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通过定量凝集素-金超微结构细胞化学揭示野生型和培养的艾氏瘤细胞表面唾液酸糖缀合物的差异表达。

Differential expression of cell surface sialoglycoconjugates on wild-type and cultured Ehrlich tumor cells as revealed by quantitative lectin-gold ultrastructural cytochemistry.

作者信息

Roth J, Li W P, Knibbs R N, MacCallum D K, Song Z, Goldstein I J

机构信息

Department of Pathology, University of Zurich, Switzerland.

出版信息

Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11353-7. doi: 10.1073/pnas.91.24.11353.

Abstract

Three variants of the classical Ehrlich ascites tumor (EAT) cell have been studied by quantitative, sialic acid-specific, lectin-gold ultrastructural cytochemistry. Electron microscopic examination revealed pronounced differences in the surface morphology of the three cell variants. The wild-type Ehrlich cells (EAT-wt), grown in the peritoneal cavity of mice, exhibited a smooth surface profile. A variant form selected for growth as monolayer on basement membrane (EAT-c) showed a complex surface profile with numerous microvilli. The third variant (EAT-c/m), the cultured cells reinoculated into mice and passaged 20-25 times as ascites, presented a smooth surface profile similar to the EAT-wt cells. Quantitative single as well as double lectin-gold labeling revealed significant differences in the nature of cell surface sialoglycoproteins. The most significant finding was the presence of cell surface Neu5Ac alpha 2-6Gal residues as detected with the Sambucus nigra lectin on EAT-c and EAT-c/m cells, whereas EAT-wt cells contained little or none of such carbohydrate sequences. On the contrary, labeling by Maackia amurensis lectin, which recognizes the Neu5Ac alpha 2-3Gal beta 1-4GlcNAc sequence, was intense on all three Ehrlich cell variants; it was 20-60 times greater than alpha-2,6-linked sialic acid-containing glycoconjugates. Specific cell surface lectin binding combined with morphologic study appears to have identified a small subpopulation of cells within the ascites tumor that are capable of attaching to and growing on a basement membrane.

摘要

通过定量、唾液酸特异性、凝集素-金超微结构细胞化学方法,对经典艾氏腹水瘤(EAT)细胞的三种变体进行了研究。电子显微镜检查显示,这三种细胞变体的表面形态存在明显差异。在小鼠腹腔中生长的野生型艾氏细胞(EAT-wt)表面轮廓光滑。一种选择在基底膜上单层生长的变体形式(EAT-c)显示出具有许多微绒毛的复杂表面轮廓。第三种变体(EAT-c/m),即重新接种到小鼠体内并以腹水形式传代20 - 25次的培养细胞,呈现出与EAT-wt细胞相似的光滑表面轮廓。定量单凝集素和双凝集素金标记显示,细胞表面唾液酸糖蛋白的性质存在显著差异。最显著的发现是,用黑接骨木凝集素检测到EAT-c和EAT-c/m细胞表面存在Neu5Acα2 - 6Gal残基,而EAT-wt细胞几乎不含有或不含有此类碳水化合物序列。相反,识别Neu5Acα2 - 3Galβ1 - 4GlcNAc序列的黑龙江马珂蛤凝集素在所有三种艾氏细胞变体上的标记都很强;它比含α-2,6连接唾液酸的糖缀合物高20 - 60倍。特异性细胞表面凝集素结合与形态学研究相结合,似乎已经在腹水瘤中鉴定出一小部分能够附着在基底膜上并在其上生长的细胞亚群。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5990/45229/d3c7a1a9df60/pnas01146-0081-a.jpg

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