基于生物信息学筛选阿片类药物依赖的关键基因的研究  

Bioinformatics analysis and validation of genes related to opioid addiction

作　　者：柯建林 向明亮 张伟[1] 邓小冬[1] 刘环[2] 章丽霞[1] 刘云[1,3] KE Jian-in;XIANG Ming-iang;ZHANG Wei;DENG Xiao-dong;LIU Huan;ZHANG Li-xia;LIU Yun(Institute of Basic Medicine and Forensic Medicine,North Sichuan Medical College,Nanchong 637000,Sichuan,China;School of Public Health,North Sichuan Medical College,Nanchong 637000,Sichuan,China;Medical Imaging Key Laboratory of Sichuan Province,North Sichuan Medical College,Nanchong 637000,Sichuan,China)

机构地区：[1]川北医学院基础医学与法医学院,四川南充637000 [2]川北医学院公共卫生学院,四川南充637000 [3]川北医学院医学影像四川省重点实验室,四川南充637000

出　　处：《中国药物依赖性杂志》2023年第3期206-212,共7页Chinese Journal of Drug Dependence

基　　金：四川省基层卫生发展研究中心项目(SWFZ19-Q-05);南充市科技局市校合作项目(20SXQT0212,20SXQT0327)。

摘　　要：目的基于生物信息学方法筛选并验证阿片类药物依赖的关键基因。方法Gene Expression Omnibus(GEO)数据库下载阿片类药物处理小鼠基因芯片数据集,采用GEO2R筛选差异表达基因(differentially expressed genes,DEGs),DAVID数据库进行GO和KEGG富集分析,蛋白-蛋白相互作用(protein-protein interaction,PPI)分析DEGs潜在生物学功能,MCODE和CytoHubba插件进行PPI可视化、筛选hub基因;实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)和蛋白免疫印迹(Western blotting,WB)检测海洛因处理BV2细胞株中hub基因及信号通路关键因子mRNA及蛋白表达水平。结果筛选出366个DEGs(上/下调157/209)。KEGG显示DEGs主要富集在PI3K/Akt信号通路。Spp1、Trp53、Egfr、Cd44是阿片类药物依赖的hub基因,涉及神经系统发育、细胞增殖、分化及凋亡等生物学过程。Spp1、Trp53、Cd44、PI3K及Akt mRNA和蛋白在海洛因处理BV2细胞株中表达增加,而Egfr mRNA和蛋白表达降低(P<0.05)。结论Spp1、Trp53、Egfr、Cd44、PI3K/Akt信号通路可能是阿片类药物依赖的hub基因及主要信号通路。Objective Identification and validation of key genes for opioid dependence based on bioinformatics approach.Methods Gene datasets from the Gene Expression Omnibus(CEO)database of opioid intervention mouse models were downloaded and screened for differentially expressed genes(DEGs)with CEO2R software.CO and KECG analysis and protein-protein interaction(PPI)were used to detect potential functions of DEGs,and Cytoscape plug-ins MCODE and CytoHubba to visualize all DEGs and filter for hub genes.The relative mRNA and protein expression of hub genes and key signaling pathway factors in heroin-treated BV2 cell lines were detected by Real-time quantitative PCR(RT-qPCR)and western blotting(WB),respectively.Results A total of 366 DEGs(up/down 157/209)were screened.KEGG analysis showed that DEGs was mainly enriched in PI3K/Akt signaling pathway.Sppl,Trp53,Egfr and Cd44 were hub genes of opioid dependence,which were mainly related to nervous system development,cell proliferation,differentiation,and apoptosis.The mRNA and protein of Sppl,Trp53,Cd44,PI3K,Akt were increased in BV2 cell treatment with heroin(P<0.05),while the mRNA and protein of Egfr was decreased(P<0.05).Conclusion Sppl,Trp53,Egfr,and Cd44,and PI3K/Akt signaling pathway were the hub genes and key signaling pathway of opioids dependence.

关 键 词：阿片类药物 依赖 生物信息学 GEO数据库 差异表达基因 

分 类 号：R749[医药卫生—神经病学与精神病学]