Document Detail


Hydrogen peroxide-induced apoptosis in HL-60 cells requires caspase-3 activation.
MedLine Citation:
PMID:  10193575     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Apoptosis has been associated with oxidative stress in biological systems. Caspases have been considered to play a pivotal role in the execution phase of apoptosis. However, which caspases function as executioners in reactive oxygen species (ROS)-induced apoptosis is not known. The present study was performed to identify the major caspases acting in ROS-induced apoptosis. Treatment of HL-60 cells with 50 microM hydrogen peroxide (H2O2) for 4 h induced the morphological changes such as condensed and/or fragmented nuclei, increase in caspase-3 subfamily protease activities, reduction of the procaspase-3 and a DNA fragmentation. To determine the role of caspases in H2O2-induced apoptosis, caspase inhibitors, acetyl-Tyr-Val-Ala-Asp-chloromethyl ketone (Ac-YVAD-cmk), acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO) and acetyl-Val-Glu-Ile-Asp-aldehyde (Ac-VEID-CHO), selective for caspase-1 subfamily, caspase-3 subfamily and caspase-6, respectively, were loaded into the cells using an osmotic lysis of pinosomes method. Of these caspase inhibitors, only Ac-DEVD-CHO completely blocked morphological changes, caspase-3 subfamily protease activation and DNA ladder formation in H2O2-treated HL-60 cells. This inhibitory effect was dose-dependent. These results suggest that caspase-3, but not caspase-1 is required for commitment to ROS-triggered apoptosis.
Authors:
T Matsura; M Kai; Y Fujii; H Ito; K Yamada
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Free radical research     Volume:  30     ISSN:  1071-5762     ISO Abbreviation:  Free Radic. Res.     Publication Date:  1999 Jan 
Date Detail:
Created Date:  1999-06-23     Completed Date:  1999-06-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9423872     Medline TA:  Free Radic Res     Country:  SWITZERLAND    
Other Details:
Languages:  eng     Pagination:  73-83     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Faculty of Medicine, Tottori University, Yonago, Japan. tmatsura@grape.med.tottori-u.ac.jp
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Chloromethyl Ketones / administration & dosage,  pharmacology
Apoptosis / drug effects*
Blotting, Western
Caspase 3
Caspases / antagonists & inhibitors,  metabolism*
Cell Nucleus / drug effects,  metabolism
DNA Fragmentation / drug effects
Dose-Response Relationship, Drug
Enzyme Activation / drug effects
HL-60 Cells
Humans
Hydrogen Peroxide / pharmacology*
Oligopeptides / administration & dosage,  pharmacology
Reactive Oxygen Species / metabolism
Time Factors
Chemical
Reg. No./Substance:
0/Amino Acid Chloromethyl Ketones; 0/N-acetyl-tyrosyl-valyl-alanyl-aspartyl chloromethyl ketone; 0/Oligopeptides; 0/Reactive Oxygen Species; 0/acetyl-aspartyl-glutamyl-valyl-aspartal; 7722-84-1/Hydrogen Peroxide; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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