In vivo bioluminescence imaging to evaluate estrogenic activities of endocrine disrupters
In vivo bioluminescence imaging to evaluate estrogenic activities of endocrine disrupters
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DOI：
10.1016/j.ab.2005.02.032
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发表时间：
2005-05-15
影响因子：
2.9
通讯作者：
Nicolas, JC
中科院分区：
文献类型：
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作者：
Pillon, A;Servant, N;Nicolas, JC
报告基因技术被广泛用于测量激素类似物的活性，体外生物发光试验已经允许快速筛选许多化学物质，以确定新的激素激动剂或拮抗剂，或检测环境中内分泌干扰物的存在。已经建立了稳定的生物发光细胞系，它们提供了可重复的剂量响应曲线和准确测定各种化学物质的体外效率。然而，在体内，这些分子可以被代谢，与蛋白质结合，或储存在脂肪中，因此可以显示出不同于在体外观察到的效率。体内试验，如子宫营养生物试验，需要大量的动物牺牲，反应不仅依赖于所有雌激素作用，还暗示其他因素。为了更快的检测，避免使用大量的动物，我们开发了由稳定的生物发光细胞组成的全体内生物传感器植入裸鼠体内。选择MCF-7生物发光细胞系是因为它们在缺乏雌激素的情况下增殖较低，因此异种移植物的大小可以稳定数周。荧光素酶基因表达用冷却电荷耦合器件相机无创监测。通过定量分析，Lis比较了不同雌激素化合物（雌二醇、雌酮）和内分泌干扰物（乙炔雌二醇、染料木素、辛基酚）在体外和体内的作用。和2.4 '-二氯二苯二氯乙烯)在同一细胞系中，并跟随激素作用，作为时间的函数作用于活的动物。使用了不同的给药方案，大多数产品观察到良好的相关性。然而，我们发现口服乙炔雌二醇是最有效的化学物质。(c) 2005爱思唯尔公司版权所有。
Reporter gene technology is widely used to measure activity of hormone analogs, and bioluminescent in vitro assays have allowed rapid screening of numerous chemicals either to identify new agonists or antagonists of hormones or to detect the presence of endocrine disrupters in the environment. Stable bioluminescent cell lines have been established and they provide reproducible dose-response curves and accurate determination of in vitro efficiencies of various chemicals. In vivo, however, these molecules can be metabolized, bound by proteins, or stored in fats and thus could display efficiencies different from those observed in vitro. In vivo assays, Such as the uterotrophic bioassay, require numerous sacrificed animals, and responses not only are dependent oil all estrogenic action but also imply other factors. For a faster assay and to avoid the use of numerous animals, we developed all in vivo biosensor constituted of stable bioluminescent cells implanted in nude mice. MCF-7 bioluminescent cell lines were chosen since their proliferation is low in the absence of estrogen and the xenograft size can thus be stable for several weeks. Luciferase gene expression was monitored noninvasively with a cooled charge-coupled device camera. Quantitative analysis allowed Lis to compare in vitro and in vivo actions of different estrogenic compounds (estradiol, estrone) and endocrine disruptors (ethynylestradiol, genistein, octylphenol. and 2.4 '-dichlorodiphenyldichloroethylene) in the same cell lines and to follow hormone action oil a living animal as a function of time. Different administration protocols have been used and good correlation was observed for most products. However, we found that ethynylestradiol was the most efficient chemical when orally administered. (c) 2005 Elsevier Inc. All rights reserved.