摘要
目的 探讨神经管缺陷（NTDs）家系中全基因组DNA甲基化改变情况及其意义。方法 收集3个NTDs家系，共12例研究对象纳入本研究，以NTDs患者作为病例组，家系中表型正常的成员为对照组。抽取12例研究对象的外周血，提取基因组DNA。使用重亚硫酸盐处理基因组DNA，全基因组扩增方式扩增片段化，置Illumina Infinium人类甲基化450k芯片上，进行杂交、洗脱、延伸、成像，使用iScan软件扫描芯片，Genome Studio读取扫描图像结果。使用Illumina甲基化分析器软件进行数据分析。结果 基因差异甲基化分析显示：差异性甲基化基因位点仅占检出CpG位点的0.2%，617个差异性DNA高甲基化的位点（P〈0.05），其中63个DNA甲基化位点P〈1×10－4，包括E盒结合锌指蛋白2基因、5，10－亚甲基四氢叶酸脱氢酶1基因等；104个差异性DNA低甲基化的位点（P〈0.05），其中65个DNA甲基化位点P〈0.01，包括同源异型盒B7基因、runt相关的转录因子3基因等。聚类分析显示：NTDs患儿呈DNA高甲基化改变的趋势较一致，而对照组呈DNA低甲基化改变的趋势较一致。结论 在NTDs家系中，DNA甲基化异常可能是NTDs发生的危险因素之一。
Objective To investigate the methylation alteration of genomic DNA （gDNA） and its significance in pedigree neural tube defects （NTDs）.Methods Twelve subjects from 3 NTDs pedigrees were enrolled in this study.NTDs patients were served as the case group, and their family members with normal phenotype were served as the control group.Peripheral vein blood was extracted, then gDNA was extracted.The extracted gDNA was treated with sodium bisulfite propagated as DNA segments in the way of whole genome amplification, which was put in Illumina Infinium human methylation 450k bead chip to perform hybridization, elution, extension, and imaging.The chip was scanned by iScan.Genome Studio was used to read the outcome.Illumina methylation analyzer software was used to analyze the methylation data.Results Gene differential methylation analysis showed that differential methylation sites only accounted for 0.2% of the detected CpG sites and there were 617 differential hypermethylation sites （P〈0.05）, and 63 of them represented significant difference（P〈1×10-4）, including zinc finger E-box binding homebox 2, 5, 10-methylenetetrahydrofolate dehydrogenase 1 etc; there were 104 differential hypomethylation sites（P〈0.05）, and 65 of them represented significant difference（P〈0.01）, including Homeobox B7 and runt-related transcription factor 3 etc.Clustering analysis indicated that the tendency of DNA hypermethylation was consistent with NTDs patients, but the tendency of DNA hypomethylation was consistent with the controls.Conclusion In NTDs pedigree, the abnormal DNA methylation alterations may be the risk factor for NTDs occurrence.
出处
《中华实用儿科临床杂志》
CSCD
北大核心
2017年第18期1420-1424,共5页
Chinese Journal of Applied Clinical Pediatrics
基金
国家自然科学基金（8177589）
国家重点基础研究发展计划（973计划）（2013CB945404）
天津市应用基础与前沿技术研究计划（14JCYBJC25000）
天津市卫生行业重点攻关项目（16KG166）
