| Publication number |
CLPUB00644 |
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| Authors |
Xue Z.-K., Wei H., Shang H.-T., Ye B. |
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| Title |
Establishment of a HepG2 cell line stably expressing the CYP3A29 isoenzyme and identification of its nifedipine metabolic activity. |
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| Citation |
Zhongguo Ren Shou Gong Huan Bing Xue Bao 25:640-644(2009) |
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| Web pages |
http://www.rsghb.cn/EN/Y2009/V25/I07/640 |
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| Abstract |
To establish a HepG2 cell line, which stably expressing CYP3A29 of Bama
miniature pig in order to evaluate the drug metabolic characteristics of
this isoenzyme, the gene for this system was obtained through total RNA
extraction and RT-PCR assay. The gene was subcloned into plasmid PMD18-T,
designated as pMD-CYP3A29. This gene was then amplified by PCR, and cloned
into eukaryotic expression vector pcDNA3.1(+), and the recombinant plasmid
was designated as pcDNA-CYP3A29. Sequencing was used to confirmed the
correctness of the gene of this gene. The expressed gene was then
transfected into HepG2 cells by lipid-media transfection and the
transformants were screened by G418 for 10 generations;the expression of
CYP3A29 was identified by RT-PCR- West-blot and the metabolic activity of
the transformant HepG2-CYP3A29 was verified by nifedipine oxidation. In
comparison with HepG2, the transformant HepG2-CYP3A29 showed remarkable
oxidative activity. It is apparent that the cell line stably expressing
CYP3A29 isoenzyme was successfully established, and it may be used for the
metabolic study of related drugs.
|
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| Cell lines |
CVCL_A9JD; HepG2-CYP3A29 |
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