应用hMeDIP-chip技术分析系统性红斑狼疮患者5-羟甲基胞嘧啶状态  

Genome-wide analysis of DNA 5-hydroxymethylcytosine in uremia patients using hMeDIP-chip

作　　者：眭维国[1] 谭秋培[2] 林华[1] 刘行超[2] 陈洁晶[1] 薛雯[1] 戴勇[1] 

机构地区：[1]中国人民解放军第181医院中心实验室,广西代谢性疾病研究重点实验室,桂林541002 [2]中国人民解放军第181医院检验科,桂林541002

出　　处：《中华风湿病学杂志》2015年第9期618-622,共5页Chinese Journal of Rheumatology

基　　金：广西重点实验室建设项目，桂林市科技计划科技创新能力与条件建设

摘　　要：目的 研究SLE患者全基因组羟基化水平,寻找SLE患者与健康对照5-羟甲基胞嘧啶（5-hmC）水平差异显著的目的基因,作为SLE特异性生物标志物.方法 应用hMeDIP-chip技术,分别检测SLE患者15例和健康对照组15名外周血的DNA 5-hmC状态.通过GO功能分析、pathway分析等分析方法对比分析,筛选出羟甲基化水平差异显著的目的基因,并用实时定量PCR技术验证hMeDIP-chip结果可靠性.结果 NanoDrop ND-1000和琼脂糖凝胶电泳证明所提的样本总DNA质量良好,可用于后续实验.与健康对照组相比,SLE患者在启动子区有1 701个5-hmC差异基因,其中884个上调,817个下调;在CPG岛有3 826个5-hmC差异基因,其中2 034个上调,1 792个下调.挑选3个差异最大的基因：TREX1、CDKN1A、CDKN1B进行讨论,并进行实时定量PCR验证实验,其结果与hMeDIP-chip结果相符,证明了hMeDIP-chip结果的可靠性.结论 研究结果表明SLE患者的5-hmC状态有很大的变化,这些变化或许可以进一步揭示SLE的发病机制.这些新发现也表明了5-hmC可以作为SLE的潜在生物标志物和治疗靶点.Objective To investigate the role of the 5-hydroxymethylcytosine （5-hmC） DNA modification in the onset of systemic lupus erythemosus （SLE）,we compared tihe levels 5-hmC between SLE patients and normal controls.Methods With informed consent,whole blood was obtained from patients,and genomic DNA was extracted.Using hMeDIP-seq analysis and validation by quantitative real-time quantitative polymerase chain reaction （RT-PCR）,we identified the differentially hydroxymethylated regions that were associated with SLE.Results There were 1 701 genes with significantly different 5-hmC levels at the promoter region in the SLE patients compared with the normal controls.The CpG islands of 3 826 genes showed significant difference at 5-hmC levels in SLE patients compared with the normal controls.Out of the differentially hydroxymethylated genes,three were selected for validation,including TREX1,CDKN1A,and CDKN1B.The hydroxymethylation levels of these three genes were confirmed by quantitative RT-PCR.Conclusion Our studies indicate that there are significant alterations of 5-hmC in SLE patients;these differentially hydroxymethylated genes may contribute to the pathogenesis of SLE.Such novel findings show the significance of 5-hmC as a potential biomarker or promising target for epigenetic-based SLE therapies.

关 键 词：红斑狼疮 系统性 hMeDIP-chip 5-羟甲基胞嘧啶 

分 类 号：R593.241[医药卫生—内科学]