Wine consumption reduced postprandial platelet sensitivity against platelet activating factor in healthy men.

PURPOSE: Platelet activating factor (PAF) is a potent inflammatory and thrombotic mediator that participates in the initiation and prolongation of atherosclerosis. The aim of the present study was to evaluate the potential effect of wine consumption on platelet aggregation against PAF. METHODS: The study had cross-over design. Ten healthy men participated in four daily trials on separate days: They consumed a standardized meal along with white wine, Robola variety (trial R), or red wine, Cabernet Sauvignon variety (trial CS), or an ethanol solution (trial E), or water (trial W). Blood samples were collected before and after meal consumption and at several time points during the next 6 h. Platelet aggregation against PAF (EC50 values) and several blood biomarkers were measured, and incremental areas under the curve (iAUC) were calculated. RESULTS: A significant trial effect was found in platelet sensitivity against PAF (p trial = 0.01). Moreover, the iAUC-PAF EC50 of CS trial was higher compared to both iAUC-PAF EC50 of E and W trials (P = 0.04, P = 0.02). Plasminogen activator inhibitor-1 iAUC was higher in all alcoholic beverages compare with the one of W trial (P E  = 0.05, P R  = 0.01, P CS = 0.01). Triacylglycerol iAUC increased significantly only in E compared to W trial (P = 0.04) and were significantly lower at 60-120 min in wine trials compared to the one of E (P < 0.05). CONCLUSIONS: Wine consumption improved platelet sensitivity independently of alcohol, kept triacylglycerols at lower levels during their postprandial elevation, and did not affect plasminogen activator inhibitor-1 levels more adversely than ethanol per se.

Thymidylate synthase inhibition induces p53-dependent and p53-independent apoptotic responses in human urinary bladder cancer cells.

PURPOSE: In search for more effective clinical protocols, the antimetabolite drug 5-fluorouracil (5-FU) has been successfully included in new regimens of bladder cancer combination chemotherapy. In the present study, we have investigated the effects of 5-FU treatment on apoptosis induction in wild-type and mutant p53 urinary bladder cancer cells. METHODS: We have used MTT-based assays, FACS analysis, Western blotting and semi-quantitative RT-PCR in RT4 and RT112 (grade I, wild-type p53), as well as in T24 (grade III, mutant p53) and TCCSUP (grade IV, mutant p53) human urinary bladder cancer cell lines. RESULTS: In the urothelial bladder cancer cell lines RT4 and T24, 5-FU-induced TS inhibition proved to be associated with cell type-dependent (a) sensitivity to the drug, (b) Caspase-mediated apoptosis, (c) p53 stabilization and activation, as well as Rb phosphorylation and E2F1 expression and (d) transcriptional regulation of p53 target genes and their cognate proteins, while an E2F-dependent transcriptional network did not seem to be critically engaged in such type of responses. CONCLUSIONS: We have shown that in the wild-type p53 context of RT4 cells, 5-FU-triggered apoptosis was prominently efficient and mainly regulated by p53-dependent mechanisms, whereas the mutant p53 environment of T24 cells was able to provide notable levels of resistance to apoptosis, basically ascribed to E2F-independent, and still unidentified, pathways. Nevertheless, the differential vulnerability of RT4 and T24 cells to 5-FU administration could also be associated with cell-type-specific transcriptional expression patterns of certain genes critically involved in 5-FU metabolism.

Grade-dependent effects on cell cycle progression and apoptosis in response to doxorubicin in human bladder cancer cell lines.

Doxorubicin is an important component of combination therapy for muscle-invasive urinary bladder cancer. Treatment with this topoisomerase II poison is able to interfere with cell cycle progression and lead to cancer cell death. Using FACS analysis, Western immunoblotting and semi-quantitative RT-PCR, we studied the effects of doxorubicin on cell cycle progression and apoptosis, and also explored the possibility of using groups of genes as biomarkers of prognosis and/or response to doxorubicin treatment in human urinary bladder cancer cells. Doxorubicin induced dose-dependent G2/M and/or G1/S cell cycle arrest, followed by grade- and dose-dependent reduction in the amount of the cytosolic trimeric form of FasL, activation of Caspase-8, Caspase-9, Caspase-3, cleavage of PARP, Lamin A/C, Bcl-XL/S and interestingly Hsp90, and finally cell death. Data presented here also suggest the use of the expression patterns of Cyclin-E2, Cyclin-F, p63, p73, FasL, TRAIL, Tweak, Tweak-R, XAF-1, OPG and Bok genes for identification of the differentiation grade, and Cyclin-B2, GADD45A, p73, FasL, Bik, Bim, TRAIL, Fas, Tweak-R, XAF-1, Bcl-2, Survivin, OPG, DcR2 and Bcl-XL genes for the detection of response to doxorubicin in human bladder cancer cells.