Resveratrol-induced apoptosis in MCF-7 human breast cancer cells involves a caspase-independent mechanism with downregulation of Bcl-2 and NF-κB

Resveratrol (RES), a chemopreventive molecule, inhibits the proliferation of tumor cells of different etiologies. We previously showed that RES alters the cell cycle and induces apoptosis in MCF-7 breast tumor cells by interfering with the estrogen receptor (ER alpha)-dependent phosphoinositide 3-kinase (PI3K) pathway. Here, we analyzed signaling downstream of PI3K, to understand the mechanisms of RES-induced apoptosis. Apoptotic death by RES in MCF-7 was mediated by Bc1-2 downregulation since overexpression of this protein abolished apoptosis. Decreased Bc1-2 levels were not related to cytochrome c release, activation of caspases 3/8 or poly(ADP-ribose) polymerase proteolysis. However, RES decreased mitochondrial membrane potential and increased reactive oxygen species and nitric oxide production. NF-kappa B, a regulator of Bc1-2 expression, and calpain protease activity, a regulator of NF-kappa B, were both inhibited by RES. The patterns for NF-kappa B and calpain activities followed that of PI3K and were inhibited by LY294002. NF-kappa B inhibition coincided with diminished MMP9 activity and cell migration. These data suggest that RES-induced apoptosis in MCF-7 could involve an oxidative, caspase-independent mechanism, whereby inhibition of PI3K signaling converges to Bc1-2 through NF-kappa B and calpain protease activity. Therefore, Bc1-2 and NF-kappa B could be considered potential targets for the chemopreventive activity of RES in estrogen-responsive tumor cells. (c) 2005 Wiley-Liss. Inc.