噪声性听觉损伤易感豚鼠差异表达基因的筛选与分析

目的利用抑制消减杂交技术(SSH)筛选分析噪声性听觉损伤易感与不易感豚鼠耳蜗差异表达基因。方法易感组与不易感组豚鼠耳蜗组织利用Trizol法分别提取总RNA;SmartTM技术分别合成cDNA;SSH筛选差异表达基因,并进行测序和生物信息学分析。结果筛选噪声性听觉损伤(NIHL)易感与不易感豚鼠耳蜗的明显差异表达基因片段148条,包括易感组豚鼠耳蜗表达上调73条和下调75条。同源性大于80%且匹配碱基大于100 bp的相关序列共34条,其中上调7条,包括多聚泛素基因、β珠蛋白和α珠蛋白等;下调27条,包括线粒体基因组(mtDNA)、热休克蛋白(HSP)和细胞周期非特异性因子1等。结论SSH是一种高效的筛选差异表达基因的方法,mtDNA和HSP与NIHL及其易感性的发生相关。

Screening and analysis on differential expressed genes of NIHL susceptible guinea pigs

Objective To screen and analyze differentially expressed genes of cochlea between noise induced hearing loss(NIHL) susceptible and unsusceptible guinea pigs by using suppression subtractive hybridization(SSH).Methods Total mRNA of the susceptible group and the unsusceptible group were extracted by the Trizol method.cDNA of the two groups was synthesized by SmartTM technology and differential expressed genes were screened by SSH method.Finally,the positive clones were sequenced and a preliminary bioinformatics analysis was made.Results 148 differentially expressed genes of cochlea between NIHL susceptible and unsusceptible guinea pigs were screened with 73 up-regulation and 75 down-regulation.34 of them were relative sequences and their homology was larger than 80% and match sequence larger than 100 bp,7 of which were up-regulation,including UbB genes for polyubiquitin B,beta-globin mRNA and alpha-globin mRNA,etc.Concurrently,there were also 27 down regulation,including mitochondrial genome,heat shock protein mRNA and CCNB1 mRNA,etc.Conclusion SSH was a highly effective method for screening differential expressed genes,the mitochondrial genome and the heat shock protein were correlated with the occurrence of NIHL and its susceptibility.