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Cell Growth & Differentiation Vol. 12, 623-630, December 2001
© 2001 American Association for Cancer Research

Fusion Hybrids with Macrophage and Melanoma Cells Up-Regulate N-Acetylglucosaminyltransferase V, ß1–6 Branching, and Metastasis1

Ashok K. Chakraborty2, John Pawelek, Yoshitaka Ikeda, Eiji Miyoshi2, Natalia Kolesnikova, Yoko Funasaka, Masamitsu Ichihashi and Naoyuki Taniguchi

Department of Dermatology, Yale University School of Medicine, New Haven, Connecticut 06510 [A. K. C., J. P., N. K.]; Department of Biochemistry, Osaka University Medical School, Osaka 565-0871, Japan [Y. I., E. M., N. T.]; and Department of Dermatology, Kobe University School of Medicine, Kobe 650-0017, Japan [Y. F., M. I.]

It was shown previously that a majority of hybrids produced by in vitro fusion of normal macrophages with Cloudman S91 melanoma cells displayed enhanced metastatic potential in vivo, increased motility in vitro, increased ability to produce melanin, and responsiveness to melanocyte stimulating hormone compared with the parental Cloudman S91 melanoma cells. These hybrids also showed altered N-glycosylation consistent with a slower migration pattern of lysosome-associated membrane protein (LAMP-1) on electrophoretic gels. Because LAMP-1 is the major carrier of polylactosamine sugar structures, and synthesis of this complex sugar moiety indicates the extent of ß1,6 branch formation by ß1,6-N-acetyl-glucosaminyltransferase V (GnT-V), we analyzed the expression of GnT-V and ß1,6 branching in highly metastatic macrophage-fusion hybrids and compared with poorly metastatic ones. GnT-V was up-regulated in regard to both mRNA levels and enzymatic activity specifically in metastatic hybrids as well as parental macrophages compared with weakly metastatic hybrids and parental melanoma cells. Macrophages and metastatic hybrids also showed increased binding of the lectin L-phytohemagglutinin, which specifically binds to the ß1,6-branched sugar moiety. In addition, in metastatic hybrids there was increased cell surface expression of LAMP-1 and ß1 integrin, two prominent substrates for GnT-V also known to be associated with metastasis. Finally, exposure of metastatic hybrids in vitro to L-phytohemagglutinin or LAMP-1 completely eliminated melanocyte stimulating hormone/ isobutylmethyl xanthine-induced motility, suggesting a role for GnT-V in the motility of these cells. In summary, macrophage fusion with melanoma cells often increased metastatic potential, which was associated with enhanced expression of GnT-V and ß1,6-branching in glycoproteins. It is suggested that the known correlation with elevated GnT-V in both human and animal metastasis could, at least in some cases, reflect previous fusion of tumor cells with tumor-infiltrating macrophages, which, similar to malignant cells, show elevated expression of GnT-V and ß1,6-branched polylactosamines.




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Copyright © 2001 by the American Association of Cancer Research.