Institute of Immunology, People's Liberation Army, The Third Military Medical University, Chongqing, China; Department of General Surgery, Daping Hospital and Research Institute of Surgery, The Third Military Medical University, Chongqing, China.
Proteomics Clin Appl. 2007 Mar;1(3):336-42. doi: 10.1002/prca.200600718.
Patients with cancer frequently develop autoantibodies. The identification of tumor autoantigens may have utility in early cancer diagnosis and immunotherapy. In this study, we used serological proteomics analysis (SERPA) to identify tumor proteins that elicit humoral response in colorectal cancer (CRC). The CRC cell line HCT116 was used as a source of proteins for 2-DE and subsequent Western blot analysis in which individual serum from patients with CRC was analyzed for autoantibodies. An autoantibody against HSP60 identified by MS was detected in 13 out of 25 patients with CRC and 1 out of 15 healthy subjects. In addition, the HSP60 expressions in tumor tissues collected from 40 patients with CRC were assessed by immunohistochemistry, and serum specimens from 100 patients with cancer and 30 healthy controls were screened for antibody titer to HSP60 by ELISA. The results showed that expressions of HSP60 in tumor tissue and serum antibody titer to HSP60 were significantly higher in patients with CRC than in healthy subjects. Thus, we conclude that the SERPA is an excellent assay for the identification of tumor-associated antigens and tumor markers. The detection of HSP60 may have clinical utility in CRC screening, diagnosis, and immunotherapy.
癌症患者常产生自身抗体。鉴定肿瘤自身抗原可能对早期癌症诊断和免疫治疗具有应用价值。在本研究中，我们使用血清蛋白质组学分析（SERPA）来鉴定大肠癌（CRC）中引起体液免疫反应的肿瘤蛋白。CRC 细胞系 HCT116 被用作 2-DE 的来源，并用其蛋白进行后续 Western blot 分析，分析来自 CRC 患者的个体血清中的自身抗体。通过 MS 鉴定的 HSP60 自身抗体在 25 名 CRC 患者中的 13 名和 15 名健康受试者中的 1 名中被检测到。此外，通过免疫组织化学评估了从 40 名 CRC 患者收集的肿瘤组织中的 HSP60 表达，并通过 ELISA 筛查了 100 名癌症患者和 30 名健康对照者的血清标本对 HSP60 的抗体滴度。结果表明，CRC 患者肿瘤组织中的 HSP60 表达和血清 HSP60 抗体滴度均明显高于健康受试者。因此，我们得出结论，SERPA 是鉴定肿瘤相关抗原和肿瘤标志物的优秀方法。检测 HSP60 可能在 CRC 筛查、诊断和免疫治疗中具有临床应用价值。
