Archives of Toxicology

, Volume 87, Issue 2, pp 311–321 | Cite as

Metallothionein blocks oxidative DNA damage in vitro



The role of metallothionein (MT) in mitigation of oxidative DNA damage (ODD) induced by either cadmium (Cd) or the direct oxidant hydrogen peroxide (H2O2) was systematically examined using MT-I/II double knockout (MT-null) or MT-competent wild-type (WT) cells. Both toxicants were much more lethal to MT-null cells (Cd LC50 = 6.6 μM; H2O2 LC50 = 550 μM) than to WT cells (Cd LC50 = 16.5 μM; H2O2 LC50 = 930 μM). Cd induced concentration-related MT increases in WT cells, while the basal levels were undetectable and not increased by Cd in MT-null cells. ODD, measured by the immuno-spin trapping method, was minimally induced by sub-toxic Cd levels (1 or 5 μM; 24 h) in WT cells, but markedly increased in MT-null cells (>430 %). Similarly, ODD was induced to higher levels by lower concentrations of H2O2 in MT-null cells than WT cells. Transfection of MT-I into MT-null cells reduced both Cd- and H2O2-induced cytolethality and ODD. Cd increased the expression of the oxidant defense genes, HO-1, and GSTa2 to a much greater extent in MT-null cells than in WT. Cd or H2O2 exposure increased the expression of key transport genes, Mrp1 and Mrp2, in WT cells but not in MT-null cells. MT protects against Cd- and H2O2-induced ODD in MT-competent cells possibly by multiple mechanisms, potentially including direct metal ion sequestration and sequestration of oxidant radicals by MT. MT-deficient cells appear to adapt to Cd primarily by turning on oxidant response systems, while MT-competent cells activate MT and transport systems.


DNA Cadmium Oxidative damage Metallothionein Cancer 





Cadmium chloride


5,5-dimethyl-1-pyrroline N-oxide


Glutathione S-transferase


Glutathione S-transferase pi 1


Glutathione S-transferase-α2


Heme oxygenase 1


Hydrogen peroxide


Immuno-spin trapping


Lethal concentration 50 %


Multidrug resistance-related protein 1


Multidrug resistance-related protein 2




MT-I/II knockout


Oxidative DNA damage


Reverse transcription-polymerase chain reaction


Reactive oxygen species


Wild type



The authors thank Drs. Rachel Person, Katie Pelch, John Bucher, and Nigel Walker for critical review of this manuscript. The authors also thank Matthew W. Bell for aid with the graphics. This article may be the work product of an employee or a group of employees of the NIEHS, NIH; however, the statements contained herein do not necessarily represent the statements, opinions, or conclusions of the NIEHS, NIH. or the United States Government. The content of this publication does not necessarily reflect the views or the policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.

Conflict of interest



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Copyright information

© Springer-Verlag (outside the USA) 2012

Authors and Affiliations

  1. 1.Inorganic Toxicology Group, National Toxicology Program Laboratory, Division of the National Toxicology ProgramNational Institute of Environmental Health SciencesResearch Triangle ParkUSA
  2. 2.Division of Translational BiologyThe Hamner Institutes for Health SciencesResearch Triangle ParkUSA

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