鼻咽癌组织差异表达cDNA序列的克隆与鉴定  

Cloning and Identification for Differentially Expressed cDNA in Nasopharyngeal Carcinoma Tissue

作　　者：马英红[1] 王秀清 曾木圣[1] 李冰[3] 曾益新[1] 

机构地区：[1]中山医科大学肿瘤防治中心肿瘤研究所,广东广州510060 [2]深圳市人民医院(暨南大学第二附属医院),广东深圳518020 [3]广州医学院,广东广州510182

出　　处：《癌症》2001年第1期18-22,共5页Chinese Journal of Cancer

基　　金："973"重点项目<‘疾病基因组学"理论和技术体系建立>项目;中山医科大学"211工程"重点项目

摘　　要：目的：分析鼻咽癌组织中的差异基因，寻找与鼻咽癌病理过程相关的新的癌基因和抑癌基因。方法：应用抑制性消减杂交法，构建了鼻咽组织与正常鼻咽组织的消减库；用差异筛选技术筛选出了阳性克隆；用Ｓｏｕｔｈｅｒｎ　ｂｌｏｔ和 Ｎｏｒｔｈｅｒｎ　ｂｌｏｔ杂交证实了阳性克隆的可靠性；用序列分析确定了阳性克隆的性质。结果：从消减库中获得了１２个阳性克隆；序列分析表明，８个克隆与已知基因高度同源，２个为新的候选基因，２个新的候选基因在两个不同的消减库中确实存在差异，而其中的一个新基因在鼻咽癌组织和细胞系中下调表达。结论：抑制性消减杂交是分析相关基因的较好的方法，发现了两个与鼻咽癌病理过程有关的新的ｃＤＮＡ。Objective: The current study was designed to analyze the differentially expressed genes in nasopharyngeal carcinoma (NPC) tissue and identify the novel oncogenes and the tumor suppressor genes associated with NPC pathogenesis. Metholls: Using the suppression subtractive hybridization (SSH), two SSH pools (normal nasopharyngeal tissue cDNA-pool and NPC tissue cDNA-pool) were constructed. Positive clones were screened by differential screen technique, and conformed using Southern blot and Northern blot, and sequenced with ABI 377 DNA sequencer. Results: Twelve positive clones were identified from SSH pools. Sequences of the 12 positive clones showed that & clones were homologous with the genes published in GenBank and 2 clones were unknown genes. The two novel genes NPS and NP9 in the normal control cDNA-pool were richer than that in the cancerous cDNA-pool and NP9 was further found to be down-regulated in both NPC cell lines and NPC tissue. Conclusions: The SSH is useful for analysis of the disease-related genes and two novel cDNAs were identified to be related with NPC pathogenesis.

关 键 词：鼻咽肿瘤 癌基因 抑癌基因 消减文库 CDNA 克隆 

分 类 号：R739.63[医药卫生—肿瘤] R730.23[医药卫生—临床医学]