[Mercuric chloride induces apoptosis in MDCK cells]

Mercury is a hazardous organ-specific environmental contaminant. It exists in a wide variety of physical and chemical states, each of which has unique characteristics for the target organ specificity. Exposure to mercury vapor and to organic mercury compounds specifically affects the CNS, while the kidney is the target organ for inorganic Hg compounds.

In this study, mercury chloride (HgCl2) was studied in a renal derived cell system, i.e., the tubular epithelial Madin-Darby canine kidney (MDCK) cell line, which has specific sensitivity to the toxic effect of mercury. MDCK cells were cultured for 6-24 hr in vitro in various concentrations (0.1-100 M) of HgCl2, and the markers of apoptosis or cell death were assayed, including DNA fragmentation, caspase-3 activity andwestern blotting of cytochrome c. The influence of the metal on cell proliferation and viability were evaluated by the conventional MTT test.

The cell viability was decreased in a time and concentration dependent fashion: decreases were noted at 6, 12 and 24 hr after HgCl2 exposure. The increases of DNA fragmentation were also observed in the concentrations from 0.1 to 10 M of HgCl2 at 6 hr after exposure. However, we could not observe DNA fragmentation in the concentrations more than 25 M because the cells rapidly proceeded to necrotic cell death. The activation of caspase-3 was also observed at 6 hr exposure in the HgCl2 concentrations from 0.1 to 10 M. The release of cytochrome c from the mitochondria into the cytosol, which is an initiator of the activation of the caspase cascade, was also observed in the HgCl2-treated MDCK cells.

These results suggest that the activation of caspase-3 was involved in HgCl2-induced apoptosis. The release of cytochrome c from the mitochondria into the cytosol was also observed in the HgCl2-treated MDCK cells. These findings indicate that in MDCK cells, HgCl2 is a potent inducer of apoptosis via cytochrome c release from the mitochondria.