舌鳞癌中高醛脱氢酶活性细胞相关基因文库的构建.docVIP

舌鳞癌中高醛脱氢酶活性细胞相关基因文库的构建 及生物信息学分析 孙守娟， 季平，杨晶 （重庆医科大学附属口腔医院颌面外科，重庆401147,） [摘要] 目的：构建舌鳞癌Tca8113细胞中高醛脱氢酶活性（ALDHhigh）细胞特异性表达基因cDNA文库并进行相应的生物信息学分析。方法：应用抑制性消减杂交(SSH)技术，以ALDHhigh细胞RNA 为测试方、低醛脱氢酶活性（ALDHlow）细胞RNA为驱动方，构建文库；应用 BLAST、Gene Set Analysis Toolkit V2等软件进行生物信息学分析。结果：随机挑选24个菌落进行PCR分析，克隆片段均匀分布在200bp～700bp，无假阳性克隆，成功构建文库；对120个克隆测序并进行BLAST分析，得到27个差异表达基因；功能聚类发现部分与细胞增殖、生长繁殖、再生等生物行为相关。结论：成功构建的ALDHhigh细胞特异性表达基因cDNA文库，为下一步研究肿瘤发生、发展机制和并为最终找到舌鳞癌干细胞特异性标志物提供帮助。 关键词 ：人舌鳞癌Tca8113； 抑制性消减杂交技术；肿瘤干细胞；醛脱氢酶；生物信息学 中图分类号：R739.86 文献标识码：A Library-Making and Bioinformatics Analysis of Highly Expressed Genes in High Aldehyde Dehydrogenase Activity Cells in Tongue Squamous Carcinoma Sun Shoujuan ,Ji Ping，Yang Jing(Dept. of Oral and Maxillofacial Surgery, Hospital of Stomatology, ChongQing MedicalUniversity, ChongQing 401147, China) [Abstract] Objective：To construct a subtracted cDNA library of specifically expressed genes of high aldehyde dehydrogenase activity (ALDHhigh) cells in human tongue squamous carcinoma Tca8113 cell line and to analyze it by bioinformatics approach. Methods: Subtractive cDNA library was established by means of suppression subtractive hybridization(SSH)on the basis of ALDHhigh cells RNA as tester and ALDHlow cells RNA as driver. Results: Random analysis of 24 clones with PCR methods showed that insert-fragments evenly distributed in 200bp～700bp range, no false positive clones, successfully constructed one SSH library. 27 differentially expressed genes were identified by BLAST. Cluster analysis indicated the involvement of the sequenced genes in regulation of proliferation，growth，reproduction. Conclusion: One subtractive cDNA library was constructed successfully with SSH ,that may provide much help for researching the mechanism of oncogenesis and finding out tongue squamous cell carcinomas stem cells-specific markers. [keywords]：uman tongue squamous carcinoma Tca8113; Suppression subtractive hybridization;Cancer stem cells; Aldehyde dehydrogenase; ；Bioinformatics Supporte