Altered N-glycosylation in macrophage x melanoma fusion hybrids.
It was recently reported that a majority of hybrids generated in vitro between weakly metastatic mouse Cloudman S91 melanoma cells and human or mouse macrophages showed enhanced metastatic potential (Rachkovsky et al., 1998). With few exceptions, hybrids with enhanced metastatic potential also had elevated basal melanin content, enhanced chemotactic responses to fibroblast-conditioned media, and stronger responsiveness to MSH compared to parental cells. Analyses revealed that altered N-glycosylation in metastatic hybrids could explain the multiple phenotypic changes. Tyrosinase, TRP-2 and LAMP-1 from hybrids migrated more slowly on gels compared to the same proteins from parental melanoma cells, consistent with increased glycosylation. Migration of LAMP-1 from hybrids was similar to that from peritoneal macrophages which also appeared to be more heavily glycosylated than LAMP-1 from Cloudman cells. The incorporation of 3H-glucosamine, as a marker of N-glycosylation, into tyrosinase and LAMP-1 was found to be elevated in hybrids, suppressed by N-glycosylation inhibitors and stimulated by MSH to a greater degree in hybrids compared to parental cells. These results indicate N-glycosylation as an important regulatory pathway for MSH-induced melanogenesis, and further suggest that altered N-linked glycosylation may be an underlying mechanism for regulation of both melanogenesis and metastasis in macrophage x melanoma hybrids.
Pawelek
J M
JM
Dept. of Dermatology, Yale University School of Medicine, New Haven, CT 06520, USA. john.pawelek@yale.edu
Chakraborty
A K
AK
Rachkovsky
M L
ML
Orlow
S J
SJ
Bolognia
J L
JL
Sodi
S A
SA
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D015703:Antigens, CD / Q000302:isolation & purification / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D006822:Hybrid Cells / Q000187:drug effects / Q000378:metabolism*; D019746:Intramolecular Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism; D051907:Lysosomal Membrane Proteins; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism; D008543:Melanins / Q000096:biosynthesis*; D008544:Melanocytes / Q000166:cytology / Q000378:metabolism*; D008546:Melanoma, Experimental / Q000473:pathology*; D020460:Melanosomes / Q000378:metabolism / Q000648:ultrastructure; D008562:Membrane Glycoproteins / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D014442:Monophenol Monooxygenase / Q000302:isolation & purification / Q000378:metabolism; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000378:metabolism; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000378:metabolism*; D010088:Oxidoreductases; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D011506:Proteins / Q000378:metabolism; D014407:Tumor Cells, Cultured; D000521:alpha-MSH / Q000494:pharmacology
D015703:Antigens, CD; D051907:Lysosomal Membrane Proteins; D008543:Melanins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D011506:Proteins; D000521:alpha-MSH; D010088:Oxidoreductases; C489799:TYRP1 protein, human; C079621:Tyrp1 protein, mouse; C064056:tyrosinase-related protein-1; D014442:Monophenol Monooxygenase; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase; D005944:Glucosamine; D015056:1-Methyl-3-isobutylxanthine