REGULATION OF CYTOCHROME-P450 EXPRESSION IN A NOVEL LIVER-CELL LINE FROM ZEBRAFISH (BRACHYDANIO-RERIO)
REGULATION OF CYTOCHROME-P450 EXPRESSION IN A NOVEL LIVER-CELL LINE FROM ZEBRAFISH (BRACHYDANIO-RERIO)
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DOI：
10.1006/abbi.1993.1429
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发表时间：
1993-09-01
影响因子：
3.9
通讯作者：
BUHLER, DR
中科院分区：
文献类型：
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作者：
MIRANDA, CL;COLLODI, P;BUHLER, DR
研究了2,3,7,8-四氯二苯并-对二恶英（TCDD）和β-萘黄酮（BNF）在成年斑马鱼（Brachydanio rerio）新肝细胞系中细胞色素P450的表达和诱导作用。对照、BNF或tcdd处理细胞的亚细胞组分在免疫印迹中未显示可检测到的鳟鱼P450组成形式（LMC1、LMC2、LMC3、LMC4和LMC5）的抗体，这表明斑马鱼肝细胞缺乏与鳟鱼组成形式表达的P450密切相关的P450，或者同源P450的浓度太低而无法检测到。然而，暴露于TCDD后，细胞表达了虹鳟鱼P4501A1抗体识别的主要免疫反应性54-kDa蛋白和次要50-kDa蛋白。这些免疫反应蛋白在tcdd处理的细胞的微粒体和线粒体部分中观察到，但在二甲亚砜（DMSO）（对照）或BNF处理的细胞培养中未检测到。TCDD显著提高了该细胞株的乙氧基间苯二酚脱乙基酶（EROD）和7,12-二甲基苯并蒽（DMBA）羟化酶的活性，而BNF没有显著提高。经tcdd处理的肝细胞对α-萘黄酮和抗鳟鱼P4501A1 IgG等诊断抑制剂的EROD活性比DMBA羟化酶活性更敏感。经tcdd处理的细胞将DMBA转化为各种代谢物，其中一种是假定的近似致癌物DMBA-3,4-二醇。这些结果表明，TCDD，而不是BNF，在培养的斑马鱼肝细胞中诱导一种或可能两种形式的P450免疫化学和功能上与鳟鱼P4501A1相关。
The expression and induction of cytochrome P450 by 2,3,7,8-tetrachlordibenzo-p-dioxin (TCDD) and β-naphthoflavone (BNF) in a new liver cell line from adult zebrafish (Brachydanio rerio) were studied. Subcellular fractions from control, BNF- or TCDD-treated cells did not show detectable bands in immunoblots probed with antibodies to the constitutive forms of trout P450 (LMC1, LMC2, LMC3, LMC4, and LMC5), suggesting that either zebrafish liver cells lack P450s closely related to those constitutively expressed in trout or that the concentrations of the orthologous P450s were too low to be detected. However, upon exposure to TCDD, the cells expressed a major immunoreactive 54-kDa protein and a minor 50-kDa protein recognized by antibodies to rainbow trout P4501A1. These immunoreactive proteins were observed in microsomal and mitochondrial fractions of TCDD-treated cells but were not detected in cell cultures treated with dimethyl sulfoxide (DMSO) (vehicle control) or BNF. The activities of ethoxyresorufinO-deethylase (EROD) and 7,12-dimethylbenzanthracene (DMBA) hydroxylase were markedly increased by TCDD but not by BNF in this cell line. EROD activity was more sensitive than DMBA hydroxylase activity of TCDD-treated liver cells to diagnostic inhibitors such as α-naphthoflavone and anti-trout P4501A1 IgG. The TCDD-treated cells converted DMBA to various metabolites, one of which is the putative proximate carcinogen, DMBA-3,4-diol. These results suggest that TCDD, but not BNF, induces one or possibly two forms of P450 immunochemically and functionally related to trout P4501A1, in cultured zebrafish liver cells.