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[Establishment of transgenic cell line CHL-3A4 and its metabolic activation].

作者信息

Chen Q, Wu J, Yu Y

机构信息

Institute for Hematology, First Hospital Affiliated to Zhejiang Medical University, Hangzhou.

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 1998 Sep;32(5):281-4.

PMID:10322772
Abstract

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.

摘要

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Establishment of a transgenic cell line stably expressing human cytochrome P450 2C18 and identification of a CYP2C18 clone with exon 5 missing.
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