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投稿时间：2018-09-10 网络发布日期：2019-06-20
投稿时间：2018-09-10 网络发布日期：2019-06-20
中文摘要: 目的 筛选肺癌抗原组中强免疫原性抗原,以期为肺癌的早期诊断、疫苗研制提供参考。方法 选取5份异体肺癌患者血清以重组cDNA表达文库血清学分析(SEREX)技术筛选肺癌cDNA表达文库,并进行生物信息学分析。此外,选取2017年2月至2018年2月收治的肺癌患者60例为肺癌组,另取同期进行体检的正常人员60例为对照组。分别采集两组人员血清标本,对比相关肺癌抗原的异体血清反应结果。并对差异有统计学意义的肺癌抗原进行受试者工作特征(ROC)曲线分析,明确其在肺癌早期诊断中的价值。结果 本实验共完成3轮血清学筛选,最终得到70个阳性克隆。采用NCBI网上序列分析软件BLAST对各cDNA序列的同源性进行比对,结果显示上述70个阳性克隆分别表示63个不同基因,包括35个已知基因、26个功能未知基因以及2个新基因。其中35个已知基因中仅有DNA拓扑异构酶Ⅱα(Top2A)基因与肺癌相关,2个新基因分别为热休克蛋白90α(HSP90α)、真核翻译延长因子1γ(EF-1γ),功能未知基因中可能与肺癌相关的包括二羟基苯甲酸内置结合蛋白1配偶体(POB1)、中枢神经系肽酶Ⅰ(TPP1)，共5种肺癌相关抗原。肺癌组Top2A、HSP90α、POB1抗原克隆的IgG血清抗体阳性率高于对照组(P<0.01)。Top2A、HSP90α、POB1抗原克隆联合检测诊断肺癌的曲线下面积、敏感性、特异性高于上述三项指标单独检测(P<0.05)。结论 SEREX技术可用于肺癌抗原组中强免疫原性抗原的筛选、鉴定,临床工作中可将Top2A、HSP90α、POB1基因作为肺癌早期诊断以及免疫治疗的靶向基因。
中文关键词: 肺癌 重组cDNA表达文库,血清学分析技术 免疫性抗原 筛选 免疫治疗
Abstract:ObjectiveTo screen strong immunogenic antigen in lung cancer antigen group and provide guidance for the early diagnosis and vaccine development of lung cancer. Methods Five serum samples from allogeneic lung cancer patients were collected to screen lung cancer cDNA expression library by recombinant cDNA expression library (SEREX) technology,and bioinformatics analysis was carried out.In addition,60 patients with lung cancer treated from February 2017 to February 2018 were designed as lung cancer group,and 60 normal persons receiving physical examination at the same period were served as control group.Serum samples were collected respectively for antigen cloning in two groups and compared with the results of foreign serum responses of 5 lung cancer antigens.In addition,receiver operating characteristic (ROC) curve of lung cancer antigens with statistical significance was analyzed to clarify their value in the early diagnosis of lung cancer. Results Three rounds of serological screening were completed ，and 70 positive clones were obtained.Using NCBI online sequence analysis software BLAST,the homology comparison of each cDNA sequence showed that the 70 positive clones represented 63 different genes,including 35 known genes,26 unknown functional genes and 2 new genes.Among the 35 known genes,only DNA topoisomerase Ⅱ alpha (Top2A) gene was associated with lung cancer,2 new genes were heat shock protein 90 alpha (HSP90α) and eukaryotic translation elongation factor 1 gam (EF-1γ).Among functionally unknown genes,dihydroxybenzoic acid-binding protein 1 (POB1) and central nervous system peptidase I (TPP1) were related to lung cancer.The positive rates of IgG serum antibody of Top2A,HSP90α and POB1 antigenclones in lung cancer group were statistically higher than those in control group (P<0.01).The area under curve,sensitivity and specificity of combined detection of Top2A,HSP90α and POB1 antigen clones were statistically higher than those of individual detection of Top2A,HSP90α and POB1 in the diagnosis of lung cancer (P<0.05). Conclusion SEREX technology can be used to screen and identify strong immunogenic antigens in lung cancer antigens.Top2A,HSP90α and POB1 can be used as target genes for early diagnosis and immunotherapy of lung cancer in clinic.
keywords: Lung cancer Recombinant cDNA expression library Immunological antigen Screening Immunotherapy
文章编号： 中图分类号： 文献标志码：A
基金项目:延安市科学技术研究发展计划项目(2016KS-09)
附件
| Author Name | Affiliation |
| LIU Dong-li,HE Xiao-long,DU Yan-ling,YANG Shuan-ying,PENG Dan | Department of Respiratory Medicine,Affiliated Hospital of Yan′an University,Yan′an,Shaanxi 716000,China |
引用文本：
刘东利,贺小龙,杜延玲,杨拴盈,彭丹.肺癌抗原组中强免疫原性抗原的筛选[J].中国临床研究,2019,32(6):751-754.
刘东利,贺小龙,杜延玲,杨拴盈,彭丹.肺癌抗原组中强免疫原性抗原的筛选[J].中国临床研究,2019,32(6):751-754.