Development of a human lymphoblastoid cell line constitutively expressing human CYP1B1 cDNA: Substrate specificity with model substrates and promutagens
Development of a human lymphoblastoid cell line constitutively expressing human CYP1B1 cDNA: Substrate specificity with model substrates and promutagens
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DOI：
10.1093/mutage/12.2.83
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发表时间：
1997-03-01
期刊：
影响因子：
2.7
通讯作者：
Sutter, TR
中科院分区：
文献类型：
--
作者：
Crespi, CL;Penman, BW;Sutter, TR
在染色体外载体上稳定表达人细胞色素P4501B1 (CYP1B1) cDNA并共表达NADPH细胞色素P450氧化还原酶（OR）的AHH-1 TK+/-细胞衍生物被命名为h1B1/OR，表达CYP1B1 cDNA的细胞制备的微粒体表现出7-乙氧基-再间酚去乙基酶（EROD）、7-乙氧基-4-三氟甲基香豆素去乙基酶（EFCD）水平升高。苯并(a)芘羟化酶（BPH）、丁呋喃醇1′-羟化酶、睾酮羟化酶活性和可光谱定量的细胞色素P450，含有CYP1B1的微粒体不含可检测到的香豆素7-羟化酶、对硝基酚羟化酶、月桂酸羟化酶、(S)-甲苯托因4′-羟化酶或双氯芬酸4′-羟化酶活性。对于BPH和EFCD，底物代谢率排序为CYP1A1>CYP1B1>CYP1A2，对于EROD，底物代谢率排序为CYP1A1>CYP1A2>CYP1B1。对于EROD和EFCD， CYP1B1的表观K-m值更接近于CYP1A1而不是CYP1A2。为了开始表征CYP1B1的促原激活能力，我们在h1B1/OR细胞中检测了所选化学物质的致突变性；与对照细胞相比，表达CYP1B1的细胞对苯并(a)芘、环五（c,d）芘、4-（甲基亚硝胺）-1-（3-吡啶基）-1-丁酮和黄曲霉毒素B-1 （AFB）的致突变性敏感性增加，在该系统中表达的CYP1B1似乎对激活AFB特别有效。
An AHH-1 TK+/- cell derivative was developed that stably expresses human cytochrome P4501B1 (CYP1B1) cDNA in an extrachromosomal vector which confers resistance to I-histidinol and co-expresses NADPH cytochrome P450 oxidoreductase (OR), The CYP1B1-expressing cell line was designated h1B1/OR, Microsomes prepared from CYP1B1 cDNA expressing cells exhibit elevated levels of 7-ethoxy-resorufin deethylase (EROD), 7-ethoxy-4-trifluoromethyl-coumarin deethylase (EFCD), benzo(a)pyrene hydroxylase (BPH), bufuralol 1'-hydroxylase, testosterone hydroxylase activities and spectrally quantifiable cytochrome P450, CYP1B1-containing microsomes did not contain detectable coumarin 7-hydroxylase, p-nitrophenol hydroxylase, lauric acid hydroxylase, (S)-mephenytoin 4'-hydroxylase or diclofenac 4'-hydroxylase activities, Kinetic parameters for selected substrates were compared among CYP1B1 and the two additional members of the CYP1 family, CYP1A1 and CYP1A2, For BPH and EFCD, the rank order of rates of substrate metabolism were CYP1A1>CYP1B1>CYP1A2, For EROD, the rank order of substrate metabolism was CYP1A1>CYP1A2>CYP1B1. For both EROD and EFCD the apparent K-m values for CYP1B1 were more similar to CYP1A1 than to CYP1A2, In order to begin to characterize the promutagen activating ability of CYP1B1, the mutagenicity of selected chemicals was examined in h1B1/OR cells; there was increased sensitivity (CYP1B1-expressing relative to control cells) to the mutagenicity of benzo(a)pyrene, cyclopenta(c,d)pyrene, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and aflatoxin B-1 (AFB), CYP1B1, expressed in this system, appears to be particularly efficient at activating AFB.