Yu Song, Minghang Yu, Yuan Zhang, Xi Wang, Xiangyi Liu
{"title":"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","authors":"Yu Song, Minghang Yu, Yuan Zhang, Xi Wang, Xiangyi Liu","doi":"10.1002/eer3.70020","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Background</h3>\n \n <p>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 <i>IL8</i> regulation have been reported, the epigenetic landscape governing <i>IL8</i> expression in CRS remains unexplored.</p>\n </section>\n \n <section>\n \n <h3> Objective</h3>\n \n <p>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.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>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 <i>IL8</i> promoter region under TNF-α stimulation.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>TNF-α stimulation induced time and concentration-dependent upregulation of IL-8 mRNA (<i>p</i> < 0.001) and protein secretion (<i>p</i> < 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 (<i>p</i> < 0.001) and protein levels (<i>p</i> < 0.01). ChIP-qPCR confirmed TNF-α-dependent enrichment of H3K27me3 at the <i>IL8</i> promoter, supporting EZH2-mediated transcriptional repression.</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>EZH2-dependent H3K27 trimethylation is a key epigenetic mechanism controlling IL-8 gene expression in HNEpCs.</p>\n </section>\n </div>","PeriodicalId":100519,"journal":{"name":"Eye & ENT Research","volume":"2 3","pages":"194-201"},"PeriodicalIF":0.0000,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/eer3.70020","citationCount":"0","resultStr":"{\"title\":\"EZH2 negatively regulates IL-8 expression in human nasal epithelial cells through its histone methyltransferase activity\",\"authors\":\"Yu Song, Minghang Yu, Yuan Zhang, Xi Wang, Xiangyi Liu\",\"doi\":\"10.1002/eer3.70020\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Background</h3>\\n \\n <p>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 <i>IL8</i> regulation have been reported, the epigenetic landscape governing <i>IL8</i> expression in CRS remains unexplored.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Objective</h3>\\n \\n <p>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.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>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 <i>IL8</i> promoter region under TNF-α stimulation.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>TNF-α stimulation induced time and concentration-dependent upregulation of IL-8 mRNA (<i>p</i> < 0.001) and protein secretion (<i>p</i> < 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 (<i>p</i> < 0.001) and protein levels (<i>p</i> < 0.01). ChIP-qPCR confirmed TNF-α-dependent enrichment of H3K27me3 at the <i>IL8</i> promoter, supporting EZH2-mediated transcriptional repression.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusion</h3>\\n \\n <p>EZH2-dependent H3K27 trimethylation is a key epigenetic mechanism controlling IL-8 gene expression in HNEpCs.</p>\\n </section>\\n </div>\",\"PeriodicalId\":100519,\"journal\":{\"name\":\"Eye & ENT Research\",\"volume\":\"2 3\",\"pages\":\"194-201\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2025-07-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/eer3.70020\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Eye & ENT Research\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/eer3.70020\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Eye & ENT Research","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/eer3.70020","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
EZH2 negatively regulates IL-8 expression in human nasal epithelial cells through its histone methyltransferase activity
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.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
