Hepatocytes from metallothionein-I and II knock-out mice are sensitive to cadmium- and tert-butylhydroperoxide-induced cytotoxicity.

Metallothionein (MT) has been proposed to play an important role in heavy metal detoxication and in the scavenging of free radicals. Effects of MT on the cytotoxicity of cadmium (Cd), tert-butylhydroperoxide (t-BHP) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were examined using primary hepatocyte cultures from control (C57BL/6J) and MT-I and II knock-out (MT-null) mice. Compared to control hepatocytes, MT-null hepatocytes had minimal Cd-binding proteins (MT equivalents), but cellular glutathione concentration was similar to the control hepatocytes. MT-null hepatocytes were more sensitive than controls to the cytotoxic effects of Cd (50-300 microM) and t-BHP (125-500 microM), as indicated by the levels of lactate dehydrogenase released into the medium. Cd and t-BHP also produced more lipid peroxidation in MT-null hepatocytes than in control cells, as demonstrated by the abundance of thiobarbituric acid-reactive substances. However, MT-null hepatocytes were equally sensitive as controls to the cytotoxicity of MNNG (0.5-2.0 mM), suggesting that MT does not protect against MNNG-induced cytotoxicity. These results support the hypothesis that constitutive MT levels affect the sensitivity of mammalian cells to Cd and oxidative stress.