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Updated: Aug 9, 2026
Mass Spectrometry and Luminogenic-based Approaches to Characterize Phase I Metabolic Competency of In Vitro Cell Cultures
Published on: March 28, 2017
[Establishment of transgenic cell line CHL-3A4 and its metabolic activation]
1Institute 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.
