肺癌抗原组中强免疫原性抗原的筛选
|
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。
Objective To 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(Top2 A) 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 Top2 A,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 Top2 A,HSP90α and POB1 antigen clones were statistically higher than those of individual detection of Top2 A,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.Top2 A,HSP90α and POB1 can be used as target genes for early diagnosis and immunotherapy of lung cancer in clinic.
Objective To 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(Top2 A) 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 Top2 A,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 Top2 A,HSP90α and POB1 antigen clones were statistically higher than those of individual detection of Top2 A,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.Top2 A,HSP90α and POB1 can be used as target genes for early diagnosis and immunotherapy of lung cancer in clinic.
引文
[1] Tsutsumi K,Matsuya Y,Sugahara T,et al.Inorganic polyphosphate enhances radio-sensitivity in a human non-small cell lung cancer cell line,H1299[J].Tumour Biol,2017,39(6):1010428317705033.
[2] 孙楠,孙守国,臧若川,等.ENO1自身抗体联合癌胚抗原在肺腺癌诊断中的临床应用[J].中华检验医学杂志,2018,41(6):446-449.
[3] 杨昌恒,庹必英,廖江荣,等.支气管肺泡灌洗液及血清中肿瘤标志物在肺癌诊断中的临床应用[J].中华肺部疾病杂志(电子版),2018,11(3):334-335.
[4] 刘昀,孙秀珍,徐晶,等.法国梧桐花粉变应原cDNA表达文库的构建和初步鉴定[J].山西医科大学学报,2015,46(4):327-330
[5] 何晓锋,肖越勇.肿瘤冷冻免疫及联合细胞免疫治疗的现状与进展[J].中华放射学杂志,2013,47(4):381-384.
[6] Nakao M,Muramatsu H,Kagawa Y,et al.Immunological status may predict response to nivolumab in non-small cell lung cancer without driver mutations[J].Anticancer Res,2017,37(7):3781-3786.
[7] 魏智民,陈寅,焦顺昌.免疫检查点抑制剂在肺癌治疗中的应用研究进展[J].中华老年多器官疾病杂志,2018,17(5):383-388.
[8] 陈若冰,陈涛.疫苗在肺癌治疗中的研究进展[J].中国全科医学,2018,21(23):2777-2790.
[9] 郑燕华,邹德威,冯凯,等.蛋白芯片技术筛选肝癌血清标志蛋白的初步研究[J].中华检验医学杂志,2005,28(6):628-631.
[10] 林裕辉,李旭,刘坚军,等.基于iTRAQ的定量蛋白组学分析鉴定下肢动脉硬化闭塞症潜在的血清生物标记物[J].中华实验外科杂志,2014,31(12):2820-2822.
[11] 袁娜,辛国宏,左笑笑,等.联合应用噬菌体展示技术与重组cDNA表达文库血清学分析技术筛选肺癌早期诊断相关抗原[J].浙江大学学报(医学版),2014,43(4):388-396.
[12] 杨军,刘妮,李宗芳,等.一种新的组学技术-肿瘤抗原组学[J].现代肿瘤医学,2013,21(6):1375-1378.
[13] 韩正祥,张梦瑾,张英楠,等.TOP2A 在非小细胞肺癌中的高表达促进肿瘤细胞的增殖和侵袭能力[J].现代肿瘤医学,2016,24(9):1371-1375.
[14] 王传新.肺癌早期诊断的新肿瘤标志研究[J].中华临床实验室管理电子杂志,2017,5(4):212-216.
[15] 张晶锐,高峰,李荣江.HSP90α、MMP-9在胆管癌组织中的表达及其临床意义[J].中国临床研究,2017,30(8):1067-1069.
[16] 王鹏,闫平平,刘会娟,等.肺癌候选抗原的血清反应分析及其联合检测的应用[J].第四军医大学学报,2009,30(21):2278-2280.
[17] Tan Y,Wang J,Su S,et al.Enhancement of endocytic uptake of HIV-1 virions into CD4-negative epithelial cells by HIV-1 gp41 via its interaction with POB1[J].Cell Mol Immunol,2017,14(6):568-571.
[2] 孙楠,孙守国,臧若川,等.ENO1自身抗体联合癌胚抗原在肺腺癌诊断中的临床应用[J].中华检验医学杂志,2018,41(6):446-449.
[3] 杨昌恒,庹必英,廖江荣,等.支气管肺泡灌洗液及血清中肿瘤标志物在肺癌诊断中的临床应用[J].中华肺部疾病杂志(电子版),2018,11(3):334-335.
[4] 刘昀,孙秀珍,徐晶,等.法国梧桐花粉变应原cDNA表达文库的构建和初步鉴定[J].山西医科大学学报,2015,46(4):327-330
[5] 何晓锋,肖越勇.肿瘤冷冻免疫及联合细胞免疫治疗的现状与进展[J].中华放射学杂志,2013,47(4):381-384.
[6] Nakao M,Muramatsu H,Kagawa Y,et al.Immunological status may predict response to nivolumab in non-small cell lung cancer without driver mutations[J].Anticancer Res,2017,37(7):3781-3786.
[7] 魏智民,陈寅,焦顺昌.免疫检查点抑制剂在肺癌治疗中的应用研究进展[J].中华老年多器官疾病杂志,2018,17(5):383-388.
[8] 陈若冰,陈涛.疫苗在肺癌治疗中的研究进展[J].中国全科医学,2018,21(23):2777-2790.
[9] 郑燕华,邹德威,冯凯,等.蛋白芯片技术筛选肝癌血清标志蛋白的初步研究[J].中华检验医学杂志,2005,28(6):628-631.
[10] 林裕辉,李旭,刘坚军,等.基于iTRAQ的定量蛋白组学分析鉴定下肢动脉硬化闭塞症潜在的血清生物标记物[J].中华实验外科杂志,2014,31(12):2820-2822.
[11] 袁娜,辛国宏,左笑笑,等.联合应用噬菌体展示技术与重组cDNA表达文库血清学分析技术筛选肺癌早期诊断相关抗原[J].浙江大学学报(医学版),2014,43(4):388-396.
[12] 杨军,刘妮,李宗芳,等.一种新的组学技术-肿瘤抗原组学[J].现代肿瘤医学,2013,21(6):1375-1378.
[13] 韩正祥,张梦瑾,张英楠,等.TOP2A 在非小细胞肺癌中的高表达促进肿瘤细胞的增殖和侵袭能力[J].现代肿瘤医学,2016,24(9):1371-1375.
[14] 王传新.肺癌早期诊断的新肿瘤标志研究[J].中华临床实验室管理电子杂志,2017,5(4):212-216.
[15] 张晶锐,高峰,李荣江.HSP90α、MMP-9在胆管癌组织中的表达及其临床意义[J].中国临床研究,2017,30(8):1067-1069.
[16] 王鹏,闫平平,刘会娟,等.肺癌候选抗原的血清反应分析及其联合检测的应用[J].第四军医大学学报,2009,30(21):2278-2280.
[17] Tan Y,Wang J,Su S,et al.Enhancement of endocytic uptake of HIV-1 virions into CD4-negative epithelial cells by HIV-1 gp41 via its interaction with POB1[J].Cell Mol Immunol,2017,14(6):568-571.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |