GEA 3162, a peroxynitrite donor, induces Bcl-2-sensitive, p53-independent apoptosis in murine bone marrow cells

• Published in: Biochemical pharmacology Vol. 74; no. 7; pp. 1039 - 1049
• Main Authors: Taylor, Emma L., Li, John T., Tupper, Joan C., Rossi, Adriano G., Winn, Robert K., Harlan, John M.
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.10.2007; Elsevier Science
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Bcl-2; Biological and medical sciences; Bone Marrow Cells - cytology; Bone Marrow Cells - drug effects; Bone Marrow Cells - metabolism; Caspase; Caspases - metabolism; Cell Line; Cell Survival; Dose-Response Relationship, Drug; Enzyme Activation; Medical sciences; Mice; Mitochondria - drug effects; Mitochondria - metabolism; Mitogen-Activated Protein Kinases - metabolism; Myeloid; Nitric Oxide Donors - pharmacology; p53; Peroxynitrite; Pharmacology. Drug treatments; Proto-Oncogene Proteins c-bcl-2 - metabolism; Triazoles - pharmacology; Tumor Suppressor Protein p53 - metabolism; Bcl-2; NO; Peroxynitrite; JNK; Jaws II-GFP; Caspase; MAP kinase; Myeloid; p53; GEA 3162; GFP; Jaws II-Bcl-2; STSP; ONOO; Apoptosis; Enzyme; Cysteine endopeptidases; p53 Protein; Rodentia; In vitro; Peptidases; Vertebrata; Mammalia; TP53 Gene; Donor; Mouse; Cell death; Animal; C-Onc gene; Bone marrow; Hydrolases; Protooncogene; Tumor suppressor gene
• ISSN: 0006-2952; 1873-2968
• DOI: 10.1016/j.bcp.2007.06.028

Apoptosis may be regulated by oxidants such as peroxynitrite (ONOO−). The tumour suppressor, p53, has been reported to play a crucial role in apoptosis induced by oxidants, therefore we assessed the ability of a ONOO− donor, GEA 3162, to activate caspases and induce mitochondrial permeability in a p53-deficient murine bone marrow cell line, Jaws II. Furthermore, these cells were stably transfected with Bcl-2, in order to investigate the impact of this survival protein on ONOO−-induced apoptosis. GEA 3162 activated caspases and induced loss of mitochondrial membrane potential in Jaws II cells. In particular, caspases 3 and 2 were activated, alongside minor activation of caspases 8 and 9, and apoptosis was partially dependent upon p38 MAP kinase activation, with little or no role for JNK. Overexpression of Bcl-2 abolished activation of all caspases and reduced the change in mitochondrial membrane potential. Thus, we have demonstrated that the ONOO− donor, GEA 3162, induces apoptosis in Jaws II murine myeloid cells despite lacking functional p53, via a pathway that principally involves caspases 2 and 3 and mitochondrial changes. This is blocked by overexpression of Bcl-2 via a mechanism that does not appear to merely reflect stabilisation of the mitochondrial membrane.