曲谷抑霉素A调控人干扰素调节因子3基因初步研究

发布时间：2018-07-18 09:21

【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m RNA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1μmol/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、10μmol/L)干预细胞6 h后,IRF3基因m RNA水平增加23%、39%,干预24h后,m RNA水平增加17%、64%;TSA(0.1、1、5μmol/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m RNA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。
[Abstract]:Aim: to investigate the effect of histone deacetylase inhibitor (HDACi) on the expression of human interferon regulatory factor 3 (IRF3) gene, and to explore the mechanism of acetylation of human interferon regulatory factor 3 (interferon regulatory factor 3). Methods: double luciferase activity assay was used to detect the effects of TSA and histone acetylase (hat) wild type p300 on the activity of IRF3 gene promoter. The effect of TSA on the mRNA level of IRF-3 gene was detected by real-time fluorescence quantitative PCR and the effect of TSA on the level of IRF3 protein was detected by Western blot. Results: the promoter activity of IRF3 gene increased at 1 渭 mol / L TSA concentration of 1 渭 mol / L, and the relative luciferase activity of PS1 / PS6 increased 75 / 155, respectively, and the activity of PS1 relative luciferase was increased by 267% TSA (110 渭 mol / L) for 6 h after overexpression of wild type p300. After 24 hours of intervention, the level of mRNAs increased by 17% and 64% TSA (0.1 渭 mol / L) for 24 h. The protein expression increased by 15% and 72%, respectively. Conclusion TSA can up-regulate the mRNA and protein expression of IRF3 gene TSA and histone acetylase p300 can positively regulate the activity of IRF3 gene promoter TSA may play a role in the treatment of immune diseases and antitumor through IRF3 pathway.
【作者单位】：南京医科大学第一附属医院儿科;
【基金】：国家自然科学基金(81170661,81300023) 南京市科技计划项目(201503003)
【分类号】：Q78

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