| Development of an in situ toxicity assay system using recombinant baculoviruses. | |
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MedLine Citation:
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PMID: 8619897 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A new method for experimentally analyzing the role of enzymes involved in metabolizing mutagenic, carcinogenic, or cytotoxic chemicals is described. Spodoptera fugiperda (SF-21) cells infected with recombinant baculoviruses are used for high level expression of one or more cloned enzymes. The ability of these enzymes to prevent or enhance the toxicity of drugs and xenobiotics is then measured in situ. Initial parameters for the system were developed and optimized using baculoviruses engineered for expression of the mouse soluble epoxide hydrolase (msEH, EC 3.3.2.3) or the rat cytochrome P4501A1. SF-21 cells expressing msEH were resistant to trans-stilbene oxide toxicity as well as several other toxic epoxides including: cis-stilbene oxide, 1,2,7,8-diepoxyoctane, allylbenzene oxide, and estragole oxide. The msEH markedly reduced DNA and protein adduct formation in SF-21 cells exposed to [3H]allylbenzene oxide or [3H]estragole oxide. On the other hand, 9,10-epoxyoctadecanoic acid and methyl 9,10-epoxyoctadecanoate were toxic only to cells expressing sEH, suggesting that the corresponding fatty acid diols were cytotoxic. This was confirmed by showing that chemically synthesized diols of these fatty acid epoxides were toxic to control SF-21 cells at the same concentration as were the epoxides to cells expressing sEH. A recombinant baculovirus containing a chimeric cDNA formed between the rat P4501A1 and the yeast NADPH-P450 reductase was also constructed and expressed in this system. A model compound, naphthalene, was toxic to SF-21 infected with the rat P4501A1/reductase chimeric co-infecting SF-21 cells with either a human or a rat microsomal EH virus along with P4501A1/reductase virus. These results demonstrate the usefulness of this new system for experimentally analyzing the role of enzymes hypothesized to metabolize endogenous and exogenous chemicals of human health concern. |
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Authors:
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D F Grant; J F Greene; F Pinot; B Borhan; M F Moghaddam; B D Hammock; B McCutchen; H Ohkawa; G Luo; T M Guenthner |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Biochemical pharmacology Volume: 51 ISSN: 0006-2952 ISO Abbreviation: Biochem. Pharmacol. Publication Date: 1996 Feb |
Date Detail:
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Created Date: 1996-06-13 Completed Date: 1996-06-13 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 0101032 Medline TA: Biochem Pharmacol Country: ENGLAND |
Other Details:
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Languages: eng Pagination: 503-15 Citation Subset: IM |
Affiliation:
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Department of Entomology and Environmental Toxicology, University of California, Davis 95616, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Baculoviridae Base Sequence Carcinogens / toxicity* Cell Line Cell Survival / drug effects Cloning, Molecular Cytochrome P-450 Enzyme System / biosynthesis, metabolism* Cytotoxins / toxicity* DNA Primers Drug Evaluation, Preclinical / methods Drug Resistance Epoxide Hydrolases / biosynthesis, metabolism* Humans Mice Molecular Sequence Data Mutagens / toxicity* Polymerase Chain Reaction Rats Recombinant Fusion Proteins / biosynthesis, metabolism Recombinant Proteins / biosynthesis, metabolism Restriction Mapping Spodoptera Structure-Activity Relationship Toxicity Tests / methods* Xenobiotics / toxicity |
| Grant Support | |
ID/Acronym/Agency:
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P30 ES05707/ES/NIEHS NIH HHS; P42 ES04699/ES/NIEHS NIH HHS; R01 ES02710/ES/NIEHS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Carcinogens; 0/Cytotoxins; 0/DNA Primers; 0/Mutagens; 0/Recombinant Fusion Proteins; 0/Recombinant Proteins; 0/Xenobiotics; 9035-51-2/Cytochrome P-450 Enzyme System; EC 3.3.2.-/Epoxide Hydrolases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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