应用免疫沉淀和蛋白质组学发现多发性骨髓瘤肿瘤相关抗原的研究  

The Discovery of Tumor Associated Antigens in Multiple Myeloma by Immunoprecipitation and Proteomics

作　　者：田恩冰[1] 黎金庆 潘书娟[1] 郭娟 赵一琳 范钦刚 王文娟[3] 杨曦明[1] TIAN Enbing;LI Jinqing;PAN Shujuan;GUO Juan;ZHAO Yilin;FAN Qingang;WANG Wenjuan;YANG Ximing(Department of Clinical Laboratory,Tongzhou District of Dongzhimen Hospital,Beijing University of Traditional Chinese Medicine,Beijing 101121,China;Oncology Department,Tongzhou District of Dongzhimen Hospital,Beijing University of Traditional Chinese Medicine,Beijing 101121,China;School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)

机构地区：[1]北京中医药大学东直门医院通州院区检验科,北京101121 [2]北京中医药大学东直门医院通州院区肿瘤科,北京101121 [3]首都医科大学中医药学院,北京100069

出　　处：《标记免疫分析与临床》2022年第6期1021-1026,共6页Labeled Immunoassays and Clinical Medicine

基　　金：国家自然科学基金面上项目(补肾生血药调控JAK2-STATs通路纠正造血细胞增殖——凋亡失衡改善化疗后骨髓抑制的机制研究)(编号:81774176)。

摘　　要：目的利用免疫沉淀和蛋白质组学发现多发性骨髓瘤的肿瘤相关抗原,探索多发性骨髓瘤的发病机制和发现新的诊断标志物。方法收集8例多发性骨髓瘤患者和8例健康对照者的血清标本并分别混合,用Protein G PLUS-Agarose免疫沉淀试剂分离和纯化多发性骨髓瘤患者和正常人血清中的抗体,并用此抗体与多发性骨髓瘤细胞系U266细胞裂解液混合,形成抗体-抗原复合物,通过1维不连续聚丙烯酰胺凝胶电泳(1D-SDS-PAGE)分离蛋白质,切胶酶解后通过液相色谱串联质谱(LC-MS/MS)分离并鉴定,用Proteome Discovery软件进行蛋白质数据库检索。结果MM组较对照组共筛选出14个上调的蛋白,7个下调的蛋白,分别列在表1和表2中。结论利用免疫沉淀及蛋白质组学筛选出了21个可能与肿瘤自身免疫相关的差异蛋白,其中的α-烯醇化酶、波形蛋白、热休克蛋白90与文献记载有较好的一致性,可作为肿瘤相关抗原的候选蛋白。Objective Using immunoprecipitation and proteomics to identify tumor associated antigens of multiple myeloma,and to explore the pathogenesis of multiple myeloma and identify new diagnostic markers.Methods Serum samples from eight MM patients and eight healthy controls were collected and mixed.Antibodies from these two groups of pooled serum samples were isolated and purified by Protein G PLUS-Agarose immunoprecipitation reagent respectively and then mixed with U266 cell lysates to isolate antibody-antigen complexes.The protein components of the complexes were separated by 1D-SDS-PAGE and protein gel bands were excised then digested with trypsin and analyzed by LC-MS/MS followed by protein database searching with Proteome Discovery software.Results Proteomic analysis identified 14 elevated proteins and 7 down-regulated proteins which were listed in Table 1 and Table 2.Conclusion By immunoprecipitation and proteomics,21 differential proteins that may be related to tumor autoimmunity are screened out.Among them,α-enolase,vimentin and heat shock protein 90 are in good agreement with previous literatures,and could be used as new candidate proteins for tumor associated antigens.

关 键 词：多发性骨髓瘤 肿瘤相关抗原 α-烯醇化酶 波形蛋白 热休克蛋白90 

分 类 号：R733.3[医药卫生—肿瘤]