Mayati Abdullah, Moreau Amélie, Jouan Elodie, Febvre-James Marie, Denizot Claire, Parmentier Yannick, Fardel Olivier
Univ Rennes, Inserm, EHESP, IRSET (Institut de Recherche en Santé, Environnement et Travail) - UMR_S 1085, F-35000 Rennes, France.
Centre de Pharmacocinétique, Technologie Servier, F-45000 Orléans, France.
Pharmaceutics. 2018 Nov 21;10(4):246. doi: 10.3390/pharmaceutics10040246.
The HepaRG cell line is a highly differentiated human hepatoma cell line, displaying the expression of various drug transporters. However, functional expression of nucleoside transporters remains poorly characterized in HepaRG cells, although these transporters play a key role in hepatic uptake of antiviral and anticancer drugs. The present study was, therefore, designed to characterize the expression, activity and regulation of equilibrative (ENT) and concentrative (CNT) nucleoside transporter isoforms in differentiated HepaRG cells. These cells were found to exhibit a profile of nucleoside transporter mRNAs similar to that found in human hepatocytes, i.e., notable expression of ENT1, ENT2 and CNT1, with very low or no expression of CNT2 and CNT3. ENT1 activity was, next, demonstrated to be the main uridine transport activity present in HepaRG cells, like in cultured human hepatocytes. Various physiological factors, such as protein kinase C (PKC) activation or treatment by inflammatory cytokines or hepatocyte growth factor (HGF), were additionally found to regulate expression of ENT1, ENT2 and CNT1; PKC activation and HGF notably concomitantly induced mRNA expression and activity of ENT1 in HepaRG cells. Overall, these data suggest that HepaRG cells may be useful for analyzing cellular pharmacokinetics of nucleoside-like drugs in human hepatic cells, especially of those handled by ENT1.
HepaRG细胞系是一种高度分化的人肝癌细胞系，可表达多种药物转运蛋白。然而，核苷转运蛋白的功能表达在HepaRG细胞中仍未得到充分表征，尽管这些转运蛋白在肝脏摄取抗病毒和抗癌药物中起关键作用。因此，本研究旨在表征分化的HepaRG细胞中平衡型（ENT）和浓缩型（CNT）核苷转运蛋白亚型的表达、活性和调节。发现这些细胞呈现出与人类肝细胞中相似的核苷转运蛋白mRNA谱，即ENT1、ENT2和CNT1显著表达，而CNT-2和CNT-3表达极低或不表达。接下来，ENT1活性被证明是HepaRG细胞中主要的尿苷转运活性，就像在培养的人类肝细胞中一样。此外，还发现各种生理因素，如蛋白激酶C（PKC）激活或炎性细胞因子或肝细胞生长因子（HGF）处理，可调节ENT1、ENT2和CNT1的表达；PKC激活和HGF显著地同时诱导HepaRG细胞中ENT1的mRNA表达和活性。总体而言，这些数据表明HepaRG细胞可能有助于分析核苷类药物在人肝细胞中的细胞药代动力学，特别是由ENT1处理的药物。
