Proteomics-based identification of a tumor-associated antigen and its corresponding autoantibody in gastric cancer
Proteomics-based identification of a tumor-associated antigen and its corresponding autoantibody in gastric cancer
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DOI：
10.3892/or_00000719
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发表时间：
2010-04-01
期刊：
影响因子：
4.2
通讯作者：
Tanigawa, Nobuhiko
中科院分区：
文献类型：
--
作者：
Tsunemi, Soichiro;Nakanishi, Toyofumi;Tanigawa, Nobuhiko
识别新的肿瘤相关抗原和自身抗体将有助于癌症的早期诊断和更有效的免疫疗法的开发。本研究的目的是用蛋白质组学方法从胃癌细胞系MKN-1、MKN-45和KATOIII中鉴定新的肿瘤抗原，并从胃癌患者血清中鉴定相关的自身抗体。用双向凝胶电泳法对人胃癌细胞系MKN-1、MKN-45和KATOIII的蛋白质进行了分离。然后与胃癌患者、健康人和其他癌症患者的血清进行免疫印迹和抗体反应。从考马斯蓝染色的凝胶中取出阳性斑点，用基质辅助激光解吸/电离飞行时间质谱仪(MALDI-TOF/TOF MS)进行分析。胃癌患者血清出现多个斑点，MALDI-TOF/TOF MS鉴定其中一个斑点为78 kDa葡萄糖调节蛋白(GRP78)。免疫印迹显示17/60(28.3%)胃癌患者和0/20(0.0%)健康人血清中有反应。食道癌和结肠癌分别有4/15(26.7%)和3/15(20.0%)检出抗GRP78自身抗体，首次在胃癌患者中发现抗GRP78自身抗体。在这项研究中实施的蛋白质组学方法为识别可能在癌症中显示临床用途的新的血清标志物提供了一个强大的工具。
Identification of novel tumor-related antigens and autoantibodies will lead to early diagnosis of cancer and the development of more effective immunotherapies. The purpose of this study was to identify novel tumor antigens from the gastric cancer cell lines MkN-1, MkN-45 and KATOIII, and their related autoantibodies in sera of patients with, gastric cancer using a proteomics-based approach. Proteins from the gastric cancer cell lines (MkN-1, MkN-45 and KATOIII) were separated by two-dimensional polyacrylamide vel electrophoresis. followed by Western blotting and antibody reaction with sera from patients with gastric cancer, healthy individuals and patients with other cancers. Positive spots were excised from Coomassie blue stained gels and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS). Sera from patients with gastric cancer yielded multiple spots, one of which was identified as the 78 kDa glucose- regulated protein (GRP78) by MALDI-TOF/TOF MS. Western blots against recombinant GRP78 showed reactivity in sera from 17/60 (28.3%) patients with gastric cancer and 0/20 (0.0%) of healthy individuals. Autoantibodies against GRP78 were found in 4/15 (26.7%) and 3/15 (20.0%) patients with esophageal and colon cancer, respectively, We identified for the first time an autoantibody against GRP78 in gastric cancer patients. The proteomic approach implemented in this study offers a powerful tool for identifying novel serum markers that may display clinical usefulness in cancer.