Unit

UNIT 4.35 Manipulation of Expression of Arsenic (+3 Oxidation State) Methyltransferase in Cultured Cells

  1. Zuzana Drobna1,
  2. Miroslav Styblo1,2,
  3. David J. Thomas3

Published Online: 1 FEB 2010

DOI: 10.1002/0471140856.tx0435s43

Current Protocols in Toxicology

Current Protocols in Toxicology

How to Cite

Drobna, Z., Styblo, M. and Thomas, D. J. 2010. Manipulation of Expression of Arsenic (+3 Oxidation State) Methyltransferase in Cultured Cells. Current Protocols in Toxicology. 43:4.35:4.35.1–4.35.24.

Author Information

  1. 1

    University of North Carolina at Chapel Hill, Chapel Hill, North Carolina

  2. 2

    Center for Environmental Medicine, Asthma, and Lung Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina

  3. 3

    Pharmacokinetics Branch, Experimental Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina

Publication History

  1. Published Online: 1 FEB 2010
  2. Published Print: FEB 2010

Abstract

Methylation of inorganic arsenic to produce mono-, di-, or trimethylated products is the central process in the cellular metabolism of arsenic. Identification of arsenic (+3 oxidation state) methyltransferase (As3mt) as the enzyme that could catalyze all the steps in the pathway for arsenic methylation suggests that expression of this enzyme could be a useful target for manipulation. Here, methods are described for heterologous expression of the rat As3mt gene in a human urothelial cell line that normally does not express this enzyme and for silencing of the AS3MT gene by RNA interference in a human hepatoma cell line. These tools can be applied to elucidating the role of methylation in the toxic and carcinogenic effects of arsenicals. Curr. Protoc. Toxicol. 43:4.35.1-4.35.24. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • arsenic;
  • arsenic (+3 oxidation state) methyltransferase;
  • UROtsa cells (human urothelial cells);
  • heterologous expression;
  • human hepatoma cells (HepG2 cells);
  • RNA interference;
  • shRNA