鼻咽癌细胞中表达上调的新基因NAG23(FBXO30)的克隆与分析  

Molecular Cloning and Analysis of a Novel Gene, NAG23 (FBXO 30),Upregulated in Nasopharyngeal Carcinoma

作　　者：李忠花[1] 曹利[1] 向娟娟[1] 张必成[1] 向秋[1] 唐珂[1] 周鸣[1] 李小玲[1] 李伟芳[1] 李桂源[1] 

机构地区：[1]中南大学湘雅医学院肿瘤研究所,湖南长沙410078

出　　处：《癌症》2001年第9期906-910,共5页Chinese Journal of Cancer

基　　金：国家"863"项目(项目编号:102-10-01-05);国家"973"重点项目(项目编号:G1998051008)

摘　　要：目的:分离位于6号染色体长臂鼻咽癌高频等位基因不平衡位点D6S1581(6q25.3-27)附近与鼻咽癌相关的新基因。方法:运用差异RT-PCR检测定位于D6S1581附近的11个表达序列标签(expressedsequencetags,EST)在正常人鼻咽上皮细胞和鼻咽癌细胞系中的表达水平,并对其中一个表达上调的EST在鼻咽癌组织和正常鼻咽组织中的表达情况进行分析。用Northern杂交验证其表达差异并检测其在多脏器中的表达及其所代表基因的转录本大小。利用生物信息学资源克隆并获得其全长cDNA,对该序列进行初步分析。结果:获得了一个位于6q25.3-27的在鼻咽癌中表达上调的新基因,命名为NAG23(FBXO30)(GenBank收录编号:AF248640)。该基因在68.9%的鼻咽癌活检组织中表达上调,cDNA全长3301bp,预测其开放阅读框编码一个含390个氨基酸的胞浆蛋白,该蛋白含一个F-box基序。结论:NAG23基因是一个在鼻咽癌中表达上调的新基因,很可能编码一新的F-box蛋白。NAG23可能参与了鼻咽癌的发生发展。Objective: The aim of this study was to seek novel genes associated with human nasopharyngeal carcinoma (NPC) around the microsatellite D6S1581 with high frequency of allelic imbalance in NPC tissues at the long arm of chromosome 6q25.3-27. Methods: Eleven epithelial derived expressed sequence tags (EST) were selected from the region around D6S1581. Differential RT PCR was applied to detect the expression of these EST in NPC cell line, HNE1, and the primary cultures of normal nasopharyngeal epithelial cells. Then, expression of EST AI536544 was further detected in 3 normal nasopharyngeal biopsies and 29 NPC biopsies by differential RT PCR. By Northern blots, the differential expression of the EST AI536544 was confirmed, the expression of the EST in the human eight tissues and the size of the transcript of the EST representative gene were also determined. The putative full length cDNA of the EST representative gene was cloned and the sequence of the novel gene was analyzed by bioinformatics. Results: EST AI536544 was overexpressed in the NPC cell line, HNE1, and in 68.9% of NPC biopsies. The full length cDNA with no homology to any reported genes in the database of GenBank was obtained and named NAG23 (FBXO 30)(GenBank accession number, AF248640), which is located around D6S1581(6q25.3-27).A 390 amino acid cytoplasm protein with a F box motif was deduced from the potential open reading frame of the 3301bp full length cDNA. Conclusion: NAG23 is a novel gene upregulated in NPC, which likely encode an F box protein and may be involved in development of NPC.

关 键 词：鼻咽肿瘤 基因克隆 F-BOX蛋白 NAG23基因 

分 类 号：R739.63[医药卫生—肿瘤] R730.231[医药卫生—临床医学]