应用SEREX技术筛选和鉴定非小细胞肺癌肿瘤相关抗原

Screening and Identification of Non-Small-Cell Lung Cancer Associated Antigen with SEREX

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。

Background and objective Currently, there are only limited number of diagnostic tumor markers for non-small-cell lung cancer （NSCLC）. The aim of this study is to screen and identify non-small-cell lung cancer associated antigens, by using of cancer sera containing antibodies which react with autologous cellular antigens （tumor-associated antigens TAAs）, which can be used in NSCLC early screening or target treatment. Methods Two T7 phage display cDNA libraries were panning against the sera from 24 patients with NSCLC. Then the enriched libraries were screened with the pool of sera from NSCLC patients, the positive clones encoding antigenic genes were obtained after screening, and the nucleotide sequences of positive were identified and analyzed with BLAST software in GenBank. Results After immunoscreen two T7 phage display cDNA libraries with serum from patients with NSCLC, thirty-one positive clones were obtained and sequence results showed they were derive from 15 genes, 12 of 15 genes were homologous to the genes known in GenBank, such as RHOA, ITGB4, and HNRNPA2/B 1 et al. The other three genes expressed sequence tags （ESTs） were not found in GenBank. Conclusion Fifteen NSCLC associated antigen genes and three candiated gene were obtained by SEREX from T7 phage display cDNA libraries.