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目的 分离位于染色体 3p2 4- 2 6区域中鼻咽癌相关的新基因。方法 在染色体 3p2 4- 2 6鼻咽癌杂合性丢失 (loss of heterozygosity,L OH)高频区 ,用 RT- PCR及 Northern杂交 ,检测了 2 0个表达序列标记 (expressed sequence tag,EST)在鼻咽癌细胞株 HNE1和原代培养的正常鼻咽上皮细胞中的表达水平 ,并对其中一个在鼻咽癌细胞株 HNE1中表达下调的 EST,在 7个正常鼻咽活检组织和 19例鼻咽癌活检组织中的表达进行了检测。用 c DNA文库筛选方法获得其全长 c DNA序列 ,对该序列进行了初步分析。结果 获得了一个位于染色体 3p2 5 .3的新基因 ,命名为 NAG- 7基因 (Gen Bank收录编号 :AF0 86 70 9)。该基因在 2 6 .3% (5 / 19)的鼻咽癌活检组织中表达下调 ,全长 16 6 7bp,其开放阅读框 (open reading frame,ORF)编码一个含 94个氨基酸的碱性蛋白质 ,相对分子质量为 110 2 3870 ,预测为一跨膜蛋白质。它含有一个蛋白激酶 C磷酸化位点和肉豆蔻基位点。迄今为止 ,NAG- 7基因序列在基因库中未见任何同源性基因。结论 NAG- 7基因是一个在鼻咽癌中表达下调的新基因 ,它的改变可能参与了鼻咽癌的发生发展。
Objective To isolate a novel gene associated with nasopharyngeal carcinoma in the 3p2 4- 2 6 region of chromosome. METHODS: Twenty expressed sequence tags (ESTs) were detected by RT-PCR and Northern blot in the high frequency region of chromosome 3p2 4- 2 6 loss of heterozygosity (LOH) In nasopharyngeal carcinoma cell line HNE1 and primary cultured normal nasopharyngeal epithelial cells, and one of the ESTs down-regulated in nasopharyngeal carcinoma cell line HNE1 was detected in 7 normal nasopharyngeal biopsy tissues and 19 Nasopharyngeal carcinoma biopsy tissue expression was tested. The full length c DNA sequence was obtained by screening with c DNA library, and the sequence was analyzed preliminarily. As a result, a new gene located on chromosome 3p2 5 .3 was obtained and named as NAG-7 gene (Gen Bank Accession No. AF0 86 70 9). The gene was down-regulated in 26.3% (5/19) NPC tissues with a total length of 16 6 7bp. The open reading frame (ORF) gene encoded a 94 amino acid basic protein , The relative molecular mass of 110 2 3870, predicted as a transmembrane protein. It contains a protein kinase C phosphorylation site and a myristyl site. To date, no homology gene has been found in the gene pool for the NAG-7 gene sequence. Conclusion NAG-7 gene is a new gene whose expression is downregulated in nasopharyngeal carcinoma. Its alteration may be involved in the development of nasopharyngeal carcinoma.