摘要
目的分析不明原因的复发性流产(URSA)患者全基因组DNA甲基化差异,以探讨其新的病理机制。方法收集2021年1~10月于宁夏医科大学总医院就诊并明确诊断为URSA的3例患者作为URSA组,选取同期3例孕检无不良孕产史患者作为对照组,采用甲基化芯片技术,检测URSA患者外周血基因组DNA甲基化情况,初步筛查URSA患者的差异甲基化位点(DMCs),并对DMCs对应的基因使用GO(http://www.geneontology.org/)和KEGG(https://www.kegg.jp/kegg/pathway.html)进行生物信息学分析。结果URSA患者全基因组共筛选出39075个DMCs,其中33634个为高甲基化,5441个为低甲基化;启动子区域(Tss1500、Tss200、3′UTR、5′UTR、1stExon)有10941个(28.0%)DMCs,9797个为高甲基化,1144个为低甲基化。两组样本DMCs的层次聚类分析结果显示,URSA患者存在较多高甲基化位点,两组间存在显著的甲基化差异(P<0.05)。Go功能富集分析结果显示,生物进程主要富集在中性粒细胞活化及介导的免疫、细胞黏附正向调节;细胞组分主要富集在细胞连接、特异颗粒等;分子功能主要富集在丝氨酸/苏氨酸蛋白激酶活性、GTP酶活性调节、肌动蛋白结合等。KEGG分析结果显示,DMGs在多条信号通路上富集,其中主要富集在T细胞受体信号通路。结论URSA发生可能与DNA甲基化位点的改变有关,主要涉及到细胞免疫、黏附、酶活性调节等多个生物学过程和T细胞受体信号通路的调控。
Objective:To analyze whole genome DNA methylation difference in the patients with unexplained recurrent spontaneous abortion(URSA),and to explore the underlying mechanism.Methods:Three patients who were diagnosed with URSA at the General Hospital of Ningxia Medical University from January to October 2021 were selected as the URSA group.Three patients with no adverse pregnancy or childbirth history during the same period were recruited as the control group.The methylation chip technology was used to detect the genome DNA methylation of the peripheral blood of URSA patients.The differential methylation sites(DMCs)in URSA patients were screened preliminarily,and GO(http://www.geneontology.org/)and KEGG(https://www.kegg.jp/kegg/pathway.html)were used to conduct the bioinformatics analysis for the genes corresponding to DMCs.Results:A total of 39075 differential methylation sites were screened out in the URSA group,of which 33634 were hypermethylation and 5441 were hypomethylation.There were 10941(28.0%)DMCs in the promoter regions(Tss1500,Tss200,3′UTR,5′UTR,1stExon),9797 were hypermethylation and 1144 were hypomethylation.Hierarchical clustering analysis of DMCs in the two groups showed that there were more hypermethylation sites in URSA patients,and there was significant difference in methylation between the two groups(P<0.05).The results of Go function enrichment analysis showed that the biological process was mainly enriched in neutrophil activation,and neutrophil mediated immunity and positive regulation of cell adhesion.Cell components were mainly enriched in cell junction,specific particles,etc.Molecular functions were mainly enriched in serine/threonine protein kinase activity,GTPase activity regulation,actin binding,etc.The KEGG analysis results showed that DMGs were enriched in multiple signaling pathways,mainly in T cell receptor signaling pathways.Conclusions:The occurrence of URSA may be related to the changes of DNA methylation sites,mainly involving multiple biological processes such as cell immunity,adhesion,enzyme activity regulation and the regulation of T cell receptor signaling pathway.
作者
宋旭梅
詹福寿
范美荣
闫昕
王青
随瑞枝
SONG Xu-mei;ZHAN Fu-shou;FAN Mei-rong;YAN Xin;WANG Qing;SUI Rui-zhi(Clinical Laboratory Center,General Hospital of Ningxia Medical University,Yinchuan 750000)
出处
《生殖医学杂志》
CAS
2023年第6期904-911,共8页
Journal of Reproductive Medicine
基金
宁夏回族自治区自然科学基金(2019AAC03191)
宁夏回族自治区重点研发计划项目(2019BEG03054)。
关键词
复发性流产
DNA甲基化
生物信息学
Recurrent spontaneous abortion
DNA methylation
Bioinformatics analysis
作者简介
通讯作者:宋旭梅,女,宁夏银川人,硕士学历,主管技师,细胞与分子遗传专业。
