[Establishment of transgenic cell line CHL-3A4 and its metabolic activation].
OBJECTIVE
To establish a model cell line CHL-3A4 to study metabolic pathways for some genotoxic chemicals.


METHODS
Complimentary DNA (cDNA) of cloned cytochrome P450-3A4 gene in human liver tissue was transferred to Bluescript M13 vector with reverse transcription polymerase chain reaction (RT-PCR) and DNA recombinant techniques. Restriction endonuclease map analysis and sequencing of partial cloned fragment proved that CYP3A4 cDNA was cloned into Bluescript M13 vector. Then, recombinant expression plasmid pREP9-3A4 was constructed in eukaryotic cell and transfected into Chinese hamster CHL cells.


RESULTS
It was proved that a CHL-3A4 transgenic cell line was established, which could lead metabolic activation for aflatoxin B1 (AFB1), sterigmatocystin (STC) and cyclophosphamide (CPA).


CONCLUSIONS
The CHL-3A4 cell line established did express human cytochrome P450 3A4 and could lead metabolic activation for three genotoxic chemicals AFB1, STC and CPA.
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D018076:DNA, Complementary; D005347:Fibroblasts / Q000166:cytology; D018014:Gene Transfer Techniques; D008099:Liver / Q000378:metabolism*; D008168:Lung / Q000166:cytology; D010957:Plasmids; D019076:Transgenes