应用SEREX技术筛选与鉴定鼻咽癌相关抗原  

SCREENING AND IDENTIFICATION OF NASOPHARYNGEAL CARCINOMA ASSOCIATED ANTIGENS BY SEREX

作　　者：黄绍明[1] 何少健[1] 莫发荣[1] 梁皓[2] 廖红[1] 谢小薰[1] 罗国容[1] 

机构地区：[1]广西医科大学人体解剖与组织胚胎学教研室,南宁530021 [2]广西医科大学第一附属医院眼科

出　　处：《广西医科大学学报》2005年第2期166-169,共4页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.30160024);广西壮族自治区科委基金资助项目(桂科攻No.20380212)

摘　　要：目的:应用SEREX技术筛选与鉴定鼻咽癌相关抗原,为鼻咽癌的早期诊断、免疫治疗和预后监测提供候选的生物学指标和理论基础。方法:从鼻咽癌新鲜活检组织中提取mRNA,构建cDNA表达文库。用鼻咽癌患者的血清筛选所构建的cDNA表达文库,获得的阳性克隆经扩增后提取噬菌粒DNA进行测序。所得序列与GenBank数据库中已知基因进行同源比较,分析其生物学特性。结果:所构建的cDNA原始文库含1×106个重组体,蓝白斑试验显示重组率为93%。用鼻咽癌血清对表达文库进行筛选,共获得21个阳性克隆,分别代表14个不同的cDNA插入片段。12个插入片段的DNA序列分别与基因库上的基因有不同程度的同源性,其中6个基因与鼻咽癌或其它肿瘤的发生、发展关系密切;另6个基因的生理功能未见文献报道。2个插入片段未发现有同源基因,推测可能为新发现的基因。结论:SEREX技术是目前筛选肿瘤抗原最简便、有效的手段之一。本文研究发现的抗原将为鼻咽癌的研究提供进一步的理论基础。Objective:The nasopharyngeal carcinoma (NPC) associated antigens were screened and identified by SEREX technique in order to supply biological markers for early diagnosis, immunotherapy and survival surveillance of NPC.Methods:mRNA was extracted from fresh biopsy tissues of NPC, and then the NPC cDNA expression library was constructed. The NPC library was screened by sera of NPC patients and isolated the positive clones. After amplified positive clones, undergo in vivo excision with ExAssist helper phage, the plasmid DNA was extracted and sequenced. DNA sequences were comparied with Gene Bank database by the BlAST software to analyze its homology and biological function.Result:The primary library consisted of 1×10~6 recombinants. The blue and white test showed that the recombinant rate of the cDNA library was 93%. Approximately 1×10~6 recombinants clones of NPC cDNA library were screened by the pool of sera from NPC patients at a 1∶1 000 dilution. In total, 21 positive clones were obtained that represented 14 distinct cDNA insert fragments. 6 of 14 nucleotide sequences were homologized to known gene products in Gene Bank database and close related to the occurrence and development of NPC and other tumors; another 6 of 14 nucleotide sequences were also homologized to known gene products, but their biological function are not published yet. However, 2 of 14 nucleotide sequences were not found any homological gene from GeneBank database indicated that they were novel genes.Conclusion:The SEREX technique is one of the most effective and convenient means to seek tumor antigens. The antigens discovery by us will supply helpful information for NPC research.

关 键 词：鼻咽癌 CDNA文库 SEREX 肿瘤抗原 

分 类 号：R739.62[医药卫生—肿瘤]