肝细胞癌高表达基因cDNA文库的构建及生物信息学分析

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。

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

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.