Document Detail


N-methyl-N'-nitro-N-nitrosoguanidine activates multiple cell death mechanisms in human fibroblasts.
MedLine Citation:
PMID:  17678437     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Response to genotoxic stress may trigger the activation of distinct mechanisms that serve to promote cell death, including apoptosis and necrosis. In this study we examined the response of human fibroblasts, either proficient or deficient for the damage-activated protein kinase ataxia telangiectasia-mutated (ATM), to the alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Analysis of both long- and short-term viability shows that both ATM-proficient YZ-5 and ATM-deficient EBS-7 fibroblasts display a cytotoxic response to MNNG. Consistent with activation of apoptosis in response to MNNG, we observed increased caspase-3 cleavage and activity, appearance of fragmented nuclei, and increased staining with annexin V in both ATM-proficient and -deficient fibroblasts. Flow cytometry demonstrated that these cell lines also display a nonapoptotic cell death in response to MNNG. This form of cell death is associated with activation of poly-ADP ribose polymerase (PARP), and analysis of PARP activity indicated increased protein poly(ADP-ribosylation) in YZ-5 when compared to EBS-7. This PARP activity was accompanied by apoptosis-inducing factor release and translocation from the mitochondria to the nucleus. Finally, the PARP inhibitor 3,4-dihydro-5-[4-(1-piperidinyl)butoxy]-1(2H)-isoquinolinone (DPQ) or the caspase-3 inhibitor benzyloxycarbonyl-VAD-fluoromethyl ketone dramatically diminished the cytotoxic response to MNNG, reinforcing the roles for apoptotic and nonapoptotic cell death in human fibroblasts treated with MNNG. From these findings, we conclude that MNNG induces a heterogeneous death response in human fibroblasts.
Authors:
Michael W Lee; Wan-Ju Kim; Dillon I Beardsley; Kevin D Brown
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  DNA and cell biology     Volume:  26     ISSN:  1044-5498     ISO Abbreviation:  DNA Cell Biol.     Publication Date:  2007 Sep 
Date Detail:
Created Date:  2007-09-21     Completed Date:  2007-10-23     Revised Date:  2011-11-02    
Medline Journal Info:
Nlm Unique ID:  9004522     Medline TA:  DNA Cell Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  683-94     Citation Subset:  IM    
Affiliation:
Department of Biochemistry and Molecular Biology and the UF-Shands Cancer Center Program in Cancer Genetics, Epigenetics and Tumor Virology, University of Florida College of Medicine, Gainesville, Florida 32610, USA.
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MeSH Terms
Descriptor/Qualifier:
Annexin A5 / metabolism
Caspases / metabolism
Cell Cycle Proteins / genetics,  physiology*
Cell Death / drug effects
Cell Nucleus / metabolism
Cells, Cultured
Chromatin / metabolism
Colony-Forming Units Assay
DNA-Binding Proteins / genetics,  physiology*
Enzyme Inhibitors / pharmacology
Fibroblasts / drug effects*,  metabolism
Humans
Immunoblotting
Methylnitronitrosoguanidine / pharmacology*
Poly(ADP-ribose) Polymerases / metabolism
Protein Transport
Protein-Serine-Threonine Kinases / genetics,  physiology*
Tumor Suppressor Proteins / genetics,  physiology*
Grant Support
ID/Acronym/Agency:
R01-CA102289/CA/NCI NIH HHS; T32-CA09126/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Annexin A5; 0/Cell Cycle Proteins; 0/Chromatin; 0/DNA-Binding Proteins; 0/Enzyme Inhibitors; 0/Tumor Suppressor Proteins; 70-25-7/Methylnitronitrosoguanidine; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 2.7.11.1/Protein-Serine-Threonine Kinases; EC 2.7.11.1/ataxia telangiectasia mutated protein; EC 3.4.22.-/Caspases

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