The role of oxidative stress in acrolein-induced DNA damage in HepG2 cells.
This study evaluated the role of oxidative stress in acrolein-induced DNA damage, using HepG2 cells. Using the standard single cell gel electrophoresis (SCGE) assay, a significant dose-dependent increment in DNA migration was detected at lower concentrations of acrolein; but at the higher tested concentrations, a reduction in the migration was observed. Post-incubation with proteinase K significantly increased DNA migration in cells exposed to higher concentrations of acrolein. These results indicated that acrolein caused DNA strand breaks and DNA-protein crosslinks (DPC). To elucidate the oxidatively generated DNA damage mechanism, the 2,7-dichlorofluorescein diacetate (DCFH-DA) and o-phthalaldehyde (OPT) were used to monitor the levels of reactive oxygen species (ROS) and glutathione (GSH), respectively. The present study showed that acrolein induced the increased levels of ROS and depletion of GSH in HepG2 cells. Moreover, acrolein significantly caused 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) formation in HepG2 cells. These results demonstrate that the DNA damage induced by acrolein in HepG2 cells is related to the oxidative stress.
Li
Longjie
L
Department of Toxicology, Dalian Medical University, No. 9, West Segment of South lushun Road, Lushunkou District, Dalian 116044, Liaoning, PR China.
Jiang
Liping
L
Geng
Chengyan
C
Cao
Jun
J
Zhong
Laifu
L
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000171:Acrolein / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival; D020552:Comet Assay; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D004586:Electrophoresis; D019286:Endopeptidase K / Q000494:pharmacology; D005452:Fluoresceins / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D018384:Oxidative Stress; D017382:Reactive Oxygen Species; D009764:o-Phthalaldehyde / Q000494:pharmacology
D003432:Cross-Linking Reagents; D005452:Fluoresceins; D017382:Reactive Oxygen Species; C029569:diacetyldichlorofluorescein; D009764:o-Phthalaldehyde; D000171:Acrolein; D000080242:8-Hydroxy-2'-Deoxyguanosine; D019286:Endopeptidase K; D003849:Deoxyguanosine; D005978:Glutathione