体外培养耳蜗毛细胞氧化应激损伤的微小RNA调控网络分析  

Analysis of microRNA regulatory network in cochlear hair cells with oxidative stress injury

作　　者：王军义[1] 夏源[1] 杨翠婵[1] 王致[2] 

机构地区：[1]广东药科大学公共卫生学院劳动卫生与环境卫生学系,广州510310 [2]广州市第十二人民医院职业卫生检测中心,510620

出　　处：《中华耳鼻咽喉头颈外科杂志》2016年第10期751-755,共5页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：基金项目：广东省科技计划项目（2014A0202126）

摘　　要：目的 通过体外培养耳蜗毛细胞氧化应激损伤的差异性微小RNA（microRNA,miRNA）构建的基因调控网络,分析耳蜗毛细胞氧化应激损伤的重要基因及功能,探讨以氧化损伤为基础的多种感音性聋的分子机制.方法 以200 μmol/L叔丁基双氧气水（tert butyl hydrogen peroxide,t-BHP）染毒培养的小鼠HEI-OC1细胞系构建氧化应激损伤耳蜗毛细胞模型,高通量sRNA测序筛选差异性miRNA,以6个差异性表达最显著的miRNA进行调控网络构建,分析miRNA调控网络互作关系里面的重要基因,并通过Gene Cards数据库对重要基因进行功能注释.结果 高通量sRNA测序发现氧化应激损伤毛细胞出现24个差异性miRNA,其中mir-1934（logFC=2.367 947,P=2.35×10^-7）、mir-411（logFC=2.093 687,P=3.13×10^-6）、mir-717（logFC=1.927 67,P=3.24×10^-5）、mir-503 （logFC=-2.021 45,P =3.07×10^-6）、mir-467e（logFC=-1.953 28,P =0.000 137）、mir-699o（logFC=-1.950 06,P=0.000517）差异最显著;构建的miRNA调控网络中相连基因个数超过50节点的重要基因共有11个：Akt1、Src、Ctnnb1、Creb1、Ccnd1、Egfr、Gsk3b、Pten、Cdh1、Fras1、Ccnd2,其主要功能体现在参与多种细胞内信号传导通路调控细胞的凋亡和增殖.结论 氧化应激损伤毛细胞有多种信号传导通路（PI3 K-AKT/PKB通路、AKT/PKB通路、Wnt通路、ERK通路、Ras通路）参与调控细胞增殖和凋亡过程,这些信号通路互相交联形成网络调控,其中PI3K-AKT/PKB信号通路最为重要.Objective To analysis the important genes and functions of cochlear hair cells with oxidative stress injury,by the construction of gene regulatory network which based on different miRNA in cochlear hair cells in vitro with oxidative stress injury,and to explore the molecular mechanisms of deafness based on oxidative stress injury.Method The oxidative stress damage cochlear hair cell model was induced by 200 μmol/L t-BHP exposure in vitro.Small RNA deep sequencing analyzed the difference expression of miRNA and contructed gene regulatory network by 6 most significant difference miRNA.The important interaction genes in regulatory network were screened and important genes function were annotated by GeneCards.Result There were 24 different miRNAs in cochlear hair cells with oxidative stress injury by sRNA deep sequencing.Six most significant difference miRNA were：mir-1934 （logFC =2.367 947,P =2.35 × 10^-7）,mir-411 （logFC =2.093 687,P =3.13 × 10^-6）,mir-717 （logFC =1.927 67,P =3.24 × 10-5）,mir-503 （logFC =-2.021 45,P =3.07 × 10^-6）,mir-467e （logFC =-1.953 28,P =0.000 137）,and mir-699o （logFC =-1.950 06,P =0.000 517）.Eleven important genes in miRNA regulatory network were：Akt1,Src,Ctnnb1,Creb1,Ccnd1,Effr,Gsk3b,Pten,Cdh1,Fras1,and Ccnd2.Their main functions were to regulate hair cells apoptosis and proliferation by different intracellular signaling pathways.Conclusion There are many signaling pathways （PI3K-AKT/PKB signaling pathway,AKT/PKB signaling pathway,Wnt signaling pathway,ERK signaling pathway,and Ras signaling pathway） involved in the regulation of apoptosis and proliferation in cochlear hair cells with oxidative stress injury and these signaling pathways are linked to each other to form a network.PI3K-AKT/PKB signaling pathway seems to be the most active in cochlear hair cells with oxidative stress injury.

关 键 词：氧化性应激 毛细胞 听觉 高通量核苷酸测序 MicroRNA调控网络 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]