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不同分化程度的活体人类神经母细胞瘤细胞上的寡糖。一项使用糖特异性凝集素和糖苷酶的流式细胞术研究。

Oligosaccharides on living human neuroblastoma cells of dissimilar degrees of differentiation. A flow-cytometric study with sugar-specific lectins and glycosidases.

作者信息

Pepperl B, Bohn B, Sauer A, Brossmer R

机构信息

Institut für Biochemie II (Med. Fak) der Universität Heidelberg, Germany.

出版信息

Cell Biophys. 1993 Aug-Dec;23(1-3):165-85. doi: 10.1007/BF02796511.

Abstract

The present study is an attempt to correlate cell-surface saccharide composition and/or disposition with malignant behavior and differentiation of two established human neuroblastoma sublines. The methodology applied was quantitative flow-cytometric evaluation of binding data for sugar-specific lectins in conjunction with cell-surface modification by specific glycosidases. The relevant parameters were both the number of binding sites and their apparent affinity constants for the respective lectins on native cells as well as the expected shift of those values after sequential treatment with specific glycosidases. The main conclusions from the findings may be summarized as follows: 1. There appears to exist a correlation between differentiation and/or maturation of neural cells and their cell-surface oligosaccharide patterns, as deduced indirectly by the biophysical approach of quantitative evaluation of lectin-binding data. More specifically, our findings support the hypothesis of a strong correlation between the degree of sialylation of terminal saccharide structures and the relative immaturity and/or lack of differentiation of the respective cells by morphological and biochemical criteria. 2. The combined application of specific lectins and glycosidases should be further exploited for similar purposes since it yields unequivocal information, provided that all biochemical and biophysical methods are scrutinized for their specificity. 3. Flow cytometry with fluorescence-labeled lectins is especially suited for the purposes mentioned since it allows quantitative binding studies to be conducted in a quick and uncomplicated manner. Most importantly, these data can be derived from intact living cells.

摘要

本研究旨在关联两种已建立的人神经母细胞瘤亚系的细胞表面糖组成和/或分布与恶性行为及分化情况。所应用的方法是对糖特异性凝集素的结合数据进行定量流式细胞术评估,并结合特定糖苷酶对细胞表面进行修饰。相关参数包括天然细胞上各凝集素的结合位点数量及其表观亲和常数,以及用特定糖苷酶顺序处理后这些值的预期变化。研究结果的主要结论可总结如下:1. 如通过凝集素结合数据定量评估的生物物理方法间接推断,神经细胞的分化和/或成熟与其细胞表面寡糖模式之间似乎存在关联。更具体地说,我们的研究结果支持以下假设:根据形态学和生化标准,末端糖结构的唾液酸化程度与相应细胞的相对不成熟和/或缺乏分化之间存在强关联。2. 特定凝集素和糖苷酶的联合应用应进一步用于类似目的,因为只要对所有生化和生物物理方法的特异性进行仔细审查,就能产生明确的信息。3. 用荧光标记凝集素的流式细胞术特别适合上述目的,因为它能以快速且简便的方式进行定量结合研究。最重要的是,这些数据可来自完整的活细胞。

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