| Publication number |
CLPUB00804 |
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| Authors |
Zheng-Kai Xue, Hong Wei, Hai-Tao Shang, Gen-Qing Yang, Bin Ye; |
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| Title |
Establishment of a HepG2 cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs. |
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| Citation |
Di 3 Jun Yi Da Xue Xue Bao 31:1552-1555(2009) |
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| Web pages |
https://oversea.cnki.net/KCMS/detail/detail.aspx?dbcode=CJFD&dbname=CJFD2009&filename=DSDX200916015&uniplatform=OVERSEA |
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| Abstract |
Objective: To establish a HepG2 cell line, which can stably express Sus
scrofa CYP3A22,for the evaluation of the drug metabolic characteristics of
CYP3A22.
Methods: Sus scrofa CYP3A22 gene obtained by PCR was cloned into the
eukaryotic expression vector pcDNA3.1 [The tissue of Bama minipig was
amplified by RT-PCR. The gene was subcloned into plasmid pMD18-T
(designated as pMD-3A22), and then the gene amplified by recombinant
plasmid was designated as pcDNA-3A22]. Then Sus scrofa CYP3A22 gene was
confirmed by sequencing. Next,the confirmed gene was transfected into HepG2
cells by lipid-media transfection,and the transformant HepG2 cells were
screened by G418 for 10 generations. Expression of CYP3A22 in HepG2 cells
was identified by RT-PCR and Western blot analyses,and the metabolic
activity of transformant HepG2 (HepG2-CYP3A22) was verified by
nifedipine oxidation.
Results: The HepG2-CYP3A22 line demonstrated a significantly higher
oxidative activity as compared with the negative control HepG2 line.
Conclusion HepG2 cell line stably expressing CYP3A22 was established
successfully. The cell line may be useful for the studies of toxicity and
pharmacology.
|
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| Cell lines |
CVCL_E8Z3; HepG2-CYP3A22 |
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