Genes regulated by both of Ten-Eleven translocation 3 and Tumor necrosis factor α in Rheumatoid arthritis fibroblast-like synoviocytes. Genes regulated by both of Ten-Eleven translocation 3 and Tumor necrosis factor α in Rheumatoid arthritis fibroblast-like synoviocytes

As DNA demethylation is a dynamic epigenetic regulation in pattern formation of DNA methylation, we reasoned that DNA demethylation facilitates Rheumatoid arthritis (RA) progression. To address this point, we picked up the DNA dioxygenase family members (Ten-Eleven translocation: TET1/2/3) as these enzymes conduct the prime process of DNA demethylaion. Tumor necrosis factor α (TNFα) , one of the pro-inflammatory cytokines, was found effective in up-regulation in TET3 level in the cultured fibroblast-like synoviocytes (FLS), and the stimulated FLS cells exhibited high cell mobility with gene induction of cell-migration related factors in a TET3-dependent manner. From our findings, we presume that DNA demethylation mediated TET3 facilitates RA progression and chronicity as an epigenetic regulation. Overall design: To ask whether TET3 mediates the TNFα action in the gene expression program for the transformation of FLS, we used a gene microarray analysis to profile global gene regulation in the cultured RA FLS treated by TNFα, together with TET3 knockdown by small interfering RNA (siRNA).

DNA去甲基化（DNA demethylation）是DNA甲基化模式形成过程中的动态表观遗传调控机制，我们据此推测DNA去甲基化可促进类风湿关节炎（Rheumatoid arthritis, RA）的进展。为验证这一假说，我们选取了介导DNA去甲基化核心过程的DNA双加氧酶家族成员——十-十一易位酶（Ten-Eleven translocation, TET1/2/3）。研究发现，促炎细胞因子（pro-inflammatory cytokines）之一的肿瘤坏死因子α（Tumor necrosis factor α, TNFα）可上调培养的类风湿关节炎成纤维样滑膜细胞（fibroblast-like synoviocytes, FLS）中TET3的表达水平；经TNFα刺激的FLS细胞迁移能力显著增强，且该效应以TET3依赖的方式，通过诱导细胞迁移相关基因的表达实现。基于上述研究结果，我们推测TET3介导的DNA去甲基化作为一种表观遗传调控机制，可促进类风湿关节炎的进展与慢性化。整体实验设计：为探究TET3是否介导TNFα对成纤维样滑膜细胞表型转化相关基因表达程序的调控作用，我们采用基因芯片分析（gene microarray analysis）技术，对经TNFα处理、同时通过小干扰RNA（small interfering RNA, siRNA）敲低TET3的培养类风湿关节炎FLS细胞进行全局基因调控谱分析。