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肝细胞癌高表达基因cDNA文库的构建及生物信息学分析 被引量：2

Library-Making and Bioinformatics Analysis of Highly Expressed Genes in Primary Hepatocellular Carcinoma
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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.
作者 高英堂 杜智 王毅军 阚学锋 朱争艳 景丽 聂福华
机构地区 天津市第三中心医院
出处 《生物技术通讯》 CAS 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)
关键词 肝细胞癌 抑制性消减杂交 CDNA文库 生物信息学 hepatocellular carcinoma suppressing subtractive hybridization eDNA library bioinformatics
分类号 Q75 [生物学—分子生物学] Q78 [生物学—分子生物学]
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同被引文献25

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生物技术通讯
2006年 第4期

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