Pattern expression of glycan residues in AZT-treated K562 cells analyzed by lectin cytochemistry.
The present paper shows that human chronic myeloid (K562) cells exposed 3 h to 20 microM 3'-azido-3'-deoxythymidine (AZT) exhibit marked variations of the oligosaccharide moiety of glycoconjugates. These changes were analyzed by confocal fluorescence microscopy, upon incubation of control and AZT-treated cells with biotin-lectin conjugates to visualize cell surface glycans or total glycans after cells permeabilization. In addition, cell fluorescence distribution of the biotinylated lectins, localized with streptavidin conjugates labeled with Alexa Fluor 488, was analyzed by flow cytometry. The results obtained show significant variations on the expression/distribution of membrane surface glycans as detected by both WGA and SNA, two lectins that recognize primarily cellular internal membrane glycolipids. A further interesting result was the significant increase of N-acetylglucosamine linked glycans localized either at the cell surface or intracellularly but only in K562 cells exposed to AZT. On the whole, our data demonstrate that AZT alters both lipid and N-linked glycosylations thus confirming previous observations, from our laboratory and from other Authors, that the drug impair the nucleotide-sugar import in the Golgi's lumen. AZT does also alter the O-linked glycosylations that occur in the Golgi complex since these reactions require the incorporation of sialic acid, GlcNAc and GalNAc all of which are sensitive to the drug.
Lizzi
Anna Rita
AR
Department of Biomedical Sciences and Technologies, University of L'Aquila, 67100 L'Aquila, Italy.
D'Alessandro
Anna Maria
AM
Bozzi
Argante
A
Cinque
Benedetta
B
Oratore
Arduino
A
D'Andrea
Gabriele
G
D019921:Biotinylation; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005453:Fluorescence; D005456:Fluorescent Dyes; D006651:Histocytochemistry; D006801:Humans; D020014:K562 Cells; D037102:Lectins / Q000378:metabolism*; D018613:Microscopy, Confocal; D010539:Permeability / Q000187:drug effects; D011134:Polysaccharides / Q000378:metabolism*; D015215:Zidovudine / Q000494:pharmacology*
D005456:Fluorescent Dyes; D037102:Lectins; D011134:Polysaccharides; D015215:Zidovudine