Secretory glycoconjugates of a mucin-synthesizing human colonic adenocarcinoma cell line. Analysis using double labeling with lectins.-猫眼课题宝

合成粘蛋白的人结肠腺癌细胞系的分泌糖缀合物。

DOI：

10.1007/bf00266394

发表时间：

1990

期刊：

Histochemistry

影响因子：

通讯作者：

中科院分区：

文献类型：

作者：

Phillips,TE;Frisch,EB

凝集素被用来表征由表现杯状细胞表型的人结肠腺癌来源的细胞系合成的粘蛋白糖蛋白和其他分泌型糖偶联物。尽管HT29-18N_2(N_2)细胞是克隆性来源的，但就像正常的原位杯状细胞一样，它们的粘蛋白糖基化是异质性的。只有识别N-乙酰氨基葡萄糖和唾液酸残基的麦芽凝集素和与N-乙酰氨基葡萄糖结合的琥珀酰化麦芽凝集素染色了所有N_2杯状细胞的分泌颗粒的含量。半乳糖结合凝集素白扁豆和甘氨酸分别对20%和21%的N2杯状细胞染色，而半乳糖结合凝集素蓖麻I对67%的N2杯状细胞染色，而另一种半乳糖结合凝集素Bandeiraea Simicifolia I的染色仅为19%。花生凝集素是一种凝集素，其半乳糖(β1-3)GalNAc结合部位不存在于正常结肠产生的粘蛋白上，但存在于大多数癌症结肠上皮的粘蛋白上，其染色细胞的比例为68%。四种识别糖的凝集素(豆凝集素、豌豆凝集素、四季豆凝集素E、菜豆凝集素L)通常只存在被染成38%的N_2杯状细胞的Inn连接的寡糖。这些凝集素的结合表明，在杯状细胞分泌颗粒内，粘蛋白蛋白骨架上既存在N-连接和N-连接的寡糖链，也存在非粘蛋白N-连接的糖蛋白和N-连接的粘蛋白。

Lectins were used to characterize mucin glycoproteins and other secretory glycoconjugates synthesized by a human colon adenocarcinoma-derived cell line which expresses a goblet cell phenotype. Despite being clonally derived, HT29-18N2 (N2) cells, like normal goblet cells in situ were heterogeneous in their glycosylation of mucin. Only wheat-germ agglutinin, which recognizesN-acetylglucosamine and sialic acid residues, and succinylated wheat-germ agglutinin, which bindsN-acetylglucosamine, stained the contents of all secretory granules in all N2 goblet cells. TheN-acetylgalactosamine binding lectinsDolichos biflorusandGlycine maxstained 20% and 21% of N2 goblet cells respectively.Ricinus communis I, a galactose-binding lectin, stained 67% of N2 goblet cells although staining by another galactose-binding lectin,Bandeiraea simplicifolia I, was limited to 19%. Peanut agglutinin, a lectin whose Gal(β1–3)GalNAc binding site is not present on mucins produced in the normal colon but which is found on most mucins of cancerous colonic epithelia, stained 68% of the cells.Ulex europeus I, a fucose-binding lectin, did not stain any N2 goblet cells. Four lectins (Lens culinaris, Pisum sativum, Phaseolus vulgaris E, Phaseolus vulgaris L) which recognize sugars normally present only inN-linked oligosaccharides stained up to 38% of N2 goblet cells. The binding of these lectins indicates either bothO-linked andN-linked oligosac-charide chains are present on the mucin protein backbone or the co-existence of non-mucinN-linked glycoproteins andO-linked mucins within the goblet cell secretory granule.