Petryniak J, Varani J, Ervin P R, Goldstein I J
Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor 48109-0606.
Cancer Lett. 1991 Oct;60(1):59-65. doi: 10.1016/0304-3835(91)90049-n.
Cell surface glycoproteins were isolated from the lysates of 125I-labeled normal human mammary epithelial cells (NHMEC) and from the human breast carcinoma cell line MCF-7, of blood-group O phenotype, by affinity chromatography on Griffonia simplicifolia I lectin-Sepharose. Specific elution of glycoproteins from the column with methyl alpha-D-galactoside suggests the presence of alpha-D-galactosyl groups on these moieties. SDS-PAGE analysis of isolated glycoproteins revealed both quantitative and qualitative differences between glycoproteins from normal and malignant cells. Three major glycoproteins of Mr 180 kDa, 85 kDa and the 44 kDa were obtained from MCF-7 cells. The 180-kDa glycoprotein was absent in NHMEC and the 44-kDa glycoprotein was very weakly expressed in these cells. The only glycoprotein which was found in almost equal amount in the lysate from both normal and malignant cells was the 85-kDa glycoprotein. These results indicate differences between normal human mammary epithelial cells and one kind of malignant human mammary epithelial cells, in the expression of glycoproteins containing alpha-D-galactosyl groups, irrespective of blood-group phenotype; they also demonstrate that alpha-D-galactosyl group are expressed in a very restrictive manner on the surface of this tumor cell line.
通过用西非单叶豆凝集素-Sepharose进行亲和层析,从125I标记的正常人类乳腺上皮细胞(NHMEC)以及血型为O型的人乳腺癌细胞系MCF-7的裂解物中分离细胞表面糖蛋白。用α-D-甲基半乳糖苷从柱上特异性洗脱糖蛋白,表明这些部分存在α-D-半乳糖基。对分离出的糖蛋白进行SDS-PAGE分析,揭示了正常细胞和恶性细胞来源的糖蛋白在数量和质量上的差异。从MCF-7细胞中获得了三种主要的糖蛋白,分子量分别为180 kDa、85 kDa和44 kDa。180 kDa的糖蛋白在NHMEC中不存在,44 kDa的糖蛋白在这些细胞中表达非常微弱。在正常细胞和恶性细胞裂解物中几乎等量存在的唯一糖蛋白是85 kDa的糖蛋白。这些结果表明,正常人类乳腺上皮细胞和一种恶性人类乳腺上皮细胞在含α-D-半乳糖基的糖蛋白表达上存在差异,与血型表型无关;它们还表明,α-D-半乳糖基在这种肿瘤细胞系表面以非常受限的方式表达。