| Contribution of death receptor and mitochondrial pathways to Fas-mediated apoptosis in the prostatic carcinoma cell line PC3. | |
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MedLine Citation:
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PMID: 11987151 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: Two main pathways of apoptosis in mammalian cells have been described: the death receptor pathway and the mitochondrial pathway. Two different cell types have been identified for Fas-mediated apoptosis, each using almost exclusively one of two different signaling pathways. Human prostatic carcinoma cell line, PC3 is sensitive to Fas-mediated apoptosis, but relation of receptor and mitochondrial pathways is not clear. METHODS: Cell viability was estimated by calcein assay. Apoptosis was determined by preparation of DNA ladder. Expression of Fas-associated death domain-dominant negative (FADD-DN) and Bcl-2, activation of caspases, PARP, DFF45, Bid cleavage, and cytochrome c release were assessed using Western blotting techniques. [(35)S] Methionine-labeled caspase-3 was transcribed in vitro and translated using the TNT kit (Promega). A vector containing caspase-3 was prepared by the ligation of EcoR I/BamHI flanked PCR fragment of full size caspase-3 cDNA into pBlusckript II SK(+/-) (Stratagen). RESULTS: Overexpression of both FADD-DN and Bcl-2 genes prevent Fas-mediated apoptosis in PC3. As predicted, overexpression of FADD-DN prevented activation of caspase-8 and Bid cleavage and attenuated the release of cytochrome c and activation of caspases -2, -7, and -9. Bcl-2 overexpression did not affect caspase-8 activation and cleavage of Bid but blocked the release of cytochrome c and activation of mitochondria localized caspases -2, -7, and-9. Overexpression of FADD-DN and Bcl-2 affected the activation of caspase-3 and PARP cleavage differently: FADD-DN attenuated the activation of caspase-3 and PARP cleavage whereas Bcl-2 overexpression prevented caspase-3 activation and completely blocked cleavage of PARP. CONCLUSIONS: These data suggest that activation of caspase-8 is necessary but not sufficient to complete Fas-mediated apoptosis in PC3 cells without activation of the mitochondrial pathway. In addition, caspase-3 activation after Fas-receptor ligation involves two steps and is dependent on mitochondrial activation. |
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Authors:
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Natalya V Guseva; Agshin F Taghiyev; Oskar W Rokhlin; Michael B Cohen |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: The Prostate Volume: 51 ISSN: 0270-4137 ISO Abbreviation: Prostate Publication Date: 2002 Jun |
Date Detail:
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Created Date: 2002-05-23 Completed Date: 2002-06-19 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8101368 Medline TA: Prostate Country: United States |
Other Details:
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Languages: eng Pagination: 231-40 Citation Subset: IM |
Copyright Information:
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Copyright 2002 Wiley-Liss, Inc. |
Affiliation:
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Department of Pathology, The University of Iowa, Iowa City, Iowa 52242-1087, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Antigens, CD95
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physiology* Apoptosis* Blotting, Western Caspase 8 Caspase 9 Caspases / pharmacology Gene Expression Regulation, Neoplastic* Humans Male Mitochondria / physiology* Polymerase Chain Reaction Prostatic Neoplasms / pathology* Proto-Oncogene Proteins c-bcl-2 / biosynthesis, pharmacology* |
| Chemical | |
Reg. No./Substance:
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0/Antigens, CD95; 0/Proto-Oncogene Proteins c-bcl-2; EC 3.4.22.-/CASP8 protein, human; EC 3.4.22.-/CASP9 protein, human; EC 3.4.22.-/Caspase 8; EC 3.4.22.-/Caspase 9; EC 3.4.22.-/Caspases |
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