Blood group ABO-related glycosylation of urothelial cell lines: immunocytological, enzymatic, and genetic characterization.
Three immortalized, human urothelial cell lines were characterized with respect to their ABO-related carbohydrate phenotypes using a panel of monoclonal antibodies directed to a series of carbohydrate epitopes (Lac, sialylated Lac, Le(a), sialylated Le(a), Le(x), sialylated Le(x), H types I and II, Ley, Leb, A monofucosylated types I and II, ALey, Aleb, and A type III). The glycosyltransferases forming some of these epitopes (beta 1-3/4 galactosyltransferase, alpha 1-2 fucosyltransferase, alpha 1-3 galactosyltransferase, and alpha 1-3-N-acetyl-galactosaminyltransferase) were determined by enzyme assays. The ABO gene complex was analyzed by Southern blotting, Northern blotting, and polymerase chain reaction across the O deletion and across base differences between the A and B alleles. The immunocytochemical stainings showed marked differences between the three cell lines; the high grade (tumorigenic, metastatic) cell line showed difucosylated types I and II structures, and the low grade (nontumorigenic, nonmetastatic) cell lines showed monofucosylated types I and II structures. Polymerase chain reaction genotyping of the cell lines indicated that one was OO, one was AA, and one was A plus a mutated allele. Northern blotting showed RNA encoding the A transferase. However, even though both of the A cell lines seemed to have an intact gene, which could produce A transferase and transcribed RNA, none of them showed any activity of the A gene encoded enzyme or any A-structures at the cell surface. In contrast, the three other examined glycosyltransferases were active. The three urothelial cell lines reflect in vivo findings in humans. They represent a competent system for in vitro studies of the different carbohydrate transferase genes responsible for the carbohydrate structures expressed on the cell surface in bladder tumors.
Meldgaard
P
P
Department of Urology, Skejby Hospital, Aarhus, Denmark.
Holmes
E H
EH
Bennett
E P
EP
Clausen
H
H
Zeuthen
J
J
Wolf
H
H
Orntoft
T F
TF
D000017:ABO Blood-Group System / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002295:Carcinoma, Transitional Cell / Q000737:chemistry / Q000201:enzymology*; D005838:Genotype; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D009092:Mucous Membrane / Q000737:chemistry / Q000201:enzymology; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D001743:Urinary Bladder / Q000737:chemistry / Q000201:enzymology; D001749:Urinary Bladder Neoplasms / Q000737:chemistry / Q000201:enzymology*
D000017:ABO Blood-Group System