Department of Obstetrics and Gynecology, University Hospital Freiburg, Freiburg, Germany.
Reprod Toxicol. 2010 Aug;30(1):89-93. doi: 10.1016/j.reprotox.2010.02.002. Epub 2010 Feb 19.
The human endometrium is a fertility-determining factor. Its receptivity during the implantation window may be altered by chemicals. Since human embryo implantation is unique chemical risk assessment cannot be based solely on animal studies. We established a tissue-specific in vitro test based on human endometrial adenocarcinoma (Ishikawa) cells. Progesterone receptor (PR) was selected as primary target gene for estrogenic effects. Changes of mRNA levels were investigated by reverse transcription quantitative real-time PCR. Sigmoidal dose-response curves for up-regulation of PR mRNA and EC(50) values were established for 17beta-estradiol, diethylstilbestrol and the weak xenoestrogen bisphenol A. Nonylphenol also had a clear PR mRNA up-regulating effect. Several other chemicals were characterized as negative compounds. Among them was methoxyacetic acid which may produce false positive results in reporter gene assays. Up-regulation of PR protein by 17beta-estradiol, diethylstilbestrol, bisphenol A and nonylphenol was confirmed by Western Blotting.
人类子宫内膜是决定生育能力的因素。其在着床窗口的接受能力可能会被化学物质改变。由于人类胚胎着床是独特的，因此不能仅基于动物研究进行化学风险评估。我们基于人子宫内膜腺癌（Ishikawa）细胞建立了一种组织特异性的体外测试。孕激素受体（PR）被选为雌激素效应的主要靶基因。通过反转录定量实时 PCR 研究 mRNA 水平的变化。建立了 17β-雌二醇、己烯雌酚和弱的外源性雌激素双酚 A 上调 PR mRNA 的 S 型剂量反应曲线和 EC50 值。壬基酚也具有明显的 PR mRNA 上调作用。其他几种化学物质被表征为阴性化合物。其中包括甲氧乙酸，它可能会在报告基因检测中产生假阳性结果。Western Blotting 证实了 17β-雌二醇、己烯雌酚、双酚 A 和壬基酚对 PR 蛋白的上调作用。
