诱导山羊成纤维细胞转分化为乳腺上皮细胞的全基因组DNA甲基化分析  

Genomic DNA methylation analysis of goat fibroblasts induced to transdifferentiate into mammary epithelial cells

作　　者：孙哲 王国栋 刘权辉 雷志刚 黄奔 SUN Zhe;WANG Guodong;LIU Quanhui;LEI Zhigang;HUANG Ben(School of Animal Science and Technology,Guangxi University,Nanning 530004,China;Guangxi Zhuang Autonomous Region People's Hospital,Guangxi Medical Sciences,Nanning 530021,China)

机构地区：[1]广西大学动物科学技术学院,南宁530004 [2]广西医学科学院广西壮族自治区人民医院,南宁530021

出　　处：《黑龙江畜牧兽医》2024年第23期19-23,31,121,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31960160,31660342)。

摘　　要：为了获得山羊成纤维细胞转分化为乳腺上皮细胞过程的DNA甲基化水平和模式等表观遗传差异,试验采用全基因组甲基化测序(WGBS)技术对山羊成纤维细胞和转分化得到的乳腺上皮细胞进行DNA甲基化水平的检测和差异甲基化区域(DMR)分析,通过检测分析差异甲基化区域和差异甲基化基因解析甲基化参与山羊耳缘成纤维细胞转分化为乳腺上皮细胞的过程,探讨甲基化对乳腺上皮细胞发育调控的影响。结果表明:通过对WGBS文库数据的统计,共有2个测序样本的Q30值大于90.86%,分别被命名为DGFF5i8d和DGFF,即为诱导8 d转分化后的乳腺上皮细胞和山羊成纤维细胞。2个测序样本共检测到361个差异甲基化区域(其中130个上调差异甲基化区域和231个下调差异甲基化区域)和866个差异甲基化基因,对差异甲基化基因进行GO功能注释和KEGG信号通路富集分析,筛选出旁粒相关基因(PSPC1)、斑点型POZ蛋白(SPOP)、丝氨酸蛋白酶14(ST14)、X染色体连锁凋亡抑制因子相关因子(XAF1)、跨膜受体蛋白(Notch2)、黏着斑蛋白(VCL)、酸敏感离子通道(ASIC2)等7个与乳腺上皮发育相关的基因,并绘制出了成纤维细胞和乳腺上皮细胞全基因组甲基化图谱。说明本试验检测到的差异甲基化基因可能通过表观遗传调控机制参与调节乳腺发育过程。To investigate the epigenetic differences in DNA methylation levels and patterns in the process of transdifferentiation from goat fibroblasts to goat mammary cells,the whole genome methylation(WGBS)technology was used to assess the DNA methylation levels and differentially methylated regions(DMRs)between goat fibroblasts and mammary epithelial cells.By detecting and analyzing theDMRs and differentilly methylated genes(DMGs),how methylation was involved in the transdfferentiation from goat ear-derived fibroblasts to mammary epithelial cells was analyzed and its role in the regulation of mammary cell development was explored.The results showed that the Q30 values of the two samples were greater than 90.86%by the statistics of the WCBS library data,and the two samples were named DCFF5i8d and DGFF,namely mammary epithelial cells and goat fibroblasts after induced 8 d transdifferentiation,respectively.A total of 361 DMRs(130 up-regulated DMRS and 231 down-regulated DMRS)and 866 DMGs were identified by comparing the methylation levels of fibroblasts and mammary epithelial cells.Gene Ontology(GO)annotation and KECG pathway enrichment analysis were performed on the DMGs,and seven genes related to mammary epithelial development were identified:Paraspeckle Component 1(PSPC1),Speckle-type POZ Protein(SPOP),Serine Protease 14(ST14),XIAP-associated factor 1(XAF1),Transmembrane Receptor Protein(Notch2),adhesionplaque protein(VCL),and Acid-Sensing Ion Channel 2(ASIC2),and genome-wide methylation map of fibroblasts and mammary epithelial cells was constructed.It suggested that the DMGs identified in this study might participate in the regulation of mammary development by epigenetic regulatory mechanisms.

关 键 词：山羊 转分化 乳腺上皮细胞 全基因组 甲基化 

分 类 号：S826[农业科学—畜牧学] Q78[农业科学—畜牧兽医]