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Blood Group ABO-related Glycosylation of Urothelial Cell Lines: Immunocytological, Enzymatic, and Genetic Characterization¹

Department of Urology, Skejby Hospital, DK-8200 Aarhus, Denmark [P. M., H. W.]; Pacific Northwest Research Foundation, Seattle, Washington 98122 [E. H. H.]; Royal Dental College, DK-2200 Copenhagen, Denmark [E. P. B., H. C.]; Department of Tumor Cell Biology, Danish Cancer Society, DK-2200 Copenhagen, Denmark [J. Z.]; and Department of Clinical Chemistry, Aarhus University Hospital, DK-8000 Aarhus C, Denmark [P. M., T. F. Ø.]

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, Le^y, Le^b, A monofucosylated types I and II, Ale^y, ALe^b, and A type III). The glycosyltransferases forming some of these epitopes (ß1-3/4 galactosyltransferase, 1–2 fucosyltransferase, 1–3 galactosyltransferase, and 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 transcibed 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.

¹ This work was supported by The Danish Cancer Society; Institute of Experimental Clinical Research, University of Aarhus; The Blood-Donor Foundation; Danish Foundation for the Advancement of Medical Science; Director Jacob Madsen's and Olga Madsen's Foundation; B. and M. Rasmussen's Memorial Grant; Clinical Oncology Trial Unit, The Danish Cancer Society, Aarhus; The Danish Cancer Research Foundation; National Cancer Institute Grant CA41521 (to E. H. H.); and funds provided by the Quest for Truth Foundation (to E. H. H.).

² To whom requests for reprints should be addressed.

Received 6/15/93. Accepted 2/23/94.

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