This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Nouso, K. Articles by Battula, N. Search for Related Content PubMed PubMed Citation Articles by Nouso, K. Articles by Battula, N.

Stable Expression of Human Cytochrome P450IIE1 in Mammalian Cells: Metabolic Activation of Nitrosodimethylamine and Formation of Adducts with Cellular DNA¹

Laboratory of Experimental Carcinogenesis, National Cancer Institute, Bethesda, Maryland 20892

To introduce cytochrome P450IIE1 DNA stably into the chromosomal DNA of mammalian cells, we constructed recombinant retroviruses containing the full-length complementary DNA for human cytochrome P450IIE1 and a selectable neo gene. Rat and mouse cells were infected with these viruses, and clones expressing the neo marker gene product were selected in G418-containing medium. Analysis of the DNA of the clones by Southern blotting showed that the viral DNA was integrated into the cellular DNA. Enzymatic analysis of the clones showed that the transduced DNA directed the expression of enzymatically active cytochrome P450IIE1. Treatment of the cells with the carcinogen [¹⁴C]-nitrosodimethylamine and analysis of the cellular DNA by CsCl equilibrium density gradients showed incorporation of the label into DNA, indicating the formation of covalent adducts with the cell DNA. Construction of recombinant cell lines constitutively expressing cytochrome P450IIE1 provides a permanent source for this enzyme and can aid in the analysis of its catalytic properties, such as the metabolic activation of chemical mutagens/carcinogens, and the consequent cytotoxic and genotoxic effects of these compounds.

¹ K. N. was supported by a Japanese Overseas Cancer Fellowship of the Foundation for Promotion of Cancer Research.

² To whom reprint requests should be addressed, at Laboratory of Experimental Carcinogenesis, National Cancer Institute, Building 37, Room 3C28, Bethesda, MD 20892.

³ Present address: Division of Antiviral Drug Products, FDA, HFD-530, Kensington, MD 20895.

Received 10/14/91. Accepted 1/23/92.

This article has been cited by other articles:

				C. Chen, L. S. Cook, X.-Y. Li, S. Hallagan, M. M. Madeleine, J. R. Daling, and N. S. Weiss CYP2D6 Genotype and the Incidence of Anal and Vulvar Cancer1 Cancer Epidemiol. Biomarkers Prev., April 1, 1999; 8(4): 317 - 321. [Abstract] [Full Text] [PDF]

				J.-Y. Huan and D. R. Koop Tightly Regulated and Inducible Expression of Rabbit Cyp2e1 Using A Tetracycline-Controlled Expression System Drug Metab. Dispos., April 1, 1999; 27(4): 549 - 554. [Abstract] [Full Text]