This work was supported by The Special Funds for Major State Basic Research of China (G1998051200).
从UniGene库中选取编号为BG231197,来自人鼻咽组织的EST序列.利用Blast检索GenBank的nr数据库和EST数据库,构建EST重叠群.利用人类基因组草图搜索法从成人正常鼻咽组织中PCR扩增获得该基因全长cDNA,命名为NAP1,GenBank登录号为AY190326.NAP1基因cDNA序列全长为573 bp,编码由85个氨基酸组成,相对分子质量为9 700的多肽.用α-32P-dCTP标记NAP1基因片段,与含15种正常成人组织的多组织RNA印迹膜杂交,结果表明NAP1基因在淋巴结和气管中高表达,转录本大小约为0.6 kb,在其他组织中不表达.NAP1蛋白质与滤泡树突状细胞(follicular dendritic cell, FDC)的一种分泌肽前体(FDC -SP)(AF435080)同源,与其他已知蛋白质无明显同源性.NAP1基因定位在染色体4q13,基因组跨越9 179 bp,含5个外显子和4个内含子.采用差异RT-PCR检测了40例经病理诊断为低分化鳞状上皮癌的鼻咽活检组织及其对侧相应部位的正常鼻咽组织中该基因的表达差异.在40例鼻咽癌中,NAP1基因表达下调的有17例(42.5%),表达上调的有6例(15%),无明显表达差异的有17例(42.5%).原位杂交和免疫组化结果显示该基因在正常鼻咽和鼻咽癌组织间质中的树突状细胞中表达,在其他间质细胞和鼻咽上皮中均不表达.以上结果表明,NAP1为树突状细胞的一种新的多肽,该基因在鼻咽癌组织中表达下调的原因,及其与鼻咽癌发生、发展的关系值得进一步探讨.
NPC-related ESTs were selected from UniGene bank. According to nr and EST database BLAST results, BG231197 was found to be a sequence that represents a new gene. The EST contig does not have complete ORF. NAP1 gene cDNA was got from normal nasopharynx tissue cDNA by PCR through searching Human Genome Draft. The Northern blot analysis revealed one transcript (0.6 kb) in lymph node and trachea. GenBank Accession number of the gene is AY190326. The length of NAP1 gene cDNA is 573 bp, ORF is from 113 bp to 370 bp. NAP1 encodes a 9 700 u protein composed of 85 amino acids. The protein resembles to FDC-SP secreted by FDC (AF435080). Bioinformatics analysis suggested that the protein is a secreted protein. NAP1 gene is located at chromosome 4q13. Genome spans 9 179 bp, containing 5 exons and 4 introns. The difference of NAP1 expression between nasopharyngeal carcinoma (NPC) and nasopharynx(NP) tissues of 40 cases was tested by differential RT-PCR. Low expression occurred in 17 NPC cases. High expression occurred in 6 NPC cases. Expression difference did not occur in 17 NPC cases. In situ hybridization (ISH) and immunohistochemistry (IH) suggested that it express in dentritic cells of NP and NPC tissues. It is meaningful to do further studies on the reason of low expression in NPC tissue and the function of NAP1 gene.
李峰,蒋卫红,杨旭宇,尹志华,冯湘玲,刘卫东,王磊,周文,姚开泰.鼻咽癌相关基因NAP1的克隆及鉴定[J].生物化学与生物物理进展,2004,31(5):464-469
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