Possible involvement of oxidative stress in trichloroethylene-induced genotoxicity in human HepG2 cells.
Trichloroethylene (TCE) is an environmental and industrial pollutant whose hepatotoxicity has been demonstrated in experimental animals. However, the mechanisms of the effects, in particular those related to its genotoxicity in humans, are not well understood. The aim of this study was to assess the genotoxic effects of TCE and to identify and clarify the mechanisms, using human hepatoma HepG2 cells. Exposure of the cells to TCE caused significant increase of DNA migration in comet assay and of micronuclei (MN) frequencies at all tested concentrations (0.5-4mM), respectively, which suggests that TCE caused DNA strand breaks and chromosome damage. The involvement of lipid peroxidation in the genotoxic properties of TCE was confirmed by using immunoperoxidase staining for 8-hydroxydeoxyguanosine (8-OHdG) and by measuring levels of thiobarbituric acid-reactive substances (TBARS). To elucidate the role of glutathione (GSH) in these effects, the intracellular GSH level was modulated by pre-treatment with buthionine-(S,R)-sulfoximine (BSO), a specific GSH synthesis inhibitor, and by co-treatment with N-acetylcysteine (NAC), a GSH precursor. It was found that depletion of GSH in HepG2 cells with BSO dramatically increased the susceptibility of HepG2 cells to TCE-induced cytotoxicity and DNA damage, while when the intracellular GSH content was elevated by NAC, the DNA damage induced by TCE was almost completely prevented. These results indicate that TCE exerts genotoxic effects in HepG2 cells, probably through DNA damage by oxidative stress; GSH, as a main intracellular antioxidant, is responsible for cellular defense against TCE-induced DNA damage.
Hu
Cunli
C
Department of Toxicology, Dalian Medical University, 465 Zhongshan Road, Dalian, 116027 Liaoning, China.
Jiang
Liping
L
Geng
Chengyan
C
Zhang
Xiaomei
X
Cao
Jun
J
Zhong
Laifu
L
D000080242:8-Hydroxy-2'-Deoxyguanosine; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D005978:Glutathione / Q000494:pharmacology; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology*; D048629:Micronuclei, Chromosome-Defective / Q000139:chemically induced; D009152:Mutagenicity Tests; D018384:Oxidative Stress / Q000502:physiology*; D012997:Solvents / Q000633:toxicity; D014241:Trichloroethylene / Q000633:toxicity*; D014407:Tumor Cells, Cultured
D012997:Solvents; D014241:Trichloroethylene; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine; D005978:Glutathione