DNA甲基化与组蛋白去乙酰化对人类白细胞抗原-B27基因表达调控的研究  

The relationship between DNA methylation and histone deacetylation with HLA-B27 gene expression

作　　者：赵丽珂[1] 古洁若[1] 余得恩[2] 

机构地区：[1]中山大学附属第三医院风湿科,广州510630 [2]加州大学洛杉矶分校风湿科

出　　处：《中华风湿病学杂志》2008年第5期299-303,共5页Chinese Journal of Rheumatology

基　　金：国家自然科学基金杰出青年科学基金（30325019）

摘　　要：目的研究DNA甲基化酶抑制剂和组蛋白去乙酰化酶抑制剂对人类白细胞抗原（HLA）-B27基因表达的调控作用。方法构建表达Hela—B27启动子的Hela稳定转染细胞株，使用荧光素酶检测DNA甲基化酶抑制剂（5-Aza-CdR）和组蛋白去乙酰化酶抑制剂（HDACI）[丁酸钠（NaB），曲古霉素（TSA），丙戊酸（VPA）和Hydralazine]对HLA—B27启动子活性的影响，及其各种被抗炎症细胞因子抗体[抗肿瘤坏死因子（TNF）-α，抗干扰素（IFN）-β，抗IFN—γ，Infiximab]对其调节作用。然后使用5-Aza—CdR和HDACI培养表达HLA—B27基因组DNA的CCL6稳定细胞株，使用实时定量聚合酶链反应（PCR）技术检测HLA—B27mRNA表达水平。结果NaB和VPA可使HLA—B27启动子活性明显增高（24．0±1．7）倍和（17．4±2．2）倍（P〈0．05），且VPA和TNF—α，IFN-α，IFN—β，IFN—γ和佛波酯（PMA）对HLA—B27启动子活性有协同诱导作用。实时定量PCR结果显示NaB，TSA，VPA和5-Aza—CdR均可使HLA—B27mRNA水平明显增加。结论DNA甲基化与组蛋白去乙酰化可明显上调HLA—B27基因表达水平。Objective To study the relationship between DNA methylation inhibitor and histone deacetylase inhibitor （HDACI） with HLA-B27 gene expression. Methods Hela-HLA-B27 promoter stable cell line was developed as the first step and followed by detecting the effect of DNA methylation inhibitor （5-Aza-CdR） and HDACI. The synergetic effect of HDACI and cytokines as well as the effect of anti-TNF-α antibody, anti-IFN-β antibody, anti-IFN-γ antibody and Infiximab on HLA-B27 promoter activity were tested by measuring luciferase value. Then HLA-B27 mRNA expression level in CCL6-B27 genomic DNA stable cell line was measured by real-time PCR. Results Sodium butyrate （NAB） and valproic acid （VPA） could significantly up-regulate HLA-B27 promoter activity at 24 h （24.0± 1.7） times, （17.4±2.2） times respectively, （P〈0.05）. The synergetic effect between VPA with TNF-α, IFN-α, IFN-β, IFN-γ and PMA on HLA-B27 promoter activity was found. None of the antibody could inhibit the effect of HDACI. It also showed that NaB, TSA, VPA and 5-Aza-CdR could significantly increase HLA-B27 mRNA expression in CCL6-B27 stable cell line. Conclusion DNA methylation and HDACI can up-regulate HLA-B27 gene expression level.

关 键 词：甲基化 组蛋白去乙酰化酶 人类白细胞抗原-B27 基因表达 

分 类 号：R593.23[医药卫生—内科学] R733.7[医药卫生—临床医学]