Remodeling of glycoconjugates on CD44 enhances cell adhesion to hyaluronate, tumor growth and metastasis in B16 melanoma cells expressing beta1,4-N-acetylglucosaminyltransferase III.
Beta1-4 N-acetylglucosaminyltransferase III (GnT-III) synthesizes bisecting N-acetylglucosamine structures on asparagine-linked oligosaccharides. Using B16-hm mouse melanoma cells stably expressing GnT-III activity as positive transfectants, the effect of bisecting N-acetylglucosamine on the function of CD44 was analyzed in association with adhesion to hyaluronate and tumor spread in mice. Transfection of GnT-III caused increased affinity of immunoprecipitated CD44 to erythro-agglutinating phytohemagglutinin, that preferentially recognizes bisecting N-acetylglucosamine, without affecting the surface CD44 amount, indicating an increase in bisecting N-acetylglucosamine residues on CD44 in positive transfectants. CD44-mediated adhesion to immobilized hyaluronate and the binding of fluorescence-labeled hyaluronate to the cell surface were increased in positive transfectants. The enhanced adhesion in positive transfectants was suppressed by the treatment with beta-N-acetylhexosaminidase, indicating that N-acetylglucosamine residues were responsible for the enhanced adhesion. Positive transfectants showed promoted CD44-mediated tumor growth and metastatic development in the spleen after subcutaneous inoculation into mice. These results indicate that glycosylation of CD44 due to GnT-III causes enhanced adhesion to hyaluronate, local tumor growth and metastatic growth in spleen, suggesting that the CD44-mediated adhesion and tumor spread can be modified through introduction of a glycosyltransferase gene.
Sheng
Y
Y
Department of Biochemistry, Osaka University Medical School, Suita City, Japan.
Yoshimura
M
M
Inoue
S
S
Oritani
K
K
Nishiura
T
T
Yoshida
H
H
Ogawa
M
M
Okajima
Y
Y
Matsuzawa
Y
Y
Taniguchi
N
N
D000117:Acetylglucosamine / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D003601:Cytotoxicity Tests, Immunologic; D005260:Female; D006001:Glycoconjugates; D006026:Glycoside Hydrolases / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000276:immunology / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000378:metabolism*; D008546:Melanoma, Experimental / Q000201:enzymology* / Q000473:pathology* / Q000556:secondary; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000378:metabolism; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D009368:Neoplasm Transplantation; D013160:Splenic Neoplasms / Q000201:enzymology / Q000473:pathology / Q000556:secondary; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D006001:Glycoconjugates; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid; D014442:Monophenol Monooxygenase; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase; D006026:Glycoside Hydrolases; D000117:Acetylglucosamine