Department of Biochemistry, School of Medicine, Pusan National University, Beomeo-ri, Mulgeum-eup, Yangsan 50612, Korea.
Department of Internal Medicine, Pusan National University Hospital, 1-10 Ami-dong, Seo-gu, Busan 49241, Korea.
Curr Oncol. 2021 May 20;28(3):1927-1937. doi: 10.3390/curroncol28030179.
To identify cancer/testis (CT) antigens and immunogenic proteins, immunoscreening of testicular and small-cell lung cancer cell line NCI-H889 cDNA libraries was performed using serum obtained from a small-cell lung cancer (SCLC) patient. We obtained 113 positive cDNA clones comprised of 74 different genes, designated KP-SCLC-1 through KP-SCLC-74. Of these genes, 59 genes were found to be related to cancers by EMBASE analysis. Three of these antigens, including KP-SCLC-29 (NOL4), KP-SCLC-59 (CCDC83), and KP-SCLC-69 (KIF20B), were CT antigens. RT-PCR and western blot analysis showed that NOL4 was frequently present in small-cell lung cancer cell lines (8/9, 8/9). In addition, NOL4 mRNA was weakly, or at a low frequency, or not detected in various cancer cell lines. Our results reveal that NOL4 was expressed at protein levels in small-cell lung cancer tissues (10/10) but not detected in lung adenocarcinoma and squamous cell carcinoma by immunohistochemical analysis. Serological response to NOL4 was also evaluated by western blot assay using NOL4 recombinant protein. A humoral response against NOL4 proteins was detected in 75% (33/44) of small-cell lung cancer patients and in 65% (13/20) of healthy donors by a serological western blot assay. These data suggest that NOL4 is a specific target that may be useful for diagnosis and immunotherapy in SCLC.
为了鉴定癌症/睾丸（CT）抗原和免疫原性蛋白，我们使用来自小细胞肺癌（SCLC）患者的血清对睾丸和小细胞肺癌细胞系 NCI-H889 cDNA 文库进行免疫筛选。我们获得了 113 个阳性 cDNA 克隆，由 74 个不同的基因组成，命名为 KP-SCLC-1 到 KP-SCLC-74。通过 EMBASE 分析，这些基因中有 59 个与癌症有关。其中 3 种抗原，包括 KP-SCLC-29（NOL4）、KP-SCLC-59（CCDC83）和 KP-SCLC-69（KIF20B），是 CT 抗原。RT-PCR 和 Western blot 分析显示 NOL4 频繁存在于小细胞肺癌细胞系中（8/9，8/9）。此外，NOL4mRNA 在各种癌细胞系中表达水平较低或检测不到。我们的结果表明，NOL4 在小细胞肺癌组织中表达蛋白水平（10/10），但免疫组织化学分析未在肺腺癌和鳞状细胞癌中检测到。通过 Western blot 分析用 NOL4 重组蛋白评估 NOL4 的血清学反应。血清学 Western blot 分析显示，75%（33/44）的 SCLC 患者和 65%（13/20）的健康供体存在针对 NOL4 蛋白的体液反应。这些数据表明，NOL4 是一个特异性靶标，可能对 SCLC 的诊断和免疫治疗有用。
