Expression of cytochromes P-450 2E1, 3A4 and 1A1/1A2 in growing and confluent human HepG2 hepatoma cells - Effect of ethanol
Expression of cytochromes P-450 2E1, 3A4 and 1A1/1A2 in growing and confluent human HepG2 hepatoma cells - Effect of ethanol
复制标题
DOI：
10.1016/s0887-2333(99)00007-7
复制
发表时间：
1999-06-01
影响因子：
3.2
通讯作者：
Richert, L
中科院分区：
文献类型：
--
作者：
Alexandre, E;David, P;Richert, L
在培养的人肝癌 HepG2 细胞中，细胞色素 (CYP) 1A 相关的 7-乙氧基试卤灵 O-脱乙基酶 (EROD)、CYP 3A 相关的苄氧基试卤灵 O-脱苄酶 (BROD) 和 CYP 2E1 相关的对硝基苯酚羟化酶 (PNPH) 在培养过程中减少。汇合时细胞内酶活性为接种后24小时细胞内酶活性的35-60%。同样，CYP 3A 和 CYP 2E1 蛋白在生长 (G) 细胞中的浓度高于汇合 (C) HepG2 细胞中的浓度。在 HepG2 细胞 G 和 C 中均未检测到 CYP 1A1/1A2 蛋白，但可通过 3-甲基胆蒽 (3-MC) 处理强烈诱导。乙醇 (EtOH) 显示可增加 HepG2 细胞中 CYP 2E1 和 CYP 3A 蛋白以及 CYP 1A1/1A2-、CYP 2E1- 和 CYP 3A 相关的混合功能氧化酶活性 (MFO)，正如之前针对人肝细胞原代培养物的报道。仅在培养开始时，在生长的 HepG2 细胞中观察到这些效应，证明了 HepG2 细胞的生长阶段对其对 EtOH 处理的反应的影响。据我们所知，这是第一份关于 EtOH 在 HepG2 细胞中增加 CYP 蛋白和相关 MFO 的报告，它表明生长的 HepG2 细胞提供了一个有用的体外模型系统，可用于研究 EtOH 对人类 CYP 的调节。 (C) 1999 Elsevier Science Ltd. 保留所有权利。
In cultured human hepatoma HepG2 cells, cytochrome (CYP) 1A-associated 7-ethoxyresorufin-O-deethylase (EROD), CYP 3A-associated benzyloxyresorufin O-debenzylase (BROD) and CYP 2E1-associated p-nitrophenol-hydroxylase (PNPH) decreased during time in culture. The enzyme activities in cells at confluence were 35-60% of the activities in cells 24 hours after seeding. Similarly, CYP 3A and CYP 2E1 proteins were present at higher concentrations in growing (G) than in confluent (C) HepG2 cells. CYP 1A1/1A2 protein was not detected, neither in G nor in C HepG2 cells but was strongly induced by 3-methylcholanthrene (3-MC) treatment. Ethanol (EtOH) was shown to increase CYP 2E1 and CYP 3A proteins and CYP 1A1/1A2-, CYP 2E1- and CYP 3A-associated mixed-function oxidase activities (MFOs) in HepG2 cells, as has been previously reported for primary cultures of human hepatocytes. These effects were observed only at the beginning of culture, in growing HepG2 cells, demonstrating the influence of the growth stage of HepG2 cells on their response to EtOH treatment. This is, to our knowledge, the first report on increases in CYP proteins and associated MFOs by EtOH in HepG2 cells, it suggests that growing HepG2 cells provide a useful in vitro model system in which to study the regulation of human CYPs by EtOH. (C) 1999 Elsevier Science Ltd. All rights reserved reserved.