Establishment of the transformants expressing human cytochrome P450 subtypes in HepG2, and their applications on drug metabolism and toxicology.
Establishment of the transformants expressing human cytochrome P450 subtypes in HepG2, and their applications on drug metabolism and toxicology.
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HepG2表达人细胞色素P450亚型转化子的建立及其在药物代谢和毒理学中的应用。
DOI：
10.1016/s0887-2333(01)00011-x
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
S. Asahi
中科院分区：
文献类型：
--
作者：
S. Yoshitomi;K. Ikemoto;J. Takahashi;H. Miki;M. Namba;S. Asahi
建立了在人肝细胞系 HepG2 中稳定表达一系列人细胞色素 P450 (CYP) 亚型的转化子。这些转化子被命名为Hepc/1A1.4、Hepc/1A2.9、Hepc/2A6L.14、Hepc/2B6.68、Hepc/2C8.46、Hepc/2C9.1、Hepc/2C19.12、Hepc/2D6.39、Hepc/2E1.3-8和Hepc/3A4.2-30，其稳定表达 分别为人类 CYP1A1、CYP1A2、CYP2A6、CYP2B6、CYP2C8、CYP2C9、CYP2C19、CYP2D6、CYP2E1 和 CYP3A4。通过酶活性测定和逆转录聚合酶链式反应（RT-PCR）程序证实了转化体中CYP亚型的表达。表达的 CYP 亚型对其特定底物的表观 Km 值与人肝微粒体的表观 Km 值接近。除了 CYP 活性外，这些转化体还保留了葡萄糖醛酸和硫酸盐结合活性。此外，CYP2C9、CYP2D6和CYP3A4的活性被它们的特异性抑制剂抑制。通过表达 CYP 的转化体分析对乙酰氨基酚 (APAP)、环磷酰胺 (CPA) 和苯并[a]蒽 (BA) 的细胞毒性。细胞毒性取决于CYP亚型的表达，并以剂量依赖性方式增加。这些结果显示了转化体中表达的特定CYP亚型对APAP、CPA和BA的代谢激活，并证明了这些转化体对于人肝脏的体外代谢和毒理学研究的有用性。
Transformants with stable expression of a series of human cytochrome P450 (CYP) subtypes in the human hepatic cell line, HepG2, were established. These transformants are designated Hepc/1A1.4, Hepc/1A2.9, Hepc/2A6L.14, Hepc/2B6.68, Hepc/2C8.46, Hepc/2C9.1, Hepc/2C19.12, Hepc/2D6.39, Hepc/2E1.3-8 and Hepc/3A4.2-30, which stably expressed human CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4, respectively. The expression of the CYP subtypes in the transformants was confirmed by both determination of enzyme activities and the reverse transcriptase polymerase chain reaction (RT-PCR) procedure. The apparent Kmvalues of the expressed CYP subtypes for their specific substrates were close to those of human liver microsomes. In addition to their CYP activities, these transformants retained glucuronide- and sulfate-conjugating activities. Furthermore, the activities of CYP2C9, CYP2D6 and CYP3A4 were inhibited by their specific inhibitors. The cytotoxicity of acetaminophen (APAP), cyclophosphamide (CPA) and benz[a]anthracene (BA) were analyzed by CYP-expressing transformants. The cytotoxicity depended on the expression of CYP subtypes and increased in a dose-dependent manner. These results show the metabolic activation of APAP, CPA and BA by the specific CYP subtypes expressed in the transformants and demonstrate the usefulness of these transformants for in vitro metabolic and toxicological studies in human liver.
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影响因子：
3.6
作者：
Chen, Q;Cederbaum, AI
通讯作者：
Cederbaum, AI
影响因子：
3.9
作者：
PATTEN, CJ;ISHIZAKI, H;YANG, CS
通讯作者：
YANG, CS
影响因子：
3.9
作者：
RAUCY, JL;LASKER, JM;BLACK, M
通讯作者：
BLACK, M
DOI：
10.1016/s0021-9258(17)38834-8
发表时间：
1985-10
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. Moore;H. Thor;G. Moore;Sid Nelsons;P. Moldéus;S. Orrenius
通讯作者：
M. Moore;H. Thor;G. Moore;Sid Nelsons;P. Moldéus;S. Orrenius
基因工程细胞色素 P450 毒性作用的分子研究。
影响因子：
5.9
作者：
Doehmer,J;Buters,JT;Luch,A;Soballa,V;Baird,WM;Morisson,H;Stegeman,JJ;Townsend,AJ;Greenlee,WF;Glatt,HR;Seidel,A;Jacob,J;Greim,H
通讯作者：
Greim,H