Expression of drug metabolizing enzymes in hepatocyte-like cells derived from human embryonic stem cells
Expression of drug metabolizing enzymes in hepatocyte-like cells derived from human embryonic stem cells
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DOI：
10.1016/j.bcp.2007.05.009
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发表时间：
2007-08-01
影响因子：
5.8
通讯作者：
Johansson, Inger
中科院分区：
文献类型：
--
作者：
Ek, Monica;Soderdahl, Therese;Johansson, Inger
人胚胎干细胞（hESC）为功能性人肝细胞提供了潜在的无限来源，因为它们可以分化为显示特征性肝形态并表达几种肝标志物的肝细胞样细胞。这样的细胞可用于例如药物代谢和肝毒性的研究，然而这将需要药物代谢酶的显著表达。因此，我们研究了细胞色素P450（CYP），UDP-葡萄糖醛酸转移酶（UGT），药物转运蛋白，转录因子和其他肝脏特异性基因的表达在肝细胞样细胞来源于hESC使用一个简单的直接分化协议。采用低密度芯片、真实的时间PCR和蛋白质印迹法检测几种重要CYP的mRNA和蛋白质表达。在来自两种不同的hESC系之一的肝细胞样细胞中检测到mRNA水平上的显着表达，其远高于未分化的hESC，并且通常高于HepG2细胞。在两个品系中均检测到CYPIA2、CYP3A4/7和低水平的CYP1A1和CYP2C8/9/19蛋白。各种CYP和肝脏特异性因子的mRNA在两种细胞系中均显示为可诱导的，这反映在CYP 1A2和CYP 3A4/7蛋白的诱导水平上。这是第一份关于hESC衍生的肝细胞样细胞中所有主要CYP表达的报告，代表了迈向功能性肝细胞的重要一步，但在它们表现出与原代人肝细胞和肝脏相似的药物代谢酶水平之前，需要努力使用优化的方案进一步分化细胞。(c)2007年由Elsevier Inc.出版
Human embryonic stem cells (hESC) offer a potential unlimited source for functional human hepatocytes, since they can differentiate into hepatocyte-like cells displaying a characteristic hepatic morphology and expressing several hepatic markers. Such cells could be used for, e.g. studies of drug metabolism and hepatotoxicity, which however would require a significant expression of drug metabolising enzymes. Thus, we have investigated the expression of cytochrome P450s (CYPs), UDP-glucuronosyltransferases (UGTs), drug transporters, transcription factors and other liver specific genes in hepatocyte-like cells derived from hESC using a simple direct differentiation protocol. The mRNA and protein expression of several important CYPs were determined using low density arrays, real time PCR and Western blotting. Significant CYP expression on the mRNA level was detected in hepatocyte-like cells derived from one out of two different hESC lines tested, which was much higher than in undifferentiated hESC and generally higher than in HepG2 cells. CYPIA2, CYP3A4/7 and low levels of CYP1A1 and CYP2C8/9/19 protein were detected in both lines. The mRNAs for a variety of CYPs and liver specific factors were shown to be inducible in both cell lines, and this was reflected in induced levels of CYP1A2 and CYP3A4/7 protein. This first report on expression of all major CYPs in hepatocyte-like cells derived from hESC represents an important step towards functional hepatocytes, but efforts to further differentiate the cells using optimized protocols are needed before they exhibit similar levels of drug metabolizing enzymes as primary human hepatocytes and liver. (c) 2007 Published by Elsevier Inc.