·289·中国肺癌杂志2009年4月第12卷第4期Chin J Lung Cancer, April 2009, Vol.12, No.4·基础研究·本研究受北京市科技新星（No.2006B34）、教育部出国留学回国启动基金和北京市优秀人才培养基金（No.20061D03）资助作者单位：101149 北京，北京市结核病胸部肿瘤研究所，北京市肿瘤分子生物学实验室肺癌分室（细胞生物学研究室）（通讯作者：岳文涛，E-mail: yuewt2000@yahoo.com）应用SEREX技术筛选和鉴定非小细胞肺癌肿瘤相关抗原岳文涛 张丽娜 郑华 王玥 孙颖 焦振峰【摘要】 背景与目的 目前用于非小细胞肺癌诊断的标志物为数不多，为寻找更多的早期诊断和靶向治疗相关的标志物，本研究应用SEREX技术筛选与鉴定非小细胞肺癌肿瘤相关抗原。方法 使用非小细胞肺癌患者血清对人肺鳞癌和腺癌噬菌体展示文库进行生物淘选和SEREX筛选。PCR扩增阳性克隆插入片段后测序，结果与GeneBank数据库中已知基因进行同源性比较，分析其生物学特性。结果 用非小细胞肺癌血清对噬菌体展示文库进行筛选，共获得31个阳性克隆，测序后发现其代表15个基因，其中12个与已知cDNA序列同源，与肺癌或其它肿瘤的发生、发展关系密切；另外3个未发现有同源基因，可能是新基因。结论 应用SEREX技术发现15个肺癌相关抗原和3个肺癌相关抗原的候选基因，为进一步的深入研究打下良好的基础。【关键词】 肺肿瘤；SEREX；噬菌体展示；基因文库【中图分类号】 R734.2 DOI:10.3779/j.issn.1009-3419.2009.04.06Screening and Identification of Non-Small-Cell Lung Cancer Associated Antigen with SEREXWentao YUE, Lina ZHANG, Hua ZHENG, Yue WANG, Ying SUN, Zhenfeng JIAODepartment of Cellular and Molecular Biology, Beijing TB and Thoracic Tumor Research Institute, Beijing 101149, ChinaCorresponding author: Wentao YUE, E-mail: yuewt2000@yahoo.com【Abstract】 Background and objective Currently, there are only limited number of diagnostic tumor markersfor non-small-cell lung cancer (NSCLC). The aim of this study is to screen and identify non-small-cell lung cancer asso-ciated antigens, by using of cancer sera containing antibodies which react with autologous cellular antigens (tumor-asso-ciated antigens TAAs), which can be used in NSCLC early screening or target treatment. Methods Two T7 phage displaycDNA libraries were panning against the sera from 24 patients with NSCLC. Then the enriched libraries were screenedwith the pool of sera from NSCLC patients, the positive clones encoding antigenic genes were obtained after screening, andthe nucleotide sequences of positive were identified and analyzed with BLAST software in GenBank. Results After im-munoscreen two T7 phage display cDNA libraries with serum from patients with NSCLC, thirty-one positive clones wereobtained and sequence results showed they were derive from 15 genes, 12 of 15 genes were homologous to the genes knownin GenBank, such as RHOA, ITGB4, and HNRNPA2/B1 et al . The other three genes expressed sequence tags (ESTs) werenot found in GenBank. Conclusion Fifteen NSCLC associated antigen genes and three candiated gene were obtained bySEREX from T7 phage display cDNA libraries.【Key words】 Lung neoplasms; SEREX; Phage display; Gene libraryThis study was supported by grants from the Beijing Novel Program (to Wentao YUE)(No.2006B34), and ScientificResearch Foundation for the Returned Overseas Chinese Scholars, State Education Ministry and Beijing Research Foun-dation for Excellent Talents(No.20061D03).非小细胞肺癌（NSCLC）生物学特性十分复杂、恶性程度高，80%的患者在确诊时已属中晚期，且发病率和死亡率呈逐年上升趋势[1]。尽管人们对NSCLC的诊断及治疗有了很大的提高，但NSCLC仍是严重威胁人们健康和生命的疾病。近年来，有效寻找肿瘤相关抗原已成为肿瘤早期诊断和肿瘤免疫治疗的研究热点。几种非小细胞肺癌标志已从血清中分离出来，如神经元特异性烯醇化酶（NSE）、胃泌素释放肽前体（pro-GRP）、细胞角蛋白19（CYFRA21-1）、p53抗体和癌胚抗原（CEA）等，但灵敏度和特异度并不高。相较于血清中肿瘤抗原型标志物，血

应用SEREX技术筛选和鉴定非小细胞肺癌肿瘤相关抗原