Can hepatoma cell lines be redifferentiated to be used in drug metabolism studies?
Can hepatoma cell lines be redifferentiated to be used in drug metabolism studies?
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DOI：
10.1177/026119290403201s11
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发表时间：
2004-06-01
影响因子：
2.7
通讯作者：
Castell, JV
中科院分区：
文献类型：
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作者：
Martínez-Jiménez, CP;Jover, R;Castell, JV
在药物开发中，对代谢、迄今为止涉及的酶以及新化合物潜在的酶抑制或酶诱导特性的了解是一个关键问题。原代培养的肝细胞、细胞色素P450 （CYP）工程细胞和肝癌细胞系目前被用于此目的，但只有原代培养才能产生与体内发现的药物相似的代谢谱，并能对诱导剂产生反应。由于它们的可及性有限，目前正在探索替代人类肝细胞的替代品，包括通过使用不同策略（即SV40 t -大抗原，有条件永生化肝细胞，转染c-myc， cH-ras， N-ras癌基因，转基因动物过度表达生长因子或癌基因和cre-lox重组/切除）实现肝细胞的永生化。然而，所得到的细胞没有一个具有理想的表型特征来取代药物代谢研究中的原代培养物。我们研究了为什么这些分化的人肝癌不表达CYP基因，并发现某些关键转录因子的水平与肝细胞中的水平明显不同。因此可以想象，这些转录因子中的一个（或多个）的重新表达可能导致CYP基因的有效转录。通过对Hep G2细胞进行富集肝脏转录因子的基因工程，然后分析最相关的人类cyp的表达，证明了这一假设的可行性。
Knowledge of metabolism, enzymes so far involved, and potential enzyme-inhibiting or enzyme-inducing properties of new compounds is a key issue in drug development. Primary cultured hepatocytes, cytochrome P450 (CYP)-engineered cells and hepatoma cell lines are currently being used for this purpose, but only primary cultures can produce a metabolic profile of a drug similar to that found in vivo and can respond to inducers. Because of their limited accessibility, alternatives to replace human hepatocytes are currently being explored, including the immortalisation of hepatocytes by using different strategies (i.e. SV40 T-large antigen, conditionally immortalised hepatocytes, transfection with c-myc, cH-ras, N-ras oncogenes, transgenic animals over-expressing growth factors or oncogenes and cre-lox recombination/excision). However, none of the resulting cells has the desirable phenotypic characteristics to replace primary cultures in drug metabolisms studies. We investigated why these differentiated human hepatomas do not express CYP genes and found that the levels of certain key transcription factors clearly differ from those found in hepatocytes. It was then conceivable that re-expression of one (or more) of these transcription factors could lead to an efficient transcription of CYP genes. The feasibility of this hypothesis was demonstrated by genetic engineering of Hep G2 cells with liver-enriched transcription factors followed by the analysis of the expression of the most relevant human CYPs.