肝细胞癌高表达基因cDNA文库的构建及生物信息学分析  

Library-Making and Bioinformatics Analysis of Highly Expressed Genes in Primary Hepatocellular Carcinoma

作　　者：高英堂[1] 杜智[1] 王毅军[1] 阚学锋[1] 朱争艳[1] 景丽[1] 聂福华[1] 

机构地区：[1]天津市第三中心医院,天津300170

出　　处：《生物技术通讯》2006年第4期520-525,共6页Letters in Biotechnology

基　　金：天津市科委重点基金项目(0338010011);天津市科委基金面上项目(023616211);天津市卫生局科技基金项目(02KY01)

摘　　要：目的:构建肝细胞癌的高表达基因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7对肝癌组织和癌旁组织为实验材料,构建肝癌高表达基因的cDNA文库;使用BioEdit、BLAST和EGAD等软件进行生物信息学分析。结果:获得1450个白色阳性克隆,经PCR扩增后均有100～1000bb的插入片段,经杂交验证选取125个克隆进行测序;经BLAST、EGAD等生物信息学工具分析获得基因83个,其中细胞分裂相关基因5个,参与细胞信号转导或通讯的基因5个,参与细胞结构或运动的基因7个,细胞或器官防御相关基因11个,参与细胞内基因转录或蛋白表达的基因22个,代谢相关基因18个,另有15个未归类基因。结论:利用SSH技术可构建肝细胞癌组织差异表达基因的消减cDNA文库,为进一步深入探讨肝癌的诊断、治疗和预后评估等提供更多的分子指标。Objective： To construct a subtracted cDNA library of highly expressed genes in human primary hepatocellular carcinoma （HCC） and to analyze it by bioinformatics approaches. Methods： 7 pairs of HCC/its paracancerous tissue were used in the experiment with the suppressing subtractive hybridization（SSH）. The bioinformatics analysis was performed with BioEdit, BLAST and EGAD. Results： 1 450 white positive clones were obtained. The size of insert fragments ranged from 100 bp to 1 kb by PCR analysis. After hybridization, 125 positive clones were specially picked and sequenced. 83 genes from 125 clones analyzed by BLAST and EGAD were confirmed. They included 5 cell division related genes, 5 cell signaling/cell communication related genes, 7 cell structure/motility related genes, 11 cell/organism defense related genes, 22 gene/protein expression related genes, 18 metabolism related genes, and 15 unclassified genes. Conclusion： A subtracted cDNA library in HCC tissue may be constructed successfully with SSH. It also may provide more molecule guideline for further research of HCC＇s diagnosis, treatment and prognosis.

关 键 词：肝细胞癌 抑制性消减杂交 CDNA文库 生物信息学 

分 类 号：Q75[生物学—分子生物学] Q78