Tumor cell surface beta 1-4-linked galactose binds to lectin(s) on microvascular endothelial cells and contributes to organ colonization.
Cell surface carbohydrate structures acting as ligands for tissue specific mammalian lectins have been implicated in cell-cell interactions during embryogenesis, lymphocyte homing, and tumor cell metastasis. In this report, we provide evidence that beta 1-4 linked galactose (Gal) residues in N-linked oligosaccharides on the surface of blood born tumor cells serve as a ligand for binding to microvascular endothelial cells. D36W25, a class 1 glycosylation mutant of the MDAY-D2 lymphoreticular tumor cell line, lacks sialic acid and Gal in cellular glycans due to a defect in the Golgi UDP-Gal transporter. Using UDP-Gal and bovine galactosyltransferase in vitro, beta 1-4 Gal was restored to the surface of the cells and 70% of the galactosylated glycans persisted for 8 h in vitro at 37 degrees C. Compared to mock-treated D36W25 cells, galactosylated D36W25 cells showed an 80% increase in binding to microvascular endothelial cell monolayers in vitro. The enhanced binding of galactosylated D36W25 cells to endothelial cell was inhibited by the addition of lactosamine-conjugated albumin to the assay. Consistent with these observations, swainsonine and castinospermine, two inhibitors of N-linked processing that result in loss of lactosamine antennae inhibited the binding of wild-type MDAY-D2 cells to endothelial cells in vitro. Injection of radiolabeled tumor cells into the circulation of syngeneic mice, showed that galactosylation of D36W25 cells resulted in 2-3 more tumor cells retained in the lungs and livers. In addition, galactosylation of D36W25 cells increased by 30-fold the number of visible liver metastases on inspection 4 wk after tumor cell injection. These results suggest that beta 1-4Gal-binding lectins on microvascular endothelial cells can contribute to retention and secondary tumor formation of blood born tumor cells. With the increasing availability of purified glycosyltransferases, reconstruction of a variety of carbohydrate sequences on the surface of class 1 mutants provides a controlled means of studying carbohydrate-lectin interactions on viable cells.
Cornil
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I
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario.
Kerbel
R S
RS
Dennis
J W
JW
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005434:Flow Cytometry; D005690:Galactose; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors; D007212:Indolizines; D007700:Kinetics; D037102:Lectins; D008228:Lymphoma, Non-Hodgkin; D051379:Mice; D008811:Mice, Inbred DBA; D009154:Mutation; D009844:Oligosaccharides; D017026:Swainsonine; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000502:physiology*; D054836:Indolizidines
D000470:Alkaloids; D005690:Galactose; D006026:Glycoside Hydrolases; D007212:Indolizines; D037102:Lectins; D009844:Oligosaccharides; D017026:Swainsonine; C037806:castanospermine; D054836:Indolizidines