Measurement of antibodies against tumor suppressor gene products in health screening for lung cancer

为了建立肺癌早期诊断的血清标志物，我们发展了检测抗肿瘤抑制基因产物自身抗体的方法。此外，我们还进行了一项大规模调查，以证实这些标记物在健康人群中检测肺癌的性能。首先，我们建立了一种针对细菌产生的GST-Rb融合蛋白的免疫印迹试验。用该方法检测了45例肺癌患者和30例健康志愿者的血清。我们发现13.3%的肺癌患者血清中有抗GST-Rb融合蛋白抗体，而30名正常志愿者中没有这些抗体。其次，我们建立了检测抗Rb抗体的酶联免疫吸附试验(EL ISA)，方便了大量样品的检测。该系统能检测肺癌患者血清中升高的抗Rb抗体，但不能检测到正常人血清中的抗Rb抗体。这些结果表明我们的抗Rb抗体测量系统是有用的。第三，为了研究这些测量系统在肺癌健康筛查中的表现，我们获得了鸟取县大学伦理委员会和鸟取县地方自治机构的批准。之后，我们开始招募健康志愿者，他们在集体健康体检环境中提供他们的血清和疾病经历的信息。我们计划将这项研究继续进行三年以上。综上所述，在这笔拨款的支持下，我们制作了两种灵敏的方法来检测抗肿瘤抑制基因产物的自身抗体，验证了它们作为肿瘤标志物在肺癌患者中的有效性，并开始了大规模调查，以确认它们在肺癌健康筛查中的表现。我们相信，这将导致一种新的血清指标的开发，用于肺癌的早期检测。

To establish serum markers for early diagnosis of lung cancer, we developed methods to detect autoantibodies against tumor suppressor gene products. In addition, we conducted a large scale survey to confirm the performance of these markers to detect lung cancer among healthy people. First, we developed an immunoblotting assay against bacterially generated GST-RB fusion proteins. Using this assay, we examined 45 sera from lung cancer patients and 30 healthy volunteers. We found 13.3% of lung cancer patients had serum antibodies against GST-Rb fusion proteins, while none of 30 normal volunteers had these antibodies. Secondly, we developed Enzyme-Linked Immunosorbent Assay (ELISA) for detecting anti-Rb antibodies to facilitate measurement a lot of samples. This ELISA system could detect elevated anti-Rb antibodies in serum of lung cancer patients, but not in normal volunteers. These results indicated the usefulness of our measuring systems for anti-Rb antibodies. Thirdly, to study the performance of these measuring systems in health screening for lung cancer, we got the approval of the ethical committee of Tottori University, and of local self-governing bodies in Tottori prefecture. After that, we started to recruit healthy volunteers who provide their serum and information of disease experience in group health examination settings. We are planning to continue this study for more three years. In summary, supported by this grant, we made two sensitive methods to detect autoantibodies against tumor suppressor gene products, verified their usefulness in lung cancer patients as tumor makers, and started mass survey to confirm their performance in health screening for lung cancer. We believe this will lead to the development of a new serum indicator for early detection of lung cancer.