In vitro and in vivo antimetastatic effect of a penicillin derivative in murine melanoma cells

BARRIONUEVO E; CAYROL MF; CREMASCHI GA; CORNIER P.G.; DELPICCOLO CM; BOGGIAN D; MATA E.G.; ROGUIN, L. P; BLANK VC

Mar del Plata

Congreso; LXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica, LI Reunión Anual de la Asociación Argentina de Farmacología Experimental, XXI Reunión Anual de la Sociedad Argentina de Biología, XXXI Reunión Anual de la Sociedad Argentina de Parasit; 2019

SAIC, SAI, SAFE

In a previous work, we demonstrated that TAP7f, an antitumor penicillin derivative formed by penicillin linked to the dipeptide Leu-Phe through a triazole group, inhibited cell adhesion, migration and invasion of highly metastatic B16F10 melanoma cells. TAP7f also exhibited antiangiogenic properties and reduced the expression levels of ß-catenin and metalloproteinases (MMPs) -2 and -9. To further explore TAP7f mechanism of action, we investigated by RT-qPCR whether the decrease in protein levels was caused by the inhibition of mRNA expression. Results showed a reduction in mRNA levels of both MMP-2 (58 ± 18 %) and -9 (46 ± 6 %), after 18 h of treatment with a 10 μM concentration of TAP7f. Under the same experimental conditions, TAP7f significantly reduced the mRNA levels of ß-catenin downstream targets cyclin-D1 (36 ± 9 %) and c-Myc (63 ± 18 %). Since increased amounts of integrin alfaV beta3 have been related to a higher metastatic potential of melanoma cells, we next studied the effect of TAP7f on integrin alfaV beta3 expression levels. RT-qPCR assays showed that TAP7f (10 μM) downregulated the mRNA levels of integrin alfaV (42 ± 8.3 %) and beta3 (37 ± 14 %). By flowcytometry, we demonstrated that TAP7f induced a decrease in the membrane expression of both integrin subunits. Based on results obtained in vitro, we explored TAP7f effect in an experimental lung metastasis model. C57BL/6J mice injected intravenously with B16F10 cells pretreated with 10 μM of TAP7f exhibited a ?50% reduction of lung nodules (p