Establishment of a HepG<sub>2</sub> cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs

薛正楷; 魏泓; 商海涛; 杨根庆; 叶彬

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Establishment of a HepG₂ cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs

更新时间：2026-01-09

- Establishment of a HepG₂ cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs
- Journal of Army Medical University Vol. 31, Issue 16, Pages: 1552-1555(2009)
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  CLC： R322.47;R394-33;R394.6
- Published：2009
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薛正楷,魏泓,商海涛,杨根庆,叶彬.CYP3A22稳定表达HepG2细胞株的建立及其探针药物代谢活性鉴定[J].陆军军医大学学报(原第三军医大学学报),2009,31(16):1552-1555.

　XUE Zheng-kai,WEI Hong,SHANG Hai-tao,YANG Gen-qing,YE Bin.Establishment of a HepG2 cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(16):1552-1555.
薛正楷,魏泓,商海涛,杨根庆,叶彬.CYP3A22稳定表达HepG2细胞株的建立及其探针药物代谢活性鉴定[J].陆军军医大学学报(原第三军医大学学报),2009,31(16):1552-1555. DOI：

　XUE Zheng-kai,WEI Hong,SHANG Hai-tao,YANG Gen-qing,YE Bin.Establishment of a HepG2 cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(16):1552-1555. DOI：

摘要

目的建立稳定表达Bama小型猪细胞色素CYP3A22的HepG

细胞株。方法通过从Bama小型猪肝组织中提取tRNA、经RT-PCR得到Bama小型猪CYP3A22的基因，并将此基因克隆至亚克隆载体pMD18-T上得到重组质粒pMD-3A22；以测序确定的重组质粒pMD-3A22为模板，采用PCR扩增CYP3A22基因并在其3′添加组氨酸标签，扩增产物经BamHⅠ/ XhoⅠ双酶切，将带有组氨酸标签的CYP3A22基因定向克隆至pcDNA3.1（+）中，测序表明，CYP3A22基因真核表达载体正确构建;将测序正确的CYP3A22基因通过脂质体转染至HepG

细胞，G418筛选10代，RT-PCR及Western blot分析，并用探针药物硝苯地平对重组细胞进行进行活性鉴定。结果与HepG

相比，HepG

-CYP3A22细胞株具有极显著的硝苯地平氧化活性。结论成功建立了稳定表达CYP3A22的HepG

细胞株，可用于相关的药物代谢研究。

Abstract

Objective To establish a HepG

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 HepG

cells by lipid-media transfection

and the transformant HepG

cells were screened by G418 for 10 generations. Expression of CYP3A22 in HepG

cells was identified by RT-PCR and Western blot analyses

and the metabolic activity of transformant HepG

(HepG

-CYP3A22) was verified by nifedipine oxidation. Results The HepG

-CYP3A22 line demonstrated a significantly higher oxidative activity as compared with the negative control HepG

line. Conclusion HepG

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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Establishment of a HepG2 cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs

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摘要

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Establishment of a HepG₂ cell line stably expressing CYP3A22 and identification of the metabolic activity of the probe drugs