Department of Pharmaceutical Sciences, School of Pharmacy (L.C., Y.B., S.C.P., X.-b.Z.), and Department of Physiology and Neurobiology (K.Z.), University of Connecticut, Storrs, Connecticut; and Department of Pharmacology, School of Basic Medicine, Zhengzhou University, Zhengzhou, Henan, China (L.Z.).
Department of Pharmaceutical Sciences, School of Pharmacy (L.C., Y.B., S.C.P., X.-b.Z.), and Department of Physiology and Neurobiology (K.Z.), University of Connecticut, Storrs, Connecticut; and Department of Pharmacology, School of Basic Medicine, Zhengzhou University, Zhengzhou, Henan, China (L.Z.)
Mol Pharmacol. 2018 Jul;94(1):749-759. doi: 10.1124/mol.118.112235. Epub 2018 Apr 24.
Cytochrome P450 (P450) enzymes are responsible for metabolizing drugs. Expression of P450s can directly affect drug metabolism, resulting in various outcomes in therapeutic efficacy and adverse effects. Several nuclear receptors are transcription factors that can regulate expression of P450s at both basal and drug-induced levels. Some long noncoding RNAs (lncRNAs) near a transcription factor are found to participate in the regulatory functions of the transcription factors. The aim of this study is to determine whether there is a transcriptional regulatory network containing nuclear receptors and lncRNAs controlling both basal and drug-induced expression of P450s in HepaRG cells. Small interfering RNAs or small hairpin RNAs were applied to knock down four nuclear receptors [hepatocyte nuclear factor 1 (HNF1), hepatocyte nuclear factor 4 (HNF4), pregnane X receptor (PXR), and constitutive androstane receptor (CAR)] as well as two lncRNAs [HNF1 antisense RNA 1 (HNF1-AS1) and HNF4 antisense RNA 1 (HNF4-AS1)] in HepaRG cells with or without treatment of phenobarbital or rifampicin. Expression of eight P450 enzymes was examined in both basal and drug-induced levels. CAR and PXR mainly regulated expression of specific P450s. HNF1 and HNF4 affected expression of a wide range of P450s as well as other transcription factors. HNF1 and HNF4 controlled the expression of their neighborhood lncRNAs, HNF1-AS1 and HNF4-AS1, respectively. HNF1-AS1 and HNF4-AS1 was also involved in the regulation of P450s and transcription factors in diverse manners. Altogether, our study concludes that a transcription regulatory network containing the nuclear receptors and lncRNAs controls both basal and drug-induced expression of P450s in HepaRG cells.
细胞色素 P450（P450）酶负责代谢药物。P450 的表达可以直接影响药物代谢，从而导致治疗效果和不良反应的各种结果。几种核受体是转录因子，可以在基础水平和药物诱导水平上调节 P450 的表达。在转录因子附近发现一些长非编码 RNA（lncRNA）参与转录因子的调节功能。本研究旨在确定是否存在一个包含核受体和 lncRNA 的转录调控网络，控制 HepaRG 细胞中 P450 的基础和药物诱导表达。小干扰 RNA 或短发夹 RNA 被应用于敲低四个核受体[肝细胞核因子 1（HNF1）、肝细胞核因子 4（HNF4）、孕烷 X 受体（PXR）和组成型雄烷受体（CAR）]以及两个 lncRNA[HNF1 反义 RNA 1（HNF1-AS1）和 HNF4 反义 RNA 1（HNF4-AS1）]在有或没有苯巴比妥或利福平处理的 HepaRG 细胞中。在基础和药物诱导水平上检查了 8 种 P450 酶的表达。CAR 和 PXR 主要调节特定 P450 的表达。HNF1 和 HNF4 影响广泛的 P450 以及其他转录因子的表达。HNF1 和 HNF4 分别控制其邻近 lncRNA，HNF1-AS1 和 HNF4-AS1 的表达。HNF1-AS1 和 HNF4-AS1 也以不同的方式参与 P450 和转录因子的调节。总之，我们的研究得出结论，一个包含核受体和 lncRNA 的转录调控网络控制 HepaRG 细胞中 P450 的基础和药物诱导表达。
