Cytokine-dependent invasiveness in B16 murine melanoma cells: role of uPA system and MMP-9.
Proteases are crucial for the spread of cancer cells from a primary tumor to the site of secondary growth. This study examined the ability of IFNgamma and TNFalpha to stimulate a better invasiveness in B16 murine melanoma cells, and investigated whether this enhanced ability was related to a higher expression of protease activities, such as urokinase plasminogen activator (uPA) and its receptor (uPAR), and matrix metalloproteinases 2 and 9 (MMP-2, MMP-9). We found that murine melanoma cells enhanced their lung-colonizing potential in vivo and invasiveness through Matrigel-coated filters upon costimulation with IFNgamma and TNFalpha; neither IFNgamma nor TNFalpha alone, at the dose used in the experiments, was able to elicit a change in the invasive/metastatic efficiency of melanoma cells. The invasive phenotype of murine melanoma cells stimulated with IFNgamma and TNFalpha was characterized by an enhanced uPA/uPAR and MMP-9 expression: TNFalpha promoted MMP-9 mRNA expression and pro-MMP-9 protein secretion, and the costimulation with IFNgamma and TNFalpha was required to potentiate the expression of mRNA and protein for uPAR, and to induce a redistribution of uPA from the soluble to the cell body-associated form. Both monoclonal antibodies, anti-uPAR and anti-MMP-9, caused a significant reduction of invasiveness in IFNgamma/TNFalpha-stimulated melanoma cells. These results indicate that invasiveness in B16 murine melanoma cells can be regulated in a cytokine-specific fashion and is dependent on the synergism between the uPA/uPAR system and MMP-9.
Bianchini
Francesca
F
Department of Experimental Pathology and Oncology, University of Florence, Florence, Italy.
D'Alessio
Silvia
S
Fibbi
Gabriella
G
Del Rosso
Mario
M
Calorini
Lido
L
D000818:Animals; D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects*; D016207:Cytokines / Q000494:pharmacology*; D004357:Drug Synergism; D007371:Interferon-gamma / Q000494:pharmacology; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D008546:Melanoma, Experimental / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009361:Neoplasm Invasiveness; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics / Q000378:metabolism
D016207:Cytokines; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D014568:Urokinase-Type Plasminogen Activator; D020780:Matrix Metalloproteinase 9