Effetti del Cadmio e del 5 Fluorouracile su cellule di tumore mammario MCF7: analisi morfostrutturale, di espressione genica e proteica

Cadmium (Cd) is considerate one of the most dangerous metals, for men’s health. 5-Fluorouracil (5-Fu) is an antimetabolite used in chemotherapy protocols for solid tumors. The MCF7 human breast cancer cell had been used as cellular model. We analysed the correlation between MCF-7 ultrastructural alterations, Actin and Tubulin cytoskeleton proteins and gene and protein expressions.MCF-7 were treated with Cd(5μM, 20μM, 40μM) and 5-FU(1.5μM), for 24h and 48h, for TEM, SEM and Immunofluorescence. For gene and protein expression MCF-7 were treated with the Ic 50 (Cd 3μM, and 5-FU 1.5μM) for 6h, 24h, 48h. Gene expression ofBcl2,P53,Bax,Caspase8,Caspase 9,C-Myc,Cyclin A1,Cyclin D1, was studied with qRT-PCR, proteins were analyzed with Western Blot.In cells Cd treated we observed an intense metabolic activity with alterated mitochondria and alterations in Actin distribution while Tubulin branched out in the cytoplasm. 5-FU-treated cells showed cytotoxicity signs with the presence of pore-like in the membrane cell and degeneration of cytoplasm and nuclei. The addition of 5-FU to the cells Cd treated did not induce significant changes in comparison with the cells Cd treated.Genes and proteins expressions were significant at 24h. 5-FU+Cd treated cells, had an increase of anti-apoptotic expression ofBcl-2,CyclinsandC-Myc.Our results suggest that Cd has the ability to stop the cytotoxic effects of 5-FU, used in our experiments and in traditional breast cancer therapy.