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

ELIZABETH BARRIONUEVO; FLORENCIA CAYROL; GRACIELA A. CREMASCHI; PATRICIA G. CORNIER; CARINA M. L. DELPICCOLO; DORA B. BOGGIÁN,; ERNESTO G. MATA; LEONOR P. ROGUIN; VIVIANA BLANK

Congreso; Reunión Conjunta SAIC, SAFE, SAB, SAF; 2019

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 β-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 β-catenin downstream targets cyclin-D1 (38±5%) and c-Myc (65±12%). Since increased amounts of integrin αVβ3 have been related to a higher metastatic potential of melanoma cells, we next studied the effect of TAP7f on integrin αVβ3 expression levels. RT-qPCR assays showed that TAP7f (10 μM) downregulated the mRNA levels of integrin αV (42±8.3%) and β3 (37±14%). By flow cytometry, 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