摘要
目的探讨急性低氧诱导人肝癌细胞(Huh-7、Hep3B)差异表达基因的筛选及其功能。方法常规培养Huh-7、Hep3B细胞,随机取部分细胞置于常氧条件(空气、5%CO 2)下培养,取部分细胞置于低氧条件(1%O 2、5%CO 2、94%N 2)下培养,均培养20 h后收样。采用Gene Chip Clariom S Array芯片筛选急性低氧诱导和常氧处理细胞的差异表达基因,筛选条件为错误发现率<0.05且|log 2FC|>1;用Kolmogorov-Smirnov和KOBAS version 2.0对差异表达基因进行基因本体论功能分析和京都基因与基因组百科全书通路分析。结果与常氧处理的细胞相比,急性低氧诱导的Huh-7细胞中差异表达基因共计392个,其中表达上调基因266个,表达下调基因126个;急性低氧诱导的Hep3B细胞中,差异表达基因共计2260个,其中表达上调基因847个,表达下调基因1413个。急性低氧诱导的Huh-7、Hep3B细胞共同的差异表达基因共177个,表达上调基因127个,表达下调基因50个。急性低氧诱导的Huh-7细胞中差异表达基因主要与小分子代谢、有机物生物合成、细胞应激反应相关,主要富集在糖酵解糖异生、MAPK、细胞周期、癌症相关信号通路上。急性低氧诱导的Hep3B细胞中差异表达基因主要与小分子代谢、磷酸化代谢、细胞生长与凋亡等相关,主要富集在细胞周期、神经退行性疾病、细胞凋亡以及磷酸化调控相关信号通路上。两株细胞系共同的差异表达基因表达上调最明显的3个基因TXNIP、ATF3、SLC6A8均与肝癌发生相关。结论成功从急性低氧诱导人肝癌细胞中筛选出差异表达基因,呈高表达的基因可能与肝癌发生相关。
Objective To investigate the screening and function of differentially expressed genes in human hepatocellular carcinoma cell line(Huh-7 and Hep-3B)induced by acute hypoxia.Method Huh-7 and Hep3B cells were routinely cultured.Some cells randomly selected were cultured under normoxic conditions(air,5%CO2),and some cells were cultured under hypoxia conditions(1%O2,5%CO2,94%N2).After 20 hours of culture,the samples were collected.The differentially expressed genes in cells induced by acute hypoxia and normoxic treatment were screened out by Gene Chip Clariom S Array microarray under the conditions of error detection rate<0.005 and|log2FC|>1,and the gene ontology function analysis and Kyoto gene and genome encyclopedia pathway analysis were conducted by Kolmogorov-Smirnov and KOBAS version 2.0.Results Compared with the cells treated with normoxia,there were 392 differentially expressed genes in Huh-7 cells induced by acute hypoxia,of which,266 up-regulated genes and 126 down-regulated genes were expressed;2260 differentially expressed genes were expressed in Hep-3B cells induced by acute hypoxia,of which,847 were up-regulated and 1413 were down-regulated.There were 177 differentially expressed genes in both Huh-7 and Hep-3B cells induced by acute hypoxia,of which,127 were up-regulated and 50 were down-regulated.The differentially expressed genes in Huh-7 cells induced by acute hypoxia were mainly related to small molecular metabolism,organic biosynthesis and cell stress response,which were mainly enriched in glycolysis glycolysis,MAPK,cell cycle and cancer related signaling pathway.The differentially expressed genes in Hep-3B cells induced by acute hypoxia were mainly related to small molecule metabolism,phosphorylation metabolism,cell growth and apoptosis,and they were mainly enriched in cell cycle,neurodegenerative diseases,apoptosis and phosphorylation regulation signaling pathways.The top up-regulated three differentially expressed genes in the two cell lines were TXNIP,ATF3 and SLC6A8,which were all associated wi
作者
路远
韦花媚
谭川
吴贤建
邵泽生
伍毅
许佐明
李文川
浦涧
LU Yuan;WEI Huamei;TAN Chuan;WU Xianjian;SHAO Zesheng;WU Yi;XU Zuoming;LI Wenchuan;PU Jian(Youjiang Medical University for Nationalites,Baise 533000,China)
出处
《山东医药》
CAS
2020年第1期1-4,共4页
Shandong Medical Journal
基金
广西肝胆疾病临床医学研究中心研究课题(桂科AD17129025)
关键词
肝癌
低氧
人肝癌细胞系
基因芯片技术
差异表达基因
liver carcinoma
hypoxia
human hepatocellular carcinoma cell line
gene chip technique
differentially expressed gene
