Contribution of death receptor and mitochondrial pathways to Fas-mediated apoptosis in the prostatic carcinoma cell line PC3.

摘要: BACKGROUND Two main pathways of apoptosis in mammalian cells have been described: the death receptor pathway and mitochondrial pathway. Two different cell types identified for Fas-mediated apoptosis, each using almost exclusively one two signaling pathways. Human prostatic carcinoma line, PC3 is sensitive to but relation not clear. METHODS Cell viability was estimated by calcein assay. Apoptosis determined preparation DNA ladder. Expression Fas-associated domain-dominant negative (FADD-DN) Bcl-2, activation caspases, PARP, DFF45, Bid cleavage, cytochrome c release were assessed Western blotting techniques. [35S] Methionine-labeled caspase-3 transcribed vitro translated TNT kit (Promega). A vector containing prepared ligation EcoR I/BamHI flanked PCR fragment full size cDNA into pBlusckript II SK(+/−) (Stratagen). RESULTS Overexpression both FADD-DN Bcl-2 genes prevent PC3. As predicted, overexpression prevented caspase-8 cleavage attenuated caspases -2, -7, -9. did affect blocked mitochondria localized and–9. Overexpression affected PARP differently: whereas completely PARP. CONCLUSIONS These data suggest that necessary sufficient complete without In addition, after Fas-receptor involves steps dependent on activation. Prostate 51: 231–240, 2002. © 2002 Wiley-Liss, Inc.