摘要
背景与目的:研究显示,染色体3p21是鼻咽癌高频率杂合性丢失位点,本研究在对3p21区域与鼻咽癌相关的表达序列标签(expressedsequencetag,EST)筛选的基础之上,进行鼻咽癌相关新基因的克隆和功能研究。方法:采用cDNA克隆质粒测序及RACE方法,获取新基因全长cDNA。利用因特网上核酸、蛋白质数据库及蛋白质分析软件进行生物信息学分析。通过构建pEGFP-C2/STGC3绿色荧光表达载体,与脂质体共转染COS7、CNE2细胞,进行基因表达蛋白质定位分析。用Northernblot法检测STGC3在人其他正常组织及肿瘤细胞系表达状况。结果:在3p21区域获得一个全长为1271bp的cDNA新序列,生物信息学分析与已知基因无明显同源性,属于一个新基因,编码146个氨基酸,定位于染色体3p21,被命名为STGC3(GenBank登录号:AY078383)。蛋白质定位分析显示,STGC3融合蛋白存在于细胞浆及细胞核内;MTETMArray2多组织膜Northernblot显示该基因在人正常组织及肿瘤细胞均有表达,并在淋巴瘤等多种肿瘤细胞株中阳性杂交信号弱。结论:STGC3属于一个新基因,其表达的蛋白质存在于细胞浆及细胞核内。在鼻咽癌及多种肿瘤细胞株表达下调。
BACKGROUND &OBJECTIVE: A locus of loss of heterozygosity (LOH) wit h high frequency has been found on chromosome 3p21 in nasopharyngeal carcinoma ( NPC). On the basis of our former research, this study was designed to clone and analyze a novel NPC-associated gene at this locus. METHODS: The full-length cD NA sequence of this gene was obtained by plasmid cDNA sequencing and RACE,and an alyzed by bioinformatics. The pEGFP-C2/STGC3 fusion mammalian expression vector was constructed, and transfected into COS7 and CNE2 cell lines mediated by lipo fectin to analyze the subcellular localization of gene expressing proteins. The expression of STGC3 was detected in normal tissues and tumor cell lines by North ern blot. RESULTS: An 1 271 bp full-length cDNA sequence of gene,which had no o bvious homology with other known genes in bioinformatic databases,was obtained. This gene,named STGC3 (GenBank accession number:AY078383),localized on chromosom e 3p21,and encodes a protein consisting of 146 amino acids. Protein localization analysis under fluorescent microscope indicated that STGC3 fusion protein was d istributed in nucleus and cytoplasm 24-48 hours after transfection. MTETMArray2 Northern blot analysis showed STGC3 expressed in both normal tissues and tumor cells,while its expression down-regulated in many tumor cell lines, such as Bur kitts lymphoma cell line Daudi. CONCLUSIONS: STGC3 is a novel gene, and down-regulated in NPC and many other tumor cell lines. STGC3 fusion protein distribut es in cytoplasm and nucleus.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2004年第10期1110-1115,共6页
Chinese Journal of Cancer
基金
国家自然科学基金(No.39970287No.30000188)
湖南省自然科学基金(No.03Jjy3029)~~
