Hepatic cell lines for drug hepatotoxicity testing: limitations and strategies to upgrade their metabolic competence by gene engineering.
Hepatic cell lines for drug hepatotoxicity testing: limitations and strategies to upgrade their metabolic competence by gene engineering.
复制标题
用于药物肝毒性测试的肝细胞系：通过基因工程提升其代谢能力的局限性和策略。
DOI：
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发表时间：
2013
影响因子：
2.3
通讯作者：
M. Gómez‐Lechón
中科院分区：
文献类型：
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作者：
M. Donato;R. Jover;M. Gómez‐Lechón
新化合物的药物开发中的一个关键问题是对药物的代谢、所涉及的酶和潜在肝毒性的了解。原代培养的肝细胞是研究药物代谢的重要体外模型。然而，人肝细胞显示出表型不稳定性，并且具有有限的可及性和高批次间功能变异性，这使其在常规检测中的使用严重复杂化。因此，已经开发了几种肝源性细胞模型用于药物代谢和肝毒性筛选以克服这些缺点。肝癌细胞系提供了重要的优势，可用性，无限的寿命和稳定的表型，从而使它们成为此类研究的合适模型。然而，目前可用的人肝癌细胞系不是培养肝细胞的良好替代品，因为它们显示出对大多数药物代谢酶的非常有限的表达。已经开发了其他方法来产生具有代谢能力的永生化肝细胞（使用编码永生化基因的质粒来转化人肝细胞、从转基因动物获得的细胞系、肝细胞切片机或肝细胞）。在肝癌细胞中异源表达细胞色素P450酶的重组模型也已经产生，并且广泛用于药物代谢和毒性评价。近年来，上调人细胞系中药物生物转化酶表达的新方法（即，用编码关键肝转录因子的表达载体转染）也已经进行了研究。本文综述了肝源性细胞系的特点，它们对药物代谢和肝毒性研究的适用性，以及产生代谢活性肝细胞系的最新策略。
One key issue in the pharmaceutical development of new compounds is knowledge on metabolism, the enzymes involved and the potential hepatotoxicity of a drug. Primary cultured hepatocytes are a valuable in vitro model for drug metabolism studies. However, human hepatocytes show phenotypic instability and have restricted accessibility and high batch-to-batch functional variability, which seriously complicates their use in routine testing. Therefore, several liver-derived cell models have been developed for drug metabolism and hepatotoxicity screening to circumvent these drawbacks. Hepatoma cell lines offer important advantages, availability, an unlimited life span and a stable phenotype, thus rendering them suitable models for such studies. However, currently available human hepatoma cell lines are not a good alternative to cultured hepatocytes as they show very limited expression for most drug-metabolising enzymes. Other approaches have been developed to generate immortalised hepatic cells with metabolic competence (use of plasmids encoding immortalising genes to transform human hepatocytes, cell lines obtained from transgenic animals, hepatocytomes or hydrid cells). Recombinant models heterologously expressing cytochrome P450 enzymes in hepatoma cells have also been generated, and are widely used in drug metabolism and toxicity evaluations. In recent years, new approaches to up-regulate the expression of drug-biotransformation enzymes in human cell lines (i.e., transfection with the expression vectors encoding key hepatic transcription factors) have also been investigated. This paper reviews the features of liver-derived cell lines, their suitability for drug metabolism and hepatotoxicity studies, and the state-of-the-art strategies pursued to generate metabolically competent hepatic cell lines.