In summary, based on gene expression profile analysis re sults, we can speculate that different molecular mecha nisms may contribute to the anticancer effect of D6 in melanoma cells i) the induction of a cell stress MEK162 IC50 response that triggers the ER stress mediated apoptosis pathway. ii) the up regulation of p53 signalling, which promotes p21 and GADD45 dependent cell cycle arrest as well as mito chondrial apoptosis based on Inhibitors,Modulators,Libraries Noxa over expression. iii) the down modulation of several growth signals, like both PI3K and NF kB pathways, and c kit receptor. The diagram in Inhibitors,Modulators,Libraries Figure 5 summarizes the major gene expression changes in duced by D6 in melanoma cells and hypothesizes the pos sible intervention of these changes in depicting cellular fate.

Conclusions Altogether, our findings contribute to unveil the molecular mechanisms underlying the anti tumour activity of D6 in melanoma cells. Based on such results, we can speculate that a) p53 protein Inhibitors,Modulators,Libraries may play a key role in sustaining the anticancer effects exerted by D6 on melanoma cells. b) in duction of strong cell stress responses may contribute to the reinforcement of the proapoptotic trend of p53 sig nalling. and c) down modulation of several growth signals, as well as the under expression of cell cycle regulators might be involved in cell growth inhibition. This last aspect seems to be peculiar of the response to D6 treatment in melanoma cells, being absent in D6 treated fibroblasts expression profile. Although our analyses were not exhaustive, data here presented strongly indicate that a huge amount of molecu lar changes does participate in determining the molecular mechanism of action of D6 on melanoma cells.

Gene ex pression profile analyses on additional melanoma cell Inhibitors,Modulators,Libraries lines are currently in progress, in order to either confirm our findings in a larger samples collection or evaluate the effects of D6 on both primary and metastatic tumour derived cell lines. Methods Cell cultures and D6 treatments Malignant melanoma LB24Dagi cell line was obtained from Inhibitors,Modulators,Libraries the Department of Molecular and Trichostatin A TSA Cellular Biology at the Istituto Dermopatico dellImmacolata in Rome. Normal human fibroblast BJ were purchased from the American Type Culture Collec tion. All cells were grown in RPMI media, supplemented with 10% FBS and penicillin streptomycin, as described. The B unsaturated ketone D6 has been synthesized in our lab as previously described. Stock solution of D6 was prepared by dis solving D6 in DMSO to a final concentration of 100 mM and stored at ?20 C. Working solutions of D6 were prepared daily as previously described.