患病率和发病率。白细胞介素-8 （IL-8）是驱动中性粒细胞募集的关键趋化因子，参与CRS的发病机制。虽然已经报道了IL8调控的非表观遗传机制，但CRS中调控IL8表达的表观遗传格局仍未被探索。目的研究IL-8在人鼻上皮细胞（HNEpCs）中表达的表观遗传学调控，重点探讨组蛋白修饰介导的机制。方法通过系统筛选，选择肿瘤坏死因子-α (TNF-α)作为典型的促炎刺激因子。在TNF-α处理的HNEpCs中建立IL-8诱导的体外模型并进行验证。利用生物信息学工具（UCSC Genome Browser, Cistrome DB）和针对组蛋白修饰酶的药物抑制剂来探索调节机制。sirna介导的zeste同源物2增强子（EZH2）敲低对IL-8表达的调控作用染色质免疫沉淀后定量PCR （ChIP-qPCR）确定在TNF-α刺激下，H3K27me3是否在il - 8启动子区直接富集。结果TNF-α刺激诱导HNEpCs细胞IL-8 mRNA和蛋白分泌时间及浓度依赖性上调（p < 0.001）。添加甲基转移酶抑制剂EPZ005687、EPZ6438和BIX01294可消除TNF-α-介导的IL-8上调。sirna介导的EZH2缺失显著提高了IL-8 mRNA （p < 0.001）和蛋白水平（p < 0.01）。ChIP-qPCR证实了il - 8启动子上H3K27me3依赖TNF-α的富集，支持ezh2介导的转录抑制。结论ezh2依赖性H3K27三甲基化是控制IL-8基因表达的关键表观遗传机制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Eye & ENT Research

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