The influence of N- and O-glycosylation inhibitors on the glycosylation profile of cellular membrane proteins and adhesive properties of carcinoma cell lines.
The effects of N- and O-glycosylation inhibitors on the expression of membrane proteins (MUC1 and some integrins) were evaluated in human endometrial (Ishikawa) and breast (MCF-7) cancer cells. Subconfluent cells were treated with 1-3 mg% concentration of tunicamycin and 2-10 mM of benzyl-N-acetyl-alpha-galactosaminide for 1-2 days, and used for flow cytometry, immunohistochemical staining, adhesion test and Western blotting. Benzyl-N-acetyl-alpha-galactosaminide inhibits MUC1 expression on the surface of breast more than endometrial cancer cells. Tunicamycin reduces MUC1 concentration on the cellular surface more than benzylglycoside, and greatly reduces glycosylation of glycoproteins, causing an increase in cell adhesion in both types of cancer cells. The expression of alpha2beta1 integrins on the surface of these cells was weak and decreased after treatment with inhibitors. Two different glycoforms of MUC1 proteins in endometrial cells and three in breast cancer cells were expressed and their molecular weights were reduced after treatment with glycosylation inhibitors. It was confirmed with lectin detection of carbohydrate epitopes (Tn and T) in MUC1 proteins. These observations show that glycosylation inhibitors altered the N- and O-glycan patterns in a sufficient manner, and positively modified the biological features of cancer cells.
Paszkiewicz-Gadek
Anna
A
Department of Medical Chemistry, Medical University of Bialystok, 15-230 Bialystok, Poland. zachemog@amb.edu.pl
Porowska
Halina
H
Lemancewicz
Dorota
D
Wolczynski
Slawomir
S
Gindzienski
Andrzej
A
D000951:Antigens, Neoplasm / Q000378:metabolism; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D024042:Collagen Type I / Q000378:metabolism; D016889:Endometrial Neoplasms / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005434:Flow Cytometry; D005690:Galactose / Q000031:analogs & derivatives / Q000494:pharmacology; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D038982:Integrin alpha2beta1 / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D018396:Mucin-1; D009077:Mucins / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D014415:Tunicamycin / Q000494:pharmacology
C411946:1-benzyl-2-acetamido-2-galactopyranoside; D000951:Antigens, Neoplasm; D015295:Antigens, Tumor-Associated, Carbohydrate; D024042:Collagen Type I; D038982:Integrin alpha2beta1; C060192:MUC1 protein, human; D008565:Membrane Proteins; D018396:Mucin-1; D009077:Mucins; C040286:Tn antigen; D014415:Tunicamycin; C016467:Thomsen-Friedenreich antigen; D005690:Galactose