摘要
目的 了解肝癌中DNA损伤修复 /DNA重组有关基因的表达概况 ,探索在肝癌与正常肝组织中的表达差异。方法 以肝癌及癌旁肝组织的总RNA反转录合成含有α 32 PdATP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RT PCR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 5 88种已知基因中 ,与DNA损伤修复 /DNA重组相关的基因共有 33个 ,其中DNA依赖蛋白激酶 (DNA PK)、DNA拓扑异构酶 (DNATOPⅠ )等 4个基因在肝癌组织中表达上调。RT PCR结果和DNATOPⅠ的Northern杂交结果 ,均证实Atlas微阵列差异杂交的准确性。结论 通过Atlas微阵列系统分析发现的这些基因表达改变 ,组成了一个肝癌特异的基因表达谱 ,是人类首次全面系统地研究肝癌的基因表达改变 ,一些与DNA损伤修复 /DNA重组相关基因的差异表达 ,为研究肝癌的发病机制和肿瘤的耐药性 ,提供了有益的线索。Atlas微阵列技术的应用 ,将为人类了解肿瘤的发生、发展和治疗提供一个较好的方法。
Objective To study the liver cancer specific DNA damage response/repair/recombination gene expression profile and the difference between liver cancer tissues and their adjacent normal tissues. Methods The cDNA probes labeled with α 32 P dATP were synthesized from total RNA of cancer tissue and adjacent liver tissues. The cDNA probes were 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 check the expression pattern of two known genes. Results Autoradiographic result analyzed by specific Atlas Image TM (version1.01a) showed among the 588 genes analyzed, 33 genes were related to damage response/repair/recombination and the expression of 4 genes including DNA dependent protein kinase, and DNA topoisomerase Ⅰ were found to be up regulated in liver cancer. The results of RT PCR and Northern blot confirmed the expression pattern of two genes which were detected by Atlas human cancer cDNA expression array. Conclusion The result obtained from Atlas microarray provides a liver cancer specific expression profile comprehensively and systematically. The findings about genes related to DNA damage response/repair/recombination may lead to understanding of the pathogenesis of liver cancer and the mechanism of drug resistant.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2001年第6期331-334,共4页
Chinese Journal of Digestion
基金
国家 8 6 3计划项目! (Z19 0 1 0 1 0 2 )
国家攀登计划项目! (18)
关键词
肝细胞癌
基因表达谱
微阵列
DNA蛋白激酶
DNA拓扑异构酶
Carcinoma, hepatocelluler
Gene expression profile
Microarray
DNA dependent protein kinase
DNA topoisomerase Ⅰ
