Induction of p53 Phosphorylation at Serine 20 by Resveratrol Is Required to Activate p53 Target Genes, Restoring Apoptosis in MCF-7 Cells Resistant to Cisplatin.

Resistance to cisplatin (CDDP) is a major cause of cancer treatment failure, including human breast cancer. The tumor suppressor protein p53 is a key factor in the induction of cell cycle arrest, DNA repair, and apoptosis in response to cellular stimuli. This protein is phosphorylated in serine 15 and serine 20 during DNA damage repair or in serine 46 to induce apoptosis. Resveratrol (Resv) is a natural compound representing a promising chemosensitizer for cancer treatment that has been shown to sensitize tumor cells through upregulation and phosphorylation of p53 and inhibition of RAD51. We developed a CDDP-resistant MCF-7 cell line variant (MCF-7R) to investigate the effect of Resv in vitro in combination with CDDP over the role of p53 in overcoming CDDP resistance in MCF-7R cells. We have shown that Resv induces sensitivity to CDDP in MCF-7 and MCF-7R cells and that the downregulation of p53 protein expression and inhibition of p53 protein activity enhances resistance to CDDP in both cell lines. On the other hand, we found that Resv induces serine 20 (S20) phosphorylation in chemoresistant cells to activate p53 target genes such as PUMA and BAX, restoring apoptosis. It also changed the ratio between BCL-2 and BAX, where BCL-2 protein expression was decreased and at the same time BAX protein was increased. Interestingly, Resv attenuates CDDP-induced p53 phosphorylation in serine 15 (S15) and serine 46 (S46) probably through dephosphorylation and deactivation of ATM. It also activates different kinases, such as CK1, CHK2, and AMPK to induce phosphorylation of p53 in S20, suggesting a novel mechanism of p53 activation and chemosensitization to CDDP.

Histoenzymatic and morphometric analysis of muscle fiber type transformation during the postnatal development of the chronically food-deprived rat.

We analyze the effect of chronic undernourishment on extensor digitorum longus (EDL) muscle maturation in the rat. Cytochrome c oxidase (COX) and alkaline ATPase histoenzymatic techniques were used to determine the relative proportion of different fiber types (oxidative/glycolytic and type I, IIa/IId, or IIb, respectively) and their cross-sectional area in control and undernourished EDL muscles at several postnatal (PN) ages. From PN days 15 to 45, undernourished EDL muscles showed predominance of oxidative and type IIa/IId fibers, but from PN days 60 to 90, there were a larger proportion of oxidative fibers and an equal proportion of type IIa/IId and IIb fibers. Meanwhile, in adult stages (from PN days 130-365), the relative proportion of fiber types in control and undernourished EDL muscles showed no significant differences. In addition, from PN days 15 to 90, there was a significant reduction in the cross-sectional area of all fibers (slow: 13-53%; intermediate: 24-74%; fast: 9-80%) but no differences from PN days 130 to 365. It is suggested that chronic undernourishment affects the maturation of fast-type muscle fibers only at juvenile stages (from PN days 15-45) and the probable occurrence of adaptive mechanisms in muscle fibers, allowing adult rats to counterbalance the alterations provoked by chronic food deprivation.