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使用水污染物评估三种亚致死细胞毒性试验对人肝癌细胞系HepG2的敏感性。

Evaluation of the sensitivity of three sublethal cytotoxicity assays in human HepG2 cell line using water contaminants.

作者信息

Jondeau Adeline, Dahbi Laurence, Bani-Estivals Marie-Hélène, Chagnon Marie-Christine

机构信息

ENSBANA, UMR (1234) Toxicologie Alimentaire, 1 Esplanade Erasme, F-21000 Dijon, France.

出版信息

Toxicology. 2006 Sep 21;226(2-3):218-28. doi: 10.1016/j.tox.2006.07.007. Epub 2006 Jul 10.

Abstract

The in vitro toxicological index IC50 (the millimolar concentration of compound which inhibits response assay by 50% compared to the solvent control) of 11 water contaminants (acrylamide, atrazine, B[a]P, BPA, 2,4-DAT, 17-alphaEE, H(2)O(2), 4-OP, sodium bromate, sodium chlorate, sodium nitrate) was evaluated on the human hepatoma (HepG2) cells using three short-term bioassays related to their morbidity status [radiometric RNA synthesis assay (RNA), luminometric ATP assay (ATP), fluorometric Alamar blue assay (AB)]. Among all substances, we were not able to determine atrazine IC50 value whatever the test used. Furthermore, B[a]P was not cytotoxic in the ATP and AB assays. Statistical analysis revealed a correlation between the IC50 values obtained in the three assays. Except with 4-OP, RNA assay was always inhibited at lower concentrations than those required in the other assays, suggesting that this assay is a very sensitive indicator of the presence of toxic compounds. ATP and AB assays responded to a similar pattern. Due to its higher sensitivity and its reliability, RNA synthesis assay using HepG2 cell line provides the most suitable tool for the screening of water contaminants.

摘要

使用与人类肝癌(HepG2)细胞发病状态相关的三种短期生物测定法[放射性RNA合成测定法(RNA)、发光ATP测定法(ATP)、荧光Alamar蓝测定法(AB)],评估了11种水中污染物(丙烯酰胺、阿特拉津、苯并[a]芘、双酚A、2,4-二氨基甲苯、17-α乙炔雌二醇、过氧化氢、4-辛基酚、溴酸钠、氯酸钠、硝酸钠)的体外毒理学指标IC50(与溶剂对照相比,抑制反应测定50%的化合物的毫摩尔浓度)。在所有物质中,无论使用何种测试,我们都无法确定阿特拉津的IC50值。此外,苯并[a]芘在ATP和AB测定中无细胞毒性。统计分析表明,三种测定法获得的IC50值之间存在相关性。除4-辛基酚外,RNA测定法在比其他测定法所需浓度更低时总是受到抑制,这表明该测定法是有毒化合物存在的非常敏感的指标。ATP和AB测定法呈现相似模式。由于其更高的灵敏度和可靠性,使用HepG2细胞系的RNA合成测定法为筛选水中污染物提供了最合适的工具。

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