Generation of Rat Monoclonal Antibody for Cytokeratin 18 by Immunization of Three-Dimensional-Cultured Cancer Cells
Generation of Rat Monoclonal Antibody for Cytokeratin 18 by Immunization of Three-Dimensional-Cultured Cancer Cells
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通过三维培养癌细胞免疫产生细胞角蛋白 18 的大鼠单克隆抗体
DOI：
10.1089/mab.2020.0030
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发表时间：
2020
影响因子：
--
通讯作者：
Yokoyama Chikako
中科院分区：
文献类型：
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作者：
Soeta Kenta;Iuchi Katsuya;Hisatomi Hisashi;Yokoyama Chikako
细胞角蛋白（CK） 18是一种中间丝蛋白，在细胞的完整性和机械稳定性中起重要的功能作用。由于CK8和CK18都是单纯上皮细胞的主要组成部分，在肿瘤的背景下，它们在大多数肿瘤中表达，并被研究作为肿瘤病理的诊断和预后标志物。CK18在细胞凋亡过程中也被一些半胱天冬酶切割。三维培养癌细胞作为玻璃癌细胞系培养和活体肿瘤之间的中间模型，在癌症研究中具有重要意义。在这项研究中，我们通过免疫3d培养的DLD-1细胞的裂解物来产生大鼠单克隆抗体（mAb），以阐明肿瘤的一个特征，我们的结果表明mAb 2H7可以识别人CK18。此外，我们发现mAb 2H7可用于免疫印迹、免疫沉淀和免疫荧光染色。因此，它可能是一个有用的诊断工具，以评估恶性肿瘤。
Cytokeratin (CK) 18 is an intermediate filament protein that plays a major functional role in the integrity and mechanical stability of cells. Since both CK8 and CK18 are major components of simple epithelia, in the context of tumors, they are expressed in most carcinomas, and have been studied as diagnostic and prognostic markers in tumor pathology. CK18 is also cleaved by some caspases during apoptosis. Three-dimensional (3D)-cultured cancer cells are useful for cancer research as an intermediate model betweenin vitrocancer cell line cultures andin vivotumors. In this study, we produced rat monoclonal antibodies (mAbs) through immunization of the lysate from 3D-cultured DLD-1 cells to elucidate a characteristic feature of a tumor, and our results showed that mAb 2H7 recognized human CK18. Furthermore, we indicated that mAb 2H7 was useful for immunoblotting, immunoprecipitation, and immunofluorescence staining. Therefore, it may be useful as a diagnostic tool for evaluating malignancy.