Document Detail


Retrovirus-mediated stable expression of human CYP2A6 in mammalian cells.
MedLine Citation:
PMID:  8223969     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To study the pharmacological and toxicological significance of the human cytochrome P450 isoform CYP2A6, we expressed it in mammalian cells by retrovirus-mediated gene transfer. The LXSN vector and PA317 packaging cells were used to create amphotropic recombinant retroviruses containing CYP2A6 cDNA. NIH 3T3 and HeLa cells were infected with these retroviruses and cell clones expressing CYP2A6 were selected. The integration of the CYP2A6 construct was verified by PCR analysis and northern blot analysis showed that a 5 kb mRNA containing the CYP2A6 was present in the cells. The integrated cDNA directed the expression of catalytically active CYP2A6 enzyme which has remained stable over numerous cell passages. No oxidation of several other P450 substrates was detected. The promutagen aflatoxin B1 was metabolized to intermediates binding to the host cell genomic DNA by the 3T3 2A6 cells. These cell lines are thus well suited for the study of the catalytic profile and the biological consequences of promutagen activation by the human CYP2A6 isoform.
Authors:
P Salonpää; J Hakkola; M Pasanen; O Pelkonen; K Vähäkangas; N Battula; K Nouso; H Raunio
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  European journal of pharmacology     Volume:  248     ISSN:  0014-2999     ISO Abbreviation:  Eur. J. Pharmacol.     Publication Date:  1993 Aug 
Date Detail:
Created Date:  1993-12-21     Completed Date:  1993-12-21     Revised Date:  2014-01-09    
Medline Journal Info:
Nlm Unique ID:  1254354     Medline TA:  Eur J Pharmacol     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  95-102     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
3T3 Cells
Aflatoxin B1 / metabolism
Animals
Aryl Hydrocarbon Hydroxylases*
Base Sequence
Biotransformation
Cloning, Molecular
Cytochrome P-450 Enzyme System / antagonists & inhibitors,  biosynthesis*,  chemistry,  genetics,  metabolism
DNA, Complementary / metabolism
Gene Transfer Techniques*
Genetic Vectors
HeLa Cells
Humans
Mice
Mixed Function Oxygenases / antagonists & inhibitors,  biosynthesis*,  chemistry,  genetics,  metabolism
Molecular Sequence Data
Polymerase Chain Reaction
RNA, Messenger / analysis,  genetics
Recombinant Proteins / genetics
Retroviridae / genetics*
Chemical
Reg. No./Substance:
0/DNA, Complementary; 0/RNA, Messenger; 0/Recombinant Proteins; 9035-51-2/Cytochrome P-450 Enzyme System; 9N2N2Y55MH/Aflatoxin B1; EC 1.-/Mixed Function Oxygenases; EC 1.14.13.-/coumarin 7-hydroxylase; EC 1.14.14.1/Aryl Hydrocarbon Hydroxylases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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