The effect of alpha-phenyl-N-t-butylnitrone on ionizing radiation-induced apoptosis in U937 cells.
Ionizing radiation induces the production of reactive oxygen species (ROS), which play an important causative role in apoptotic cell death. alpha-Phenyl-N-t-butylnitrone (PBN) is one of the most widely used spin-trapping compounds for investigating the existence of free radicals in biological systems. We investigated the effects of PBN on ionizing radiation-induced apoptosis in U937 cells. Upon exposure to 2 Gy of gamma-irradiation, there was a distinct difference between the control cells and the cells pre-treated with 2 mM PBN for 2 h in regard to apoptotic parameters, cellular redox status, mitochondria function and oxidative damage to cells. PBN effectively suppressed morphological evidence of apoptosis and DNA fragmentation in U937 cells exposed to ionizing radiation. The [GSSG]/[GSH+GSSG] ratio and the generation of intracellular ROS were higher and the [NADPH]/[NADP+ +NADPH] ratio was lower in control cells compared to PBN-treated cells. The ionizing radiation-induced mitochondrial damage reflected by the altered mitochondrial permeability transition, the increase in the accumulation of ROS, and the reduction of ATP production were significantly higher in control cells compared to PBN-treated cells. PBN pre-treated cells showed significant inhibition of apoptotic features such as activation of caspase-3, up-regulation of Bax and p53, and down-regulation of Bcl-2 compared to control cells upon exposure to ionizing radiation. This study indicates that PBN may play an important role in regulating the apoptosis induced by ionizing radiation presumably through scavenging of ROS.
Lee
Jin Hyup
JH
School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Taegu, 702-701, South Korea.
Park
Jeen-Woo
JW
D000255:Adenosine Triphosphate / Q000096:biosynthesis; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D003497:Cyclic N-Oxides; D053938:DNA Fragmentation / Q000187:drug effects; D005720:Gamma Rays; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008928:Mitochondria / Q000502:physiology; D009249:NADP / Q000378:metabolism; D009589:Nitrogen Oxides / Q000494:pharmacology*; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D019097:Spin Trapping; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D020298:U937 Cells; D051028:bcl-2-Associated X Protein / Q000378:metabolism
C494826:BAX protein, human; D003497:Cyclic N-Oxides; D009589:Nitrogen Oxides; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C029217:phenyl-N-tert-butylnitrone; D009249:NADP; D000255:Adenosine Triphosphate; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D005978:Glutathione