亚砷酸钠致人肝星状细胞纤维化及自噬中DNA甲基化位点筛查及差异甲基化谱构建  

Screening of DNA methylation sites and construction of differential methylation profiles in human hepatic stellate cells fibrosis and autophagy induced by sodium arsenite

作　　者：黄菲 迪丽娜尔·亚尔麦麦提 丁关鑫 赵丽君 周婧 吴顺华[1] Huang Fei;Dilinaer Yaermaimaiti;Ding Guanxin;Zhao Lijun;Zhou Jing;Wu Shunhua(School of Public Health,Xinjiang Medical University,Urumqi 830011,China;Department of Pathology,Xinjiang Medical University Cancer Hospital,Urumqi 830000,China)

机构地区：[1]新疆医科大学公共卫生学院,乌鲁木齐830011 [2]新疆医科大学附属肿瘤医院病理科,乌鲁木齐830000

出　　处：《中华地方病学杂志》2023年第1期11-16,共6页Chinese Journal of Endemiology

基　　金：国家自然科学基金(82160650);国家卫生健康委微量元素与地方病重点实验室开放课题;新疆维吾尔自治区研究生科研创新项目(XJ2022G165)。

摘　　要：目的分析亚砷酸钠(NaAsO_(2))致人肝星状细胞(LX-2细胞)纤维化及自噬相关的DNA甲基化位点,筛选与纤维化及自噬相关的特异性甲基化基因。方法采用DNA甲基化芯片Illumina Infinium Methylation EPIC BeadChips(850K甲基化芯片)对LX-2细胞(对照组)及NaAsO_(2)干预(低、中、高剂量组:终浓度分别为5、10、15μmol/L NaAsO_(2),干预48 h)致LX-2细胞纤维化及自噬模型进行全基因组DNA检测,寻找差异甲基化位点。对筛选出的差异甲基化基因进行GO功能富集分析和KEGG信号通路富集分析,了解基因功能。结果高剂量组细胞纤维化及自噬模型构建成功。850K甲基化芯片检测结果显示,高剂量组与对照组间存在25817个显著差异甲基化位点,其中高甲基化位点12083个、低甲基化位点13734个。GO功能富集分析显示,差异甲基化基因参与的分子功能主要包括蛋白结合、离子结合、催化活性、酶结合。KEGG信号通路富集分析显示,差异甲基化基因参与的通路主要包括代谢通路、癌症通路、磷脂酰肌醇-3-激酶-蛋白激酶B(PI3K-Akt)信号通路、内吞作用、丝裂原活化蛋白激酶(MAPK)信号通路。进一步在启动子区,筛选出11个与纤维化相关的差异甲基化基因和29个与自噬相关的差异甲基化基因。结论NaAsO_(2)诱导LX-2细胞纤维化及自噬过程中存在大量差异甲基化位点,筛选出与纤维化及自噬相关的特异性甲基化基因。Objective To analyze DNA methylation sites related to fibrosis and autophagy in human hepatic stellate cells(LX-2 cells)induced by sodium arsenite(NaAsO_(2)),and to screen specific methylation genes related to fibrosis and autophagy.Methods Genome-wide DNA detection was performed using Illumina Infinium Methylation EPIC BeadChips(850K methylation chip)to derive differential methylation sites in LX-2 cells(control group)and the fibrosis and autophagy models of LX-2 cells induced by NaAsO_(2)(low,medium and high dose groups:the final concentrations were 5,10,15μmol/L NaAsO_(2),respectively,after 48 h intervention).Gene ontology(GO)function enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)signaling pathway enrichment analysis were used to explore gene function.Results The model of cell fibrosis and autophagy was established successfully in high dose group.The results of 850K methylation chip detection showed that there were 25817 significant different methylation sites between the high dose group and the control group,including 12083 hypermethylation sites and 13734 hypomethylation sites.GO function enrichment analysis showed that the molecular functions of differentially methylated genes mainly included protein binding,ion binding,catalytic activity,enzyme binding.KEGG signaling pathway enrichment analysis showed that the pathways involved in differentially methylated genes mainly included metabolic pathway,cancer pathway,phosphatidylinositol-3-kinase-protein kinase B(PI3K-Akt)signaling pathway,endocytosis,and mitogen activated protein kinase(MAPK)signaling pathway.In the promoter region,11 and 29 differentially methylated genes related to fibrosis and autophagy were screened,respectively.Conclusions A large number of differential methylation sites exist in the process of NaAsO_(2) induced fibrosis and autophagy of LX-2 cells.Specific methylation genes related to fibrosis and autophagy are screened out.

关 键 词：亚砷酸盐类 人肝星状细胞 肝纤维化 自噬 DNA甲基化 

分 类 号：R575[医药卫生—消化系统]