Cupric nitrilotriacetate-induced apoptosis in HL-60 cells association with lipid peroxidation, release of cytochrome C from mitochondria, and activation of caspase-3.
Oxidative stress may be a common mechanism underlying various forms of cell death, including necrosis and apoptosis. The authors have reported previously that the cupric nitrilotriacetate (Cu-NTA), a renal carcinogen, induces oxidative DNA damage and apoptosis in HL-60 human leukemia cells (Ma, Y., et al. Free Radic. Biol Med. 25:568-575; 1998). The focus of this investigation was to examine the possible pathway of the apoptosis induced by Cu-NTA. Results of the present study demonstrated that after exposure of HL-60 cells to Cu-NTA, an increase in lipid hydroperoxide and loss of mitochondrial membrane potential (deltaphim) were observed, followed by the increase in cytosolic cytochrome c that was released from the mitochondria. These events proceeded and triggered the activation of caspase-3 (CPP32/apopain/Yama), resulting in the degradation of poly (ADP-ribose) polymerase and DNA fragmentation. The antioxidants, N-acetylcysteine and glutathione, protected the loss of deltaphim and blocked the apoptosis induced by Cu-NTA. In addition, Ac-DEVD-CHO, a specific inhibitor of caspase-3, inhibited Cu-NTA-induced apoptosis. These results suggested that Cu-NTA-induced apoptosis in HL-60 cells was, at least in part, triggered by free radical-induced lipid peroxidation of membrane, which induced the release of cytochrome c from mitochondria and activation of caspase-3.
Ma
Y
Y
Department of Pathology, Okayama University Medical School, Shikata-cho, Japan.
Ogino
T
T
Kawabata
T
T
Li
J
J
Eguchi
K
K
Okada
S
S
D000111:Acetylcysteine / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D003300:Copper / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004789:Enzyme Activation; D005978:Glutathione / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D015227:Lipid Peroxidation; D008928:Mitochondria / Q000378:metabolism*; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D009942:Organometallic Compounds / Q000378:metabolism / Q000494:pharmacology*
D000975:Antioxidants; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D003574:Cytochrome c Group; D009842:Oligopeptides; D009942:Organometallic Compounds; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; D003300:Copper; C060581:cupric nitrilotriacetate; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D005978:Glutathione; D009571:Nitrilotriacetic Acid; D000111:Acetylcysteine; D004121:Dimethyl Sulfoxide