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培养的大鼠非实质肝上皮细胞中的角蛋白14蛋白:角蛋白14和角蛋白19作为常用小鼠单克隆抗体OV-6抗原的特性

Keratin 14 protein in cultured nonparenchymal rat hepatic epithelial cells: characterization of keratin 14 and keratin 19 as antigens for the commonly used mouse monoclonal antibody OV-6.

作者信息

Bisgaard H C, Parmelee D C, Dunsford H A, Sechi S, Thorgeirsson S S

机构信息

Laboratory of Experimental Carcinogenesis, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-0037.

出版信息

Mol Carcinog. 1993;7(1):60-6. doi: 10.1002/mc.2940070110.

Abstract

We have recently reported that cell lines of nonparenchymal origin isolated from rat liver and pancreas, which have been suggested to be the progeny of a facultative stem cell compartment in vivo, express an unusual combination of keratins (K). These cell lines express K8 and K14 but not K18 and K5, their normal partners in filament formation (Bisgaard HC, Thorgeirsson SS, J Cell Physiol 147:333-343, 1991). However, upon spontaneous transformation and differentiation toward a hepatoblastlike progeny, K14 expression is abrogated and replaced by expression of K18 (Wirth et al., Electrophoresis 13:305-332, 1992). In the study presented here, we confirmed by protein sequence analysis that K14 was a major component of the intermediate filaments in a nonparenchymal cell line of hepatic origin. Immunocytochemical analysis of the cells in monolayer demonstrated that K8 as well as K14 were incorporated in the cellular cytoskeleton. Further analysis by immunoprecipitation showed that filament complexes were formed between K8 and K14 as atypical partners. Thus, we concluded that in some nonparenchymal cell lines isolated from rat liver, K8 and K14 form a major intermediate filament network. Finally, we showed that an antibody widely used in studies of the cell lineages of hepatic and pancreatic tissues and their neoplasms, the mouse monoclonal antibody OV-6, recognizes a common epitope in K14 and K19.

摘要

我们最近报道,从大鼠肝脏和胰腺分离出的非实质来源的细胞系,有人认为它们是体内兼性干细胞区室的后代,表达一种不寻常的角蛋白(K)组合。这些细胞系表达K8和K14,但不表达K18和K5,而K18和K5是它们在丝状物形成中的正常伙伴(Bisgaard HC,Thorgeirsson SS,《细胞生理学杂志》147:333 - 343,1991)。然而,在自发转化并向肝母细胞样后代分化时,K14的表达被消除,取而代之的是K18的表达(Wirth等人,《电泳》13:305 - 332,1992)。在本文所呈现的研究中,我们通过蛋白质序列分析证实,K14是肝源性非实质细胞系中间丝的主要成分。对单层细胞的免疫细胞化学分析表明,K8以及K14都被纳入细胞细胞骨架中。通过免疫沉淀的进一步分析表明,K8和K14作为非典型伙伴形成了丝状物复合物。因此,我们得出结论,在从大鼠肝脏分离出的一些非实质细胞系中,K8和K14形成了主要的中间丝网络。最后,我们表明,一种在肝脏和胰腺组织及其肿瘤的细胞谱系研究中广泛使用的抗体,即小鼠单克隆抗体OV - 6,识别K14和K19中的一个共同表位。

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