Antitumor properties of two copper based compounds against 2D and 3D human colorectal cancer cell models.

RUIZ M.C; DI VIRGILIO A.L; BOLLATI-FOGOLIN M; PERELMULTER K; LEON I.E

Congreso; Reunión Científica Anual de Sociedades de Biociencia (SAIC-SAFE-SAP-SAB); 2019

Many functions of metal ion have stimulated the development of new metallodrugs. The synthesis of new copper complexes is potentially attractive as anticancer agents; whose properties are determined by the nature of ligands bound to the metal ion. This study evaluates the action of two cooper complexes, CuII(dmp)2(CH3CN)](ClO4)2 (1) and CuII(phen)2](ClO4) (2), against human colorectal cancer cells. An in vitro cytotoxicity assay was carried out on cultured HT29, Caco-2 and LS174T cell lines monolayer. To get insight over the mechanism of action, we study the role of ROS generation, using DHR123 probe and the mitochondrial membrane potential (MMP) with DiOC6. The migration process was investigated with gelatin zymography as well. Moreover, apoptosis was studied with anexinV/IP and caspase 3 assays by flow citometry, adding studies of morphological changes with fluorescent microscopy. Furthermore, the cytotoxicity was studied with IP on 3D cell model derived from HT29 cells. In addition, NF-κB pathway suppression was investigated. Both complexes caused significant cytotoxicity in all cell lines, proving that complex 1 is more active (IC50 values for HT29 are 1.45 vs 2.76 µM, for Caco-2 2.32 vs 6.48 µM, for LS174T 1.44 vs 2.54 µM for 1 and 2, respectively). It can be noticed that complex 1 increased ROS in all cell lines and the MMP decreased with the 24 h treatment. Flow cytometric analysis revealed that theses complexes induce apoptotic in a dose and time dependent manner. These results are validated used microscopy. Complex 1 also attenuated the secretion of the metalloproteinases 2 and 9. On cell spheroids the IC50 values were 18.32 µM for 1 and 19.12 µM for 2. Interestingly, both complexes decrease the NF-κB expression in cell monolayer and spheroids, showing an inhibition of this pathway. In conclusion, both compounds display an antitumor activity; however 1 was more effective in monolayer and 3D model than 2, being a candidate for in vivo experiments.