Selectin ligands on human melanoma cells.
Twelve established human melanoma lines were screened for surface expression of the carbohydrate antigens Lewisa (Lea), sialyl Lewisa (SLea), dimeric sialyl Lewisa (diSLea), sialyl LewisX (SLex) and dimeric sialyl LewisX (diSLeX). None of the lines expressed SLex, but 11/12 were positive for diSLeX and 7/12 were positive for SLea. Although both diSLeX and SLea have been reported to bind to E-selectin, none of the melanoma lines exhibited E-selectin-dependent adhesion to activated human umbilical vein endothelial cells (HUVECs). Three melanoma lines infected with a retroviral vector carrying the cDNA for the human Lewis fucosyltransferase (FucT-III) subsequently expressed SLeX at their cell surface and exhibited E-selectin-dependent adhesion to activated HUVECs. Treatment of these transduced cells with inhibitors of O-linked or N-linked protein glycosylation significantly inhibited E-selectin-mediated adhesion, though fluorescence-activated cell sorter analysis indicated no decrease in cell surface expression of SLeX, Slea or diSLeX. This suggests that the majority of SLeX/SLea-type glycans endogenously produced by human melanoma cells are not protein-associated and do not mediate E-selectin-dependent adhesion. These results support the hypothesis that E-selectin-dependent adhesion requires presentation of SLeX-type moieties on appropriate glycoproteins.
Miller
N
N
Imperial Cancer Research Fund, Rayne Institute, St Thomas' Hospital, London, UK.
Vile
R G
RG
Hart
I R
IR
D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D018076:DNA, Complementary / Q000235:genetics; D019040:E-Selectin / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism; D005647:Fucosyltransferases / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D016256:Lewis X Antigen / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D008545:Melanoma / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D000080506:Sialyl Lewis X Antigen; D014162:Transfection; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D019040:E-Selectin; D016256:Lewis X Antigen; D008024:Ligands; D009844:Oligosaccharides; D000080506:Sialyl Lewis X Antigen; D005647:Fucosyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase