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通过细胞学标记、亲和色谱法和新糖蛋白靶向光解分析人视网膜母细胞瘤和视网膜母细胞系的内源性糖受体(凝集素)谱。

Endogenous sugar receptor (lectin) profiles of human retinoblastoma and retinoblast cell lines analyzed by cytological markers, affinity chromatography and neoglycoprotein-targeted photolysis.

作者信息

Griegel S, Rajewsky M F, Ciesiolka T, Gabius H J

机构信息

Institut für Zellbiologie (Tumorforschung), Universität Essen, Federal Republic of Germany.

出版信息

Anticancer Res. 1989 May-Jun;9(3):723-30.

PMID:2764517
Abstract

Plant lectins have previously been employed to map the composition of cellular glycoconjugates on retinoblastoma and other tumor cells. To characterize the cellular receptors in protein-carbohydrate interactions, we have applied cytological markers (fluorescent neoglycoproteins) containing common carbohydrate building-blocks to investigate the presence of endogenous carbohydrate-binding proteins in six human retinoblastoma cell lines and one established human retinoblast cell line. The staining patterns showed similar expression of endogenous sugar receptors in all cell lines, with few qualitative differences. However, application of affinity chromatography using resins with immobilized carbohydrates as affinity ligands to isolate sugar receptors (lectins) with binding specificities for beta-D-galactosides, alpha-D-mannosides, alpha-L-fucosides alpha-D-glucosides and to N-acetyl-D-glucosamine and N-acetyl-D-galactosamine, respectively, revealed significant differences between the cell lines, emphasizing the value of complementary biochemical analysis. To demonstrate the practical use of this type of glycobiochemical profiling, selective photodestruction of retinoblastoma cells in vitro was accomplished following incubation with synthetic neoglycoprotein-hematoporphyrin conjugates and subsequent exposure to light. This phototherapeutical approach thus combined the inherent specificity of a neoglycoprotein for a particular cellular phenotype with targeted drug activation.

摘要

植物凝集素先前已被用于绘制视网膜母细胞瘤和其他肿瘤细胞上细胞糖缀合物的组成。为了表征蛋白质 - 碳水化合物相互作用中的细胞受体,我们应用了含有常见碳水化合物构建块的细胞学标记(荧光新糖蛋白),以研究六种人视网膜母细胞瘤细胞系和一种已建立的人视网膜母细胞系中内源性碳水化合物结合蛋白的存在。染色模式显示所有细胞系中内源性糖受体的表达相似,在质量上差异很小。然而,使用固定化碳水化合物作为亲和配体的树脂进行亲和色谱,以分别分离对β - D - 半乳糖苷、α - D - 甘露糖苷、α - L - 岩藻糖苷、α - D - 葡萄糖苷以及对N - 乙酰 - D - 葡萄糖胺和N - 乙酰 - D - 半乳糖胺具有结合特异性的糖受体(凝集素),结果显示细胞系之间存在显著差异,强调了互补生化分析的价值。为了证明这种糖生物化学分析类型的实际应用,在与合成新糖蛋白 - 血卟啉缀合物孵育并随后暴露于光之后,在体外实现了视网膜母细胞瘤细胞的选择性光破坏。因此,这种光疗方法将新糖蛋白对特定细胞表型的固有特异性与靶向药物激活相结合。

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