肺癌消减杂交文库中共同基因的生物信息学分析及验证  

Bioinformatics Analysis and Verification of Common Genes in the Lung Cancer c DNA-libraries Constructed by Suppression Subtractive Hybridization

作　　者：吴明松[1] 樊萌萌[2] 马占山[2] 曹毅[3] 

机构地区：[1]遵义医学院细胞生物学与遗传学教研室,遵义563000 [2]中国科学院昆明动物研究所遗传资源与进化国家重点实验室,昆明650223 [3]中国科学院昆明动物研究所分子与实验病理学实验室,昆明650223

出　　处：《中国生物化学与分子生物学报》2015年第2期199-206,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展计划(973计划;2011CB510104);贵州省科技厅资助项目(黔科合J字[2013]2319号)~~

摘　　要：抑制性消减杂交(suppression subtractive hybridization,SSH)技术因操作简单、筛选效率高、假阳性率低等优点,被广泛用于生物与医学领域.该文拟找出用SSH所建立的10个肺癌差异表达基因文库中重复出现的基因,结果显示,其中6个正向杂交文库中,出现2次的基因有41个,出现3次的基因有4个;其中4个反向杂交文库中,出现2次的基因有6个.进一步用生物信息学分析发现,这些重复出现的差异表达基因参与了多种细胞功能,如基因转录、蛋白翻译、细胞粘附、DNA修复、氧化还原反应等,并涉及多条信号通路.值得注意的是,在这些差异表达基因中,核糖体蛋白相关基因占比最多.然后,用实时定量PCR检测文库中的TIMP3、GPX3、HSP90B1、CD9等基因的表达,结果显示,SSH文库中差异表达基因的上调和下调趋势具有较好的特异性.该研究为发现肺癌相关基因提供有价值的线索.Suppression subtractive hybridization（ SSH） was widely employed to construct c DNA libraries of differentially expressed genes in biology and medicine. In this study,we found out the repeated genes from the 10 lung cancer-c DNA libraries constructed by SSH and then investigated their features by bioinformatics. The results indicated that the 41 genes and 4 genes appeared respectively twice and three times in the 6 forward-subtracted libraries,while the 6 genes repeated twice in the 4 reverse-subtracted libraries. These repeated genes were involved in multiple cellular functions such as transcription,translation,cell adhesion,DNA repair,and oxidation reduction reaction,as well as several signalingpathways. Interestingly,more ribosomal protein genes were detected in the lung cancer c DNA libraries repeated genes. Furthermore,the 4 genes（ TIMP3,GPX3,HSP90B1 and CD9） were selected among the repeated genes to confirm their expression in lung cancer tissues by quantitative real-time PCR. The tendency of their up-regulation and down-regulation were in accordance with that of libraries,indicating that SSH was a specific,sensitive and powerful method to search differentially expressed genes. Our study provides valuable data for finding novel lung cancer-related genes.

关 键 词：肺癌 抑制性消减杂交 生物信息学 基因文库 

分 类 号：R734.2[医药卫生—肿瘤] Q811[医药卫生—临床医学]