Assaying estrogenicity by quantitating the expression levels of endogenous estrogen-regulated genes
Assaying estrogenicity by quantitating the expression levels of endogenous estrogen-regulated genes
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DOI：
10.2307/3454380
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发表时间：
2000-05-01
影响因子：
10.4
通讯作者：
Leffers, H
中科院分区：
文献类型：
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作者：
Jorgensen, M;Vendelbo, B;Leffers, H
科学证据表明，人类和野生动物物种可能会因接触与内分泌系统相互作用的环境化学品而经历不利的健康后果。需要可靠的短期检测来识别干扰荷尔蒙的化学物质。在这项研究中，我们证明了一种化学物质的雌激素活性可以通过检测内源性雌激素调节的“标记基因”在人乳腺癌MCF-7细胞中的诱导或抑制来评估。我们包括四个标记基因-PS2，转化生长因子β3，单胺氧化酶A，和α1-抗糜蛋白酶。我们评估了17β-雌二醇(E-2)、己烯雌酚、α-玉米赤霉醇、壬基酚、染料木素、甲氧氯胺、硫丹、邻苯二甲酸二丁酯、双酚A、邻苯二甲酸二丁酯、4-羟基他莫昔芬和ICI 182.780的雌激素活性。所有四个标记基因都对三种高效雌激素(E-2、己烯雌酚和α-玉米赤霉醇)有强烈的反应，而其他化学物质的效力比E-2低10(3)到10(6)倍。受试化学物质的相对效力存在一些标记基因相关的差异。转化生长因子β3对三种高效雌激素的敏感性相同，而与其他三个标记基因一起检测时，对α-玉米赤霉醇的敏感性比对E-2和己烯雌酚的敏感性低约10倍。当用PS2和转化生长因子β3测定时，壬基酚的效价与金雀异黄素相同，而用单胺氧化酶A和α1-抗凝乳糜酶测定时，其效价分别高/低10~100倍。这一结果与其他方法获得的结果一致，表明基于内源基因表达的检测方法可能提供了一种替代其他E-Screen方法的有吸引力的替代方法。
Scientific evidence suggests that humans and wildlife species may experience adverse health consequences from exposure to environmental chemicals that interact with the endocrine system. Reliable short-term assays are needed to identify hormone-disrupting chemicals. In this study we demonstrate that the estrogenic activity of a chemical can be evaluated by assaying induction or repression of endogenous estrogen-regulated "marker genes" in human breast cancer MCF-7 cells. We included four marker genes in the assay-pS2, transforming growth factor beta 3 (TGF beta 3), monoamine oxidase A, and alpha 1-antichymotrypsin-and we evaluated estrogenic activity for 17 beta-estradiol (E-2), diethylstilbestrol, alpha-zearalanol, nonylphenol, genistein, methoxychlor, endosulphan, o,p-DDE, bisphenol A, dibutylphthalate, 4-hydroxy tamoxifen, and ICI 182.780. All four marker genes responded strongly to the three high-potency estrogens (E-2, diethylstilbestrol, and alpha-zearalanol), whereas the potency of the other chemicals was 10(3)- to 10(6)-fold lower than that of E-2. There were some marker gene-dependent differences in the relative potencies of the tested chemicals. TGF beta 3 was equally sensitive to the three high-potency estrogens, whereas the sensitivig to alpha-zearalanol was approximately 10-fold lower than the sensitivity to E-2 and diethylstilbestrol when assayed with the other three marker genes. The potency of nonylphenol was equal to that of genistein when assayed with pS2 and TGF beta 3, but 10- to 100-fold higher/lower with monoamine oxidase A and alpha 1-antichymotrypsin, respectively. The results are in agreement with results obtained by other methods and suggest that an assay based on endogenous gene expression may offer an attractive alternative to other E-SCREEN methods.