Abstract

1. The induction of cytochrome P450s can result in reduced drug efficacy and lead to potential drug–drug interactions. The xenoreceptors—aryl hydrocarbon receptor (AhR), constitutive androstane receptor (CAR), and pregnane X receptor (PXR)—play key roles in CYP induction by xenobiotics. In order to be able to rapidly screen for the induction of three enzymes (CYP1A1, CYP2B6, and CYP3A4), we generated a stable AhR-responsive HepG2 cell line, a stable CAR-responsive HepG2 cell line, and a stable PXR-responsive HepG2 cell line.

2. To validate these stable xenoreceptor-responsive HepG2 cell lines, we evaluated the induction of the different Gaussia reporter activities, as well as the mRNA and protein expression levels of endogenous CYPs in response to different inducers.

3. The induction of luciferase activity in the stable xenoreceptor-responsive HepG2 cell lines by specific inducers occurred in a concentration dependent manner. There was a positive correlation between the induction of luciferase activities and the induction endogenous CYP mRNA expression levels. These xenoreceptor-responsive HepG2 cell lines were further validated with known CYP1A1, CYP2B6, and CYP3A4 inducers.

4. These stable xenoreceptor-responsive HepG2 cell lines may be used in preclinical research for the rapid and sensitive detection of AhR, CAR, and PXR ligands that induce CYP450 isoforms.

摘要

1. 细胞色素 P450 的诱导会导致药物疗效降低并导致潜在的药物相互作用。异种受体——芳烃受体 (AhR)、组成型雄甾烷受体 (CAR) 和孕烷 X 受体 (PXR)——在外源性物质的 CYP 诱导中起关键作用。为了能够快速筛选三种酶（CYP1A1、CYP2B6 和 CYP3A4）的诱导，我们生成了稳定的 AhR 反应性 HepG2 细胞系、稳定的 CAR 反应性 HepG2 细胞系和稳定的 PXR 反应性 HepG2细胞系。

2. 为了验证这些稳定的异种受体反应性 HepG2 细胞系，我们评估了不同 Gaussia 报告基因活性的诱导，以及响应不同诱导剂的内源性 CYP 的 mRNA 和蛋白质表达水平。

3. 在稳定的异种受体反应性 HepG2 细胞系中，特定诱导剂对荧光素酶活性的诱导以浓度依赖性方式发生。荧光素酶活性的诱导与诱导内源性CYP mRNA表达水平之间存在正相关。这些异种受体反应性 HepG2 细胞系用已知的 CYP1A1、CYP2B6 和 CYP3A4 诱导剂进一步验证。

4. 这些稳定的异种受体反应性 HepG2 细胞系可用于临床前研究，以快速灵敏地检测诱导 CYP450 亚型的 AhR、CAR 和 PXR 配体。