ULTRASTRUCTURAL-LOCALIZATION OF HELIX-POMATIA AGGLUTININ (HPA)-BINDING SITES IN HUMAN BREAST-CANCER CELL-LINES AND CHARACTERIZATION OF HPA-BINDING GLYCOPROTEINS BY WESTERN BLOTTING
ULTRASTRUCTURAL-LOCALIZATION OF HELIX-POMATIA AGGLUTININ (HPA)-BINDING SITES IN HUMAN BREAST-CANCER CELL-LINES AND CHARACTERIZATION OF HPA-BINDING GLYCOPROTEINS BY WESTERN BLOTTING
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DOI：
10.3109/01913129509014603
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发表时间：
1995-01-01
影响因子：
1
通讯作者：
SCHUMACHER, U
中科院分区：
文献类型：
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作者：
MITCHELL, BS;VERNON, K;SCHUMACHER, U
在超微结构上，5个人乳腺癌细胞系（MCF 7，BT549，BT20，T47D和HBL100）的细胞通常显示其上皮来源的许多特征。在MCF7细胞中观察到最具特色的特征，其始终显示微绒毛和膜下颗粒。这些超微结构的研究结果作为定位的基础上的凝集素Hessopomatia凝集素（HPA）的结合位点。使用预包埋方法后，在所有细胞系中获得一致的HPA标记的细胞膜，并与MCF7细胞中的微绒毛和膜下颗粒。无论使用何种方法，HBL100细胞均未被HPA标记。此外，对这些细胞系的裂解物进行聚丙烯酰胺凝胶电泳和HPA蛋白质印迹，以进一步分析这些结合位点。然而，在Western印迹中，HBL100细胞的裂解物，与所有来自其他细胞系的裂解物一样，显示了许多HPA反应性条带，表明Western印迹与组织化学制备相比具有更高的灵敏度。主带约为90 kDa，这表明这可能与转铁蛋白受体有关。
Ultrastructurally, cells of five human breast cancer cell lines (MCF7, BT549, BT20, T47D, and HBL100) generally displayed many characteristics of their epithelial origin. The most distinctive features were observed in MCF7 cells, which consistently showed microvilli and submembranous granules. These ultrastructural findings served as a basis for localizing the binding sites of the lectin Helix pomatia agglutinin (HPA). After use of the pre-embedding method consistent HPA labeling of the cell membrane was obtained in all the cell lines, and in association with microvilli and submembranous granules in the MCF7 cells. The HBL100 cells were not labeled by HPA irrespective of the method used. In addition, lysates from these cell lines were subjected to polyacrylamide gel electrophoresis and Western blotting with HPA to analyze these binding sites further. In the Western blots, however, lysates of HBL100 cells, in common with all those from the other cell lines, revealed a number of HPA-reactive bands, indicating the greater sensitivity of Western blots compared with the histochemical preparation. The principal band was of approximately 90 kDa, and it was suggested that this could be related to the transferrin receptor.