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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: To isolate a novel gene associated with nasopharyngeal carcinoma near the site of D6S1581 (6q25.3-27), which is located in the long arm of nasopharyngeal carcinoma of the long arm, on chromosome 6. Methods: The expression sequences of 11 expressed sequence tags (ESTs) located in the vicinity of D6S1581 were detected by differential RT-PCR in normal human nasopharyngeal epithelial cells and nasopharyngeal carcinoma cell lines. The expression of one of the ESTs in up-regulated Nasopharyngeal carcinoma and normal nasopharyngeal tissue expression analysis. Northern blotting was used to verify the difference in expression and to detect the expression in multiple organs and the size of the transcript of the gene. Using bioinformatics resources to clone and obtain its full-length cDNA, a preliminary analysis of the sequence. Results: A novel gene with up-regulated expression in nasopharyngeal carcinoma at 6q25.3-27 was obtained and named as NAG23 (FBXO30) (GenBank accession number: AF248640). The gene was up-regulated in 68.9% of nasopharyngeal carcinoma biopsies. The full-length cDNA was 3301bp and predicted that its open reading frame encoded a 390 amino acid cytoplasmic protein containing a F-box motif. Conclusion: The NAG23 gene is a novel gene that is up-regulated in NPC and may encode a new F-box protein. NAG23 may be involved in the occurrence and development of nasopharyngeal carcinoma.