Forsman L M
Department of Pathology, University of Helsinki, Finland.
Acta Pathol Microbiol Immunol Scand A. 1987 Nov;95(6):315-23. doi: 10.1111/j.1699-0463.1987.tb00047_95a.x.
The topographical distribution and characteristics of an apocrine epithelial differentiation antigen (AEA) were studied in one endometrial adenocarcinoma cell line (HEC-1-B) and two mammary carcinoma cell lines (MCF-7 and T47-D), using an antiserum raised against glycoproteins which had been isolated from human milk fat globule membranes. Immunofluorescent staining of HEC-1-B and MCF-7 cells grown in monolayers, or of histological sections of cells grown in fibrin sponges, revealed a strictly polar distribution of the antigen. The antigen was present only in the dorsal-apical cell membrane. In contrast, T47-D cells grown under identical conditions displayed a nonpolar membrane distribution of the antigen. Detergent lysates of HEC-1-B, MCF-7 and T47-D cells, surface-labeled by the PI(sodiumetaperiodate)-NaB3H4 method, were immunoprecipitated and analysed by SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis). Two closely spaced bands having an apparent MW of 315-290 kD (kilodalton) (HEC-I-B), 330-295 kD (MCF-7) and 320-270 kD (T47-D). The surface expression of the antigen was found to be hormonally regulated. Cultivation of HEC-1-B and MCF-7 in the presence of prolactin increased the amount of antigen. The T47-D cells responded only weakly to prolactin, but displayed enhanced antigen expression after treatment by estrogen and/or progesterone, as quantified by 125I protein A radioimmunoassay. The behaviour of the membrane antigen reported here provides a new and interesting marker for the differentiation and maintenance of polarity in cultured malignant cells of secretory epithelial origin.
利用针对从人乳脂肪球膜中分离出的糖蛋白制备的抗血清,研究了一种顶泌汗腺上皮分化抗原(AEA)在一种子宫内膜腺癌细胞系(HEC-1-B)和两种乳腺癌细胞系(MCF-7和T47-D)中的拓扑分布和特征。对单层培养的HEC-1-B和MCF-7细胞,或纤维蛋白海绵中生长的细胞的组织切片进行免疫荧光染色,结果显示该抗原呈严格的极性分布。该抗原仅存在于背侧顶端细胞膜中。相比之下,在相同条件下培养的T47-D细胞显示该抗原在细胞膜上呈非极性分布。采用PI(高碘酸钠)-NaB3H4法对HEC-1-B、MCF-7和T47-D细胞进行表面标记,然后用去污剂裂解,进行免疫沉淀,并通过SDS-PAGE(十二烷基硫酸钠聚丙烯酰胺凝胶电泳)分析。出现了两条紧密相邻的条带,其表观分子量分别为315 - 290 kD(千道尔顿)(HEC-I-B)、330 - 295 kD(MCF-7)和320 - 270 kD(T47-D)。发现该抗原的表面表达受激素调节。在催乳素存在的情况下培养HEC-1-B和MCF-7可增加抗原量。T47-D细胞对催乳素反应较弱,但经雌激素和/或孕酮处理后抗原表达增强,这通过125I蛋白A放射免疫测定法进行了定量分析。本文报道的膜抗原的行为为分泌上皮来源的培养恶性细胞的分化和极性维持提供了一种新的有趣的标志物。