【作者】 黄晓蒂； 陈园； 田莎； 周青； 王志琪； 裴刚； 田雪飞；

【Author】 HUANG Xiao-di;CHEN Yuan;TIAN Sha;ZHOU Qin;WANG Zhi-qi;PEI Gang;TIAN Xue-fei;Medical experimental teaching centre of the traditional Chinese medicine college in Hunan University of Chinese Medicine;The Key Laboratory of Cell biology and molecular technology of the Combine traditional Chinese and western medicine institute in Hunan University of Chinese Medicine;The first hospital in Hunan University of Chinese Medicine;College of Pharmacy in Hunan University of Chinese Medicine;

【机构】 湖南中医药大学中医学院医学基础教学实验中心； 湖南中医药大学中西医结合学院细胞生物学与分子技术湖南省高校重点实验室； 湖南中医药大学第一附属医院； 湖南中医药大学药学院；

【摘要】 目的利用全基因组DNA甲基化芯片,检测蜈蚣提取物（Centipede Scolopendra extraction,CSE）处理后人肝癌细胞全基因组水平DNA甲基化谱情况,并对全基因表达谱芯片的差异基因进行注释系统（gene onto1ogy,G0）分析。方法 MTS法检测CSE对人肝癌细胞HCCLM3增殖的影响,并计算细胞增殖半数抑制率(IC₅₀)。以1/2 IC₅₀浓度的CSE连续处理72 h,设置空白对照组。收集细胞提取基因组DNA,采用全基因组甲基化芯片进行基因组DNA甲基化检测,分析筛选甲基化差异性位点。结果（1）MTS检测结果显示,CSE以剂量依赖方式抑制人肝癌细胞的生长,IC₅₀值为0.613 mg/mL。（2）全基因组甲基化芯片检测结果显示:①经CSE处理72h时,CSE处理组与空白组甲基化谱发生了明显变化;初筛差异明显有意义的甲基化位点有:EPHB4、UTRN、DHX15、H19基因;②经5-Azacytudine处理72 h时,5-Azacytudine组与CSE处理组比较甲基化谱差异,经过初筛,变化较为明显的位点有,FHL1、EIF4G1、DLX2、H19、EBF3基因。（3）对全基因表达谱芯片的差异基因进行注释系统（gene ontology,GO）分析,蜈蚣组与空白组比较总共有3847个基因表达水平发生了变化,其中上调1837个基因,下调2010个基因,G0分析结果发现这些改变发生在细胞线粒体、细胞膜以及胞浆中,参与代谢和转运等生物学过程。在Pathway分析结果中显示共有3个信号通路发生了显著性变化,具有显著性意义。更多还原

【Abstract】 Objective:Discuss the different of the genome-wide methylation research of malignant liver cancer cells which treated with the Centipede Scolopendra extraction（CSE） with the whole genome DNA methylation chip and analyse the differences of gene expression profile chip with gene annotation system（gene ontology,GO）.Methods:The cellular proliferation was assessed with MTS in liver cancer cells HCCLM3 used with the CSE.Calculate the IC50 alue concentration.CSE sequential processing with 1/2 IC₅₀ con-centration of the cells in 3 d,blank comparison group,Genome-wide methylation chip are used to observe drugs on human hepatocellular carcinoma HCCLM3 cells genome-wide methylation spectrum analyze data and the degree of Methylation differences sites for screening.Result:（1）CSE effect the HCCLM3,MTS results show that the extracts are inhibition of cell growth,Calculation drug intervention HCCLM3 IC50 value concentration of 0.613mg/mL.（2）Methylation chip according to the results:① Genome-wide chip shows blank group and CSE group have changed,the site located at EPHB4、 UTRN、DHX15、H19;②The 5-Azacytudine group and CSE group have obvious differences.the site located at FHL1,EIF4G1、 DLX2,H19,EBF3;（3）Methylation chip according to the results:analyse the differences of gene expression profile chip with gene annotation system（gene ontology,GO）.The centipede group compared with the blank group has3847 changed the level of gene expression,which raised 1837 genes,cut 2010 genes,GO analysis found that the change in cell mitochondria and cell membrane and cytoplasm,engaging in biological processes such as metabolism and transport.In the Pathway analysis results show that there are 3 significant changes have taken place in the signal path,has the significant significance.更多还原