Mercury-induced apoptosis in human lymphocytes: caspase activation is linked to redox status.
There is growing evidence that heavy metals, in general, and mercurial compounds, in particular, are toxic to the human immune system. We have previously shown that methyl mercuric chloride (MeHgCl) is a potent human T-cell apoptogen; moreover, mitochondria appear to be a target organelle for the induction of cell death. The objective of this study was to determine the impact of MeHgCl on mitochondrial function in lymphocytes in terms of modulating reactive oxygen species (ROS) generation, thiol status, and caspase activation. Using the fluorescent probe, 3,3'-dihexyloxacarbocyanine, we demonstrated that exposure to MeHgCl for 1 h resulted in a profound decrease in the mitochondrial transmembrane potential. We next observed the release of cytochrome c from mitochondria into the cytosol; significant translocation was noted between 4 and 8 h following treatment with mercury. ROS generation was monitored by following the conversion of dihydroethidium to the fluorescent product, ethidium. Kinetic analysis indicated that ROS generation was maximal after 16 h of exposure to MeHgCl. The toxicant also depleted the thiol reserves of the cell; glutathione levels were depleted in a dose-dependent fashion reaching minimal levels at 16 h. Real-time RT-PCR analysis demonstrated a significant reduction in both glutathione S-transferase and glutathione peroxidase gene expression in mercury-treated cells. Finally, after 16 h of treatment with MeHgCl, we observed activation of caspase-8, -9, and -3 along with increased expression of caspase-8 and -9. We propose that the target organelle for MeHgCl is the mitochondrion and that induction of oxidative stress is critical to activation of death-signaling pathways. Additonally, mercury acts as a genotoxin significantly altering the expression of genes that affect cell survival and apoptosis.
Shenker
Bruce J
BJ
Department of Pathology, University of Pennsylvania School of Dental Medicine, Philadelphia, PA 19104-6002, USA. shenker@pobox.upenn.edu
Pankoski
Lisa
L
Zekavat
Ali
A
Shapiro
Irving M
IM
D000328:Adult; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism; D004789:Enzyme Activation; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008564:Membrane Potentials / Q000502:physiology; D008767:Methylmercury Compounds / Q000494:pharmacology* / Q000633:toxicity; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D008954:Models, Biological; D010084:Oxidation-Reduction; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism
D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D003574:Cytochrome c Group; D005456:Fluorescent Dyes; D008767:Methylmercury Compounds; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D005982:Glutathione Transferase; D020169:Caspases; D005978:Glutathione