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培养的人脑肿瘤细胞上表皮生长因子受体及相关糖蛋白的表达

Expression of epidermal growth factor receptor and associated glycoprotein on cultured human brain tumor cells.

作者信息

Steck P A, Gallick G E, Maxwell S A, Kloetzer W S, Arlinghaus R B, Moser R P, Gutterman J U, Yung W K

出版信息

J Cell Biochem. 1986;32(1):1-10. doi: 10.1002/jcb.240320102.

Abstract

The expression of epidermal growth factor (EGF-R) in normal glial and glioma cells grown in culture was examined by using several independent assays. Immunoprecipitation with the monoclonal antibody R1 of extracts from metabolically labeled glial and glioma cells revealed a protein of Mr approximately 170,000, with a migration in sodium dodecyl sulfate-polyacrylamide gels identical to the EGR-R of A431 epidermal carcinoma cells. Furthermore, in the majority of glioma extracts, a protein of Mr approximately 190,000 was specifically immunoprecipitated by this antibody. Similar results were obtained by immunoblotting with a second antibody directed against a synthetic peptide in the sequence of the v-erb-B oncogene. In cell lines expressing both proteins, each was specifically phosphorylated on tyrosine in immune complex kinase assays. The majority of glioma cells bound between 40,000 to 80,000 125I-labeled epidermal growth factor molecules per cell. These results suggest that the expression of EGF-R is common in cultured human glioma cells. In addition, a structurally related protein, is expressed in some of these cells.

摘要

通过几种独立的检测方法,对培养的正常神经胶质细胞和胶质瘤细胞中表皮生长因子(EGF-R)的表达进行了检测。用代谢标记的神经胶质细胞和胶质瘤细胞提取物的单克隆抗体R1进行免疫沉淀,发现一种分子量约为170,000的蛋白质,其在十二烷基硫酸钠-聚丙烯酰胺凝胶中的迁移情况与A431表皮癌细胞的EGR-R相同。此外,在大多数胶质瘤提取物中,这种抗体能特异性免疫沉淀一种分子量约为190,000的蛋白质。用针对v-erb-B癌基因序列中合成肽的第二种抗体进行免疫印迹,也得到了类似的结果。在表达这两种蛋白质的细胞系中,每种蛋白质在免疫复合物激酶检测中酪氨酸都被特异性磷酸化。大多数胶质瘤细胞每个细胞结合40,000至80,000个125I标记的表皮生长因子分子。这些结果表明,EGF-R的表达在培养的人胶质瘤细胞中很常见。此外,一种结构相关的蛋白质在其中一些细胞中表达。

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