文章摘要
张慧文,周国平.曲谷抑霉素A调控人干扰素调节因子3基因初步研究[J].南京医科大学学报,2017,(1):20~24转120
曲谷抑霉素A调控人干扰素调节因子3基因初步研究
Trichostain A transcriptionally induces interferon regulatory factor 3 expression
投稿时间:2016-04-20  
DOI:10.7655/NYDXBNS20170104
中文关键词: 曲谷抑霉素A  人干扰素调节因子3  乙酰化  p300  基因转录调控
英文关键词: trichostain A  interferon regulatory factor 3  acetylation  p300  transcriptional regulation
基金项目:国家自然科学基金(81170661,81300023);南京市科技计划项目(201503003)
作者单位
张慧文 南京医科大学第一附属医院儿科江苏 南京 210029 
周国平 南京医科大学第一附属医院儿科江苏 南京 210029 
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中文摘要:
      目的:探讨组蛋白去乙酰化酶抑制剂(histone deacetylase inhibitor, HDACi)曲谷抑霉素A(trichostain A, TSA)对人干扰素调节因子3(interferon regulatory factor 3, IRF3)基因表达的影响,初步分析TSA调控IRF3基因的乙酰化机制。方法:双荧光素酶活性分析法检测TSA和组蛋白乙酰化酶(histone acetylase, HAT)野生型p300对IRF3基因启动子活性的影响;实时荧光定量PCR检测TSA对IRF-3基因mRNA水平的影响;Western blot检测TSA对IRF3蛋白水平的影响。结果:不同浓度TSA干预后IRF3基因启动子活性均增加,TSA浓度为1 μmol/L时,PS1、PS6相对荧光素酶活性分别增加75%、155%;过表达野生型p300后PS1相对荧光素酶活性增加267%;TSA(1、10 μmol/L)干预细胞6 h后,IRF3基因mRNA水平增加23%、39%,干预24 h后,mRNA水平增加17%、64%;TSA(0.1、1、5 μmol/L)干预细胞24 h后,蛋白表达量分别增加15%、72%、191%。结论:TSA上调IRF3基因mRNA和蛋白表达,TSA和组蛋白乙酰化酶p300均可正向调控IRF3基因启动子活性,TSA可能通过IRF3通路发挥治疗免疫性疾病和抗肿瘤作用。
英文摘要:
      Objective:To explore the influence of histone deacetylase inhibitor (HDACi) trichostain A (TSA) on the expression of interferon regulatory factor 3 (IRF3) and the preliminary epigenetic mechanism of transcriptional regulation. Methods:Luciferase assays were applied to detect IRF3 promoter activity after TSA incubation and histone acetyltransferases (HAT) p300 transfection. The IRF3 mRNA expression level was detected by Real-time fluorescence quantification-PCR, while the IRF3 protein expression level was detected by Western blot. Results: IRF3 gene promoter activity was increased after TSA intervention at different concentrations. TSA treatment (1 μmol/L) induced the relative luciferase activity (RLA) of IRF3 promoter PS1 and PS6 by 75% and 155%, respectively. Post-transfection with p300 increased the RLA of PS1 by 267%. At the 1 μmol/L and the 10 μmol/L TSA group, the IRF3 mRNA expression was increased by 23% and 39% after 6 h treatment, while increased by 7% and 64% after 24 h, respectively. The IRF3 protein expressions were increased by 15%, 72%, and 191% after TSA (0.1 μmol/L,1 μmol/L, and 5 μmol/L) treatment, respectively. Conclusion:TSA can up-regulate the expression of IRF3 both on mRNA and protein levels, while TSA and p300 can increase transcription activity by targeting IRF3 promoter region, which suggests that HDAC and acetylation are positive regulation of IRF3 transcription or expression, and indicates the hypothesized role of TSA in the immunotherapy and antitumor effects through IRF3 pathway.
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