Cytokine-dependent invasiveness in B16 murine melanoma cells: Role of uPA system and MMP-9

Abstract

Abstract. 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 IFNÁ and TNF· 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 lungcolonizing potential in vivo and invasiveness through Matrigel-coated filters upon costimulation with IFNÁ and TNF·; neither IFNÁ nor TNF· 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 IFNÁ and TNF· was characterized by an enhanced uPA/uPAR and MMP-9 expression: TNF· promoted MMP-9 mRNA expression and pro-MMP-9 protein secretion, and the costimulation with IFNÁ and TNF· 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 bodyassociated form. Both monoclonal antibodies, anti-uPAR and anti-MMP-9, caused a significant reduction of invasiveness in IFNÁ/TNF·-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.

Tumoral and macrophage uPAR and MMP-9 contribute to the invasiveness of B16 murine melanoma cells

“…Aliquots of 5 ll of the cDNA were used for PCR amplification. The specific primers used for the identification of murine uPA, uPAR, MMP-2, MMP-9, b 2 -microglobulin were: uPA (sense: 0 , 258 bp product) [23]. All PCR reactions were performed using 0.1 U/ll of Go-Taq Polymerase (Promega).…”

EphA2 Reexpression Prompts Invasion of Melanoma Cells Shifting from Mesenchymal to Amoeboid-like Motility Style

“…This research work investigates the role of EphA2 in invasiveness of tumor cells, particularly its involvement as a motility factor. Previous findings of our laboratory indicate that, in the low metastatic F10-M3 melanoma cells, the proinflammatory cytokine IFN-g increases their mesenchymal type of invasiveness through up-regulation of MMPs and urokinase plasminogen activator system (14,15). F10-M3 melanoma cells represent a suitable model of tumor cells to investigate EphA2 role in cancer cell invasiveness in view of the finding that they do not express, even after inflammatory cytokine stimulation, EphA2 kinase.…”

Analysis of the Application of MMP-9 Inhibitor in Skin Melanoma: Experimental Study

“…MMP-9 is actively expressed in melanoma-B16 cells [1,2,6,14]. Therefore, selective MMP-9 inhibitor I (C 27 H 33 N 3 O 5 S; Calbiochem) at concentration of 5 nM providing selective MMP-9 inhibition was injected intramuscularly to experimental animals for 7 days starting from day 10 after tumor transplantation [3].…”