肝癌细胞系BEL-7402与正常肝组织基因表达概况差异  

Gene expression profiles in hepatoma cell line BEL-7402 and normal liver tissues

作　　者：刘连新[1] 武林枫[1] 谢向晨[1] 姜洪池[1] 刘芝华[2] 吴旻[2] 

机构地区：[1]哈尔滨医科大学第一临床医学院普通外科,150001 [2]中国医学科学院肿瘤研究所分子肿瘤学国家重点实验室

出　　处：《中华实验外科杂志》2005年第8期930-932,共3页Chinese Journal of Experimental Surgery

基　　金：国家863计划资助项目(Z19010102);国家自然科学基金资助项目(30300339);黑龙江省青年科学基金资助项目(QC01C11)

摘　　要：目的观察肝癌细胞系的基因表达概况,寻找在肝癌细胞系及正常肝组织中差异表达基因。方法以肝癌细胞系及正常肝组织的总RNA反转录合成含有α32PdATP的cDNA为探针,与Atlas微阵列表达分析膜进行差异杂交,并应用半定量逆转录聚合酶链反应(RTPCR)和Northern杂交验证。结果放射自显影结果经AtlasImageTM软件分析显示在所分析的588种已知基因中,TFDP2、Akt1、E2F3等26个在肝癌细胞系中表达上调,TDGF1、BAK、Caspase3等22个表达下调,参与细胞增殖、凋亡、分化、细胞间相互作用、与细胞侵袭相关的基因表达水平发生了明显改变。结论通过Atlas微阵列系统分析发现的这些基因的表达改变组成了一个肝癌特异的基因表达谱,系统地研究肝癌细胞系的基因表达改变,可为肿瘤早期诊断和治疗提供线索。Objective To know a hepatoma cell line-specific gene expression profile and to identify genes that show altered expression between hepatoma cell line and normal liver tissues. Methods The cDNA probes which were labeled with α-^32P dATP were synthesized from total RNA of hepatoma cell line and corresponding almost normal liver tissues and differentially hybridized to two identical Atlas human cancer cDNA expression array membranes containing 588 known genes. A gene-specific semiquantitative reverse transcription polymerase chain reaction （RT-PCR） method and Northern blot were used to confirm the expression pattern of two known genes. Results Autoradiographic results were analyzed byspecific AtlasImage^TM（version1. 01a） software. Among the 588 genes analyzed, it was found 26 genes were up-regulated in cancer, including TFDP2, Akt1, E2F-3 etc, and 22 genes were down-regulated in cancer, including TDGF1, BAK, Caspase-3, etc. The expression levels of genes that were associated with the regulation of cell proliferation, apoptosis, differentiation, cell-cell interaction, invasion regulators and cytokines altered most. The RT-PCR and Northern blot results demonstrated that the expression pattern of two genes agreed with the results of Atlas human cancer eDNA expression array. Conclusion The results obtained from Atlas microarray provide a hepatoma cell line-specific expression profile. The differential hybridization technique of microarray may be a useful method in understanding the carcinogenesis,progression and treatment of human cancer.

关 键 词：肝癌 肿瘤细胞系 BEL-7402 肝组织 基因表达 微阵列 分子遗传学 

分 类 号：R735.7[医药卫生—肿瘤]