Gao Hongjun, Zheng Min, Sun Sijin, Wang Hongwu, Yue Zhigang, Zhu Yun, Han Xiaochen, Yang Junquan, Zhou Yanqiu, Cai Yiran, Hu Wanning
Department of Clinical Laboratory, China Meitan General Hospital, Beijing, PR China.
Department of Oncology, Tangshan People's Hospital & Tangshan Cancer Hospital, North China University of Science and Technology, Tangshan, PR China.
Oncotarget. 2017 Jul 18;8(38):64170-64179. doi: 10.18632/oncotarget.19369. eCollection 2017 Sep 8.
Novel tumor antigens and their related autoantibodies have tremendous potential for early diagnosis of non-small cell lung cancer (NSCLC). In this study, we identify antigens from NSCLC tissue and autoantibodies in sera of patients with NSCLC using a modified proteomics-based approach. We seperated and identified four NSCLC-associated proteins extracted from the cytosol in tumor tissues by mini-two-dimensional gel electrophoresis, followed by Western blot and hybridization with individual sera for confirmation of antibody binding. Of the proteins we identified, we selected 58 kDa chaperonin containing TCP1(T-Complex Protein 1) subunit 5 (CCT5) for validation. Serum levels of carcinoembryonic antigen (CEA) and cytokeratin 19 fragments (CYFRA 21-1) were measured in all serum samples with an immunoluminometric assay and a receiver operating characteristic (ROC) curve was analyzed for autoantibodies against CCT5, CEA and CYFRA 21-1. The results show that CCT5 can induce an autoantibody response in NSCLC sera and show higher expression in NSCLC tissues by immunohistochemistry and Western blot. Anti-CCT5 autoantibody was found in 51% (23/45) of patients with NSCLC, but only 2.5% (1/40) in non-tumor individual controls. A receiver operating characteristic curve constructed with a panel of autoantibodies against CCT5 (AUC=0.749), CEA (AUC=0.6758), and CYFRA 21-1(AUC=0.760) show a sensitivity of 51.1% and 97.5% specificity in discriminating NSCLC from matched controls. These results indicate the potential utility of screening autoantibodies in sera, show that CCT5 could be used as a biomarker in cancer diagnosis.
新型肿瘤抗原及其相关自身抗体在非小细胞肺癌(NSCLC)的早期诊断中具有巨大潜力。在本研究中,我们采用改良的蛋白质组学方法,从NSCLC组织中鉴定抗原以及NSCLC患者血清中的自身抗体。我们通过小型二维凝胶电泳分离并鉴定了从肿瘤组织胞质溶胶中提取的四种与NSCLC相关的蛋白质,随后进行蛋白质印迹,并与个体血清杂交以确认抗体结合。在我们鉴定出的蛋白质中,我们选择了含TCP1(T-复合物蛋白1)亚基5(CCT5)的58 kDa伴侣蛋白进行验证。使用免疫发光测定法测量所有血清样本中癌胚抗原(CEA)和细胞角蛋白19片段(CYFRA 21-1)的血清水平,并针对抗CCT5、CEA和CYFRA 21-1的自身抗体分析受试者工作特征(ROC)曲线。结果表明,CCT5可在NSCLC血清中诱导自身抗体反应,并且通过免疫组织化学和蛋白质印迹显示在NSCLC组织中表达更高。在51%(23/45)的NSCLC患者中发现了抗CCT5自身抗体,但在非肿瘤个体对照中仅为2.5%(1/40)。由一组抗CCT5(AUC = 0.749)、CEA(AUC = 0.6758)和CYFRA 21-1(AUC = 0.760)自身抗体构建的受试者工作特征曲线在区分NSCLC与匹配对照时显示出51.1%的敏感性和97.5%的特异性。这些结果表明了筛查血清中自身抗体的潜在效用,表明CCT5可作为癌症诊断中的生物标志物。