Jouan Elodie, Le Vée Marc, Denizot Claire, Parmentier Yannick, Fardel Olivier
Institut de Recherches en Santé, Environnement et Travail (IRSET), UMR INSERM U1085, Faculté de Pharmacie, 2 Avenue du Pr Léon Bernard, 35043 Rennes, France.
Centre de Pharmacocinétique, Technologie Servier, 25-27 Rue Eugène Vignat, 45000 Orléans, France.
Pharmaceutics. 2016 Dec 28;9(1):3. doi: 10.3390/pharmaceutics9010003.
Human hepatoma cells may represent a valuable alternative to the use of human hepatocytes for studying hepatic drug transporters, which is now a regulatory issue during drug development. In the present work, we have characterized hepatic drug transporter expression, activity and regulation in human hepatoma HuH-7 cells, in order to determine the potential relevance of these cells for drug transport assays. HuH-7 cells displayed notable multidrug resistance-associated protein (MRP) activity, presumed to reflect expression of various hepatic MRPs, including MRP2. By contrast, they failed to display functional activities of the uptake transporters sodium taurocholate co-transporting polypeptide (NTCP), organic anion-transporting polypeptides (OATPs) and organic cation transporter 1 (OCT1), and of the canalicular transporters P-glycoprotein and breast cancer resistance protein (BCRP). Concomitantly, mRNA expressions of various sinusoidal and canalicular hepatic drug transporters were not detected (NTCP, OATP1B1, organic anion transporter 2 (OAT2), OCT1 and bile salt export pump) or were found to be lower (OATP1B3, OATP2B1, multidrug and toxin extrusion protein 1, BCRP and MRP3) in hepatoma HuH-7 cells than those found in human hepatocytes, whereas other transporters such as OAT7, MRP4 and MRP5 were up-regulated. HuH-7 cells additionally exhibited farnesoid X receptor (FXR)- and nuclear factor erythroid 2-related factor 2 (Nrf2)-related up-regulation of some transporters. Such data indicate that HuH-7 cells, although expressing rather poorly some main hepatic drug transporters, may be useful for investigating interactions of drugs with MRPs, notably MRP2, and for studying FXR- or Nrf2-mediated gene regulation.
在药物研发过程中，由于人类肝细胞的使用目前已成为一个监管问题，因此人类肝癌细胞可能是用于研究肝脏药物转运体的一种有价值的替代选择。在本研究中，我们对人类肝癌HuH-7细胞中肝脏药物转运体的表达、活性及调控进行了表征，以确定这些细胞在药物转运测定中的潜在相关性。HuH-7细胞表现出显著的多药耐药相关蛋白（MRP）活性，推测这反映了包括MRP2在内的各种肝脏MRP的表达。相比之下，它们未能表现出摄取转运体牛磺胆酸钠共转运多肽（NTCP）、有机阴离子转运多肽（OATP）和有机阳离子转运体1（OCT1）以及胆小管转运体P-糖蛋白和乳腺癌耐药蛋白（BCRP）的功能活性。同时，未检测到各种肝血窦和胆小管肝脏药物转运体的mRNA表达（NTCP、OATP1B1、有机阴离子转运体2（OAT2）、OCT1和胆盐输出泵），或者发现肝癌HuH-7细胞中这些转运体的mRNA表达低于人类肝细胞中的表达（OATP1B3、OATP2B1、多药和毒素外排蛋白1、BCRP和MRP3），而其他转运体如OAT7、MRP4和MRP5则上调。HuH-7细胞还表现出法尼醇X受体（FXR）和核因子红细胞2相关因子2（Nrf2）相关的一些转运体上调。这些数据表明，HuH-7细胞虽然某些主要肝脏药物转运体的表达相当低，但可能有助于研究药物与MRP（尤其是MRP2）的相互作用，以及研究FXR或Nrf2介导的基因调控。
