[Establishment of transgenic cell line CHL-3A4 and its metabolic activation]
[Article in Chinese]
Institute for Hematology, First Hospital Affiliated to Zhejiang Medical University, Hangzhou.
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). CONCLUSION: 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.
PMID: 10322772 [PubMed - indexed for MEDLINE]