UNIT 4.35 Manipulation of Expression of Arsenic (+3 Oxidation State) Methyltransferase in Cultured Cells
Published Online: 1 FEB 2010
Copyright © 2010 by John Wiley & Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 FEB 2010
- Published Print: FEB 2010
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.
- arsenic (+3 oxidation state) methyltransferase;
- UROtsa cells (human urothelial cells);
- heterologous expression;
- human hepatoma cells (HepG2 cells);
- RNA interference;