Braeuning Albert, Thomas Maria, Hofmann Ute, Vetter Silvia, Zeller Eva, Petzuch Barbara, Johänning Janina, Schroth Werner, Weiss Thomas S, Zanger Ulrich M, Schwarz Michael
Department of Food Safety, Federal Institute for Risk Assessment, Berlin, Germany (A.B.); Dr.-Margarethe-Fischer-Bosch-Institute for Clinical Pharmacology, Stuttgart, and University of Tübingen, Tübingen, Germany (M.T., U.H., J.J., W.S., U.M.Z.); Department of Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Tübingen, Germany (A.B., S.V., E.Z., B.P., M.S.); Regensburg University Hospital, Regensburg, Germany (T.S.W.)
Department of Food Safety, Federal Institute for Risk Assessment, Berlin, Germany (A.B.); Dr.-Margarethe-Fischer-Bosch-Institute for Clinical Pharmacology, Stuttgart, and University of Tübingen, Tübingen, Germany (M.T., U.H., J.J., W.S., U.M.Z.); Department of Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Tübingen, Germany (A.B., S.V., E.Z., B.P., M.S.); Regensburg University Hospital, Regensburg, Germany (T.S.W.).
Drug Metab Dispos. 2015 Nov;43(11):1781-7. doi: 10.1124/dmd.115.064667. Epub 2015 Aug 26.
Enzymatic conversion of most xenobiotic compounds is accomplished by hepatocytes in the liver, which are also an important target for the manifestation of the toxic effects of foreign compounds. Most cell lines derived from hepatocytes lack important toxifying or detoxifying enzymes or are defective in signaling pathways that regulate expression and activity of these enzymes. On the other hand, the use of primary human hepatocytes is complicated by scarce availability of cells and high interdonor variability. Thus, analyses of drug metabolism and hepatotoxicity in vitro are a difficult task. The cell line HC-AFW1 was isolated from a pediatric hepatocellular carcinoma and so far has been used for tumorigenicity and chemotherapy resistance studies. Here, a comprehensive characterization of xenobiotic metabolism in HC-AFW1 cells is presented along with studies on the functionality of the most important transcriptional regulators of drug-metabolizing enzymes. Results from HC-AFW1 cells were compared with commercially available HepaRG cells and cultured primary human hepatocytes. Data show that the nuclear receptors and xenosensors AHR (aryl hydrocarbon receptor), CAR (constitutive androstane receptor), PXR (pregnane-X-receptor), NRF2 [nuclear factor (erythroid-derived 2)-like 2], and PPARα (peroxisome proliferator-activated receptor α) are functional in HC-AFW1 cells, comparable to HepaRG and primary cells. HC-AFW1 cells possess considerable activities of different cytochrome P450 enzymes, which, however, are lower than corresponding enzyme activities in HepaRG cells or primary hepatocytes. In summary, HC-AFW1 are a new promising tool for studying the mechanisms of the regulation of drug metabolism in human liver cells in vitro.
大多数外源性化合物的酶促转化由肝脏中的肝细胞完成，肝细胞也是外源化合物毒性作用表现的重要靶点。大多数源自肝细胞的细胞系缺乏重要的毒性化或解毒酶，或者在调节这些酶表达和活性的信号通路中存在缺陷。另一方面，原代人肝细胞的使用因细胞供应稀缺和供体间高度变异性而变得复杂。因此，体外药物代谢和肝毒性分析是一项艰巨的任务。细胞系HC-AFW1是从一名儿童肝细胞癌中分离出来的，迄今为止已用于致瘤性和化疗耐药性研究。本文介绍了HC-AFW1细胞中外源化合物代谢的全面特征，以及对药物代谢酶最重要转录调节因子功能的研究。将HC-AFW1细胞的结果与市售的HepaRG细胞和培养的原代人肝细胞进行了比较。数据表明，核受体和外源性物质传感器芳烃受体（AHR）、组成型雄甾烷受体（CAR）、孕烷X受体（PXR）、核因子（红系衍生2）样2（NRF2）和过氧化物酶体增殖物激活受体α（PPARα）在HC-AFW1细胞中具有功能，与HepaRG细胞和原代细胞相当。HC-AFW1细胞具有不同细胞色素P450酶的相当活性，然而，这些活性低于HepaRG细胞或原代肝细胞中的相应酶活性。总之，HC-AFW1是体外研究人肝细胞中药物代谢调节机制的一种新的有前景的工具。
