胚鼠腭裂形成过程中腭胚组织全基因组DNA甲基化的研究  

Genome-wide DNA methylation of palate embryonic tissue in process of embryonic mouse cleft palate formation

作　　者：刘丹 舒申友[1] 李珂[1] 舒璇 董泽君 郎兴 LIU Dan;SHU Shen-you;LI Ke;SHU Xuan;DONG Ze-jun;LANG Xing(Department of Burn and Plastic Surgery,The Second Affiliated Hospital of Shantou University Medical College,Shantou 515041,China)

机构地区：[1]汕头大学医学院第二附属医院烧伤整形外科,广东汕头515041

出　　处：《中国病理生理杂志》2018年第9期1638-1647,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81001284);广东省自然科学基金资助项目(No.2015A030313431)

摘　　要：目的:分析胚鼠腭裂形成过程中腭胚组织全基因组DNA差异甲基化位点相关基因及基因组差异甲基化水平。方法:使用高效低成本全基因组DNA甲基化检测技术——甲基化修饰依赖性内切酶测序法(MethylRAD-Seq法)对妊娠第13.5天(GD13.5)、GD14.5和GD16.5(n=18)的胚鼠腭裂模型组和对照组腭胚组织进行全基因组DNA甲基化测序,比较基因组不同功能元件(3’端非翻译区、5’端非翻译区、TSS2000、内含子、外显子和基因间区)中甲基化位点的差异分布及甲基化水平差异基因,并对相关基因进行GO和KEGG通路分析。结果:(1)DNA不同元件的甲基化位点的峰值个数和峰值,对照组无明显变化,实验组甲基化整体水平随着时间推移逐渐降低;(2)GO功能富集和KEGG通路富集分析差异甲基化位点的基因及甲基化水平有差异的相关基因,其中与腭裂形成相关的基因为Fgf16、Jarid2、Kdm6a、Nr3c1、Tbx22和Trp53;(3)与腭裂形成相关的信号通路主要有MAPK、Wnt和TGF-β信号通路。结论:DNA甲基化在腭裂形成过程中可能起重要的调控作用。AIM:To analyze genome-wide DNA differential methylation site-related genes and genomic diffe-rential methylation levels involved in embryonic mouse cleft palate formation.METHODS:MethylRAD-Seq was performed on the gestational day 13.5(GD13.5),GD14.5 and GD16.5 in experimental group and control group,which is a method for genome-wide DNA methylation profiling using methylation-dependent restriction enzymes.A comparison was made to process the different location of the methylation site and the methylation level differential genes among genome-wide different functional components(3’-untranslated region,5’-untranslated region,TSS2000,intron,exon,and intergenic region).The genome-wide DNA differential methylation site-related genes and methylation differential genes were analyzed using GO function enrichment and KEGG pathway enrichment.RESULTS:(1)The peak number and peak value of methylation sites in different components of DNA did not change significantly in the control group,and the methylation level of the experimental group decreased gradually over time.(2)GO function enrichment analysis and KEGG pathway enrichment ana-lysis were performed to process methylation level differential genes and genome-wide DNA differential methylation site-rela-ted genes,among which there were 6 genes(Fgf16,Jarid2,Kdm6a,Nr3c1,Tbx22 and Trp53)involved in embryonic mouse cleft palate formation.(3)MAPK,Wnt,TGF-βand other signaling pathways were found to participate in the formation of cleft palate.CONCLUSION:DNA methylation plays an important regulatory role in the process of cleft palate formation.

关 键 词：DNA甲基化 腭裂 MethylRAD-Seq 富集分析 

分 类 号：R329.1[医药卫生—人体解剖和组织胚胎学] R363.2[医药卫生—基础医学]