DENV2感染外周血单个核细胞增加TNF-α基因启动子区域CpG位点的甲基化水平  

Changes of DNA methylation at CpG sites in promoter region of TNF-α gene in DENV2-infected peripheral blood mononuclear cells

作　　者：张英可[1] 张林[1] 齐一鸣[1] 黄俊琪[1] 

机构地区：[1]中山大学中山医学院免疫学研究所,教育部热带病防治研究重点实验室,广东广州510080

出　　处：《中国病理生理杂志》2014年第1期150-153,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30872350;No.31370870);广东省自然科学基金资助项目(No.S201302001300;No.S2012010009050);广东省科技计划(No.2010B050700008;No.2011B040300022);广州市科技计划(No.2011J4100084);2010年中山大学高校基本科研业务费青年培育项目(No.10ykpy31)

摘　　要：目的:检测正常人外周血单个核细胞(PBMC)感染2型登革病毒(DENV2)后肿瘤坏死因子α(TNF-α)基因启动子区域CpG位点的甲基化水平。方法:采用亚硫酸氢盐测序PCR法检测DNA甲基化水平。结果:TNF-α基因启动子区域为-294 bp到+58 bp,覆盖11个散在CpG位点;PCR反应后取PCR产物进行琼脂糖凝胶电泳分析显示,扩增序列大小与理论预测相符合;PBMC感染DENV2 0 h和6 h在11个甲基化位点中有2个处于甲基化状态,感染12 h有6个甲基化位点甲基化。0 h、6 h和12 h的平均甲基化率分别为10.3%、12.1%和25.5%,且0 h和12 h及6 h和12 h的甲基化率差异有统计学意义。结论:PBMC感染DENV2后会引起TNF-α基因启动子区域的甲基化水平增加。AIM： To examine DNA methylation at CpG sites in the promoter region of tumor necrosis factor-al- pha （TNF-a） gene in dengue virus type 2 （DENV2） -infected peripheral blood mononuclear cells （PBMC）. METHODS： DNA methylation in the promoter region of TNF-a gene was measured by bisulfite sequencing PCR. RESULTS： The pro- moter region of TNF-a gene was from -294 bp to ＋ 58 bp, including 11 CpG sites. The PCR products identified by aga- rose gel electrophoresis were consistent with the theoretical size. Two sites were methylated at 0 h and 6 h and 6 sites were methylated at 12 h in TNF-a gene promoter region in DENV2-infected PBMC. The average methylation rates were 10. 3%, 12. 1% and 25.5% at 0 h, 6 h and 12 h, respectively. Significant differences between 0 h and 12 h and between 6 h and 12 h were observed. CONCLUSION： The DNA methylation in the promoter region of TNF-a gene is increased in DENV2- infected PBMCs.

关 键 词：登革病毒 肿瘤坏死因子仅 DNA甲基化 

分 类 号：R373.33[医药卫生—病原生物学]