Reactive oxygen species and mitochondria mediate the induction of apoptosis in human hepatoma HepG2 cells by the rodent peroxisome proliferator and hepatocarcinogen, perfluorooctanoic acid.
We have previously shown that one of the most potent rodent hepatocarcinogens, perfluorooctanoic acid (PFOA), induces apoptosis in human HepG2 cells in a dose- and time-dependent manner. In this study we have investigated the involvement of reactive oxygen species (ROS), mitochondria, and caspase-9 in PFOA-induced apoptosis. Treatment with 200 and 400 microM PFOA was found to cause a dramatic increase in the cellular content of superoxide anions and hydrogen peroxide after 3 h. Measurement of the mitochondrial transmembrane potential (Delta Psi(m)) after PFOA treatment showed a dissipation of Delta Psi(m) at 3 h. Caspase-9 activation was seen at 5 h after treatment with 200 microM PFOA. In order to evaluate the importance of these events in PFOA-induced apoptosis, cells were cotreated with PFOA and N-acetylcysteine (NAC), a precursor of glutathione, or Cyclosporin A (CsA), an inhibitor of mitochondrial permeability transition pore (MPT pore). NAC reduced Delta Psi(m) dissipation, caspase 9 activation, and apoptosis, indicating a role for PFOA-induced ROS. In addition, CsA also reduced Delta Psi(m) dissipation, caspase 9 activation, and apoptosis, indicating a role for PFOA-induced opening of the MPT pore. In summary, we have delineated a ROS and mitochondria-mediated pathway for induction of apoptosis by PFOA.
Panaretakis
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T
Unit of Biochemical Toxicology, Department of Biochemistry, Wallenberg Laboratory, Stockholm University, S-106 91 Stockholm, Sweden. theoharis.panaretakis@cck.ki.se
Shabalina
I G
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Grandér
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Shoshan
M C
MC
DePierre
J W
JW
D000111:Acetylcysteine / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002210:Caprylates / Q000494:pharmacology*; D002273:Carcinogens / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D005434:Flow Cytometry; D005466:Fluorocarbons / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D008113:Liver Neoplasms / Q000473:pathology*; D008564:Membrane Potentials; D008930:Mitochondria, Liver / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D014407:Tumor Cells, Cultured
D002210:Caprylates; D002273:Carcinogens; D005466:Fluorocarbons; D017382:Reactive Oxygen Species; D013481:Superoxides; D016572:Cyclosporine; C023036:perfluorooctanoic acid; D006861:Hydrogen Peroxide; C506433:CASP9 protein, human; D053453:Caspase 9; D020169:Caspases; D000111:Acetylcysteine