典型内、外源雌激素非基因组途径诱导的雌激素效应研究

Xenoestrogens (XEs), a kind of endocrine disrupting chemicals (EDCs), can produce estrogen-like effects and exist some latent ecological environment and human health risks inevitably. XEs produce estrogenic effects via genomic and nongenomic pathways. Nongenomic pathways are mediated by G protein-coupled estrogen receptor (GPER), epidermal growth factor receptor (EGFR), membrane estrogen receptor (mERs) and receptor tyrosine kinases (RTKs), which can explain a series of special phenomena induced by XEs, such as low-dose, low-affinity, nonlinear, fast signal transduction and integration effects and so on. In order to fill the deficiency of the current nongenomic pathways, this project studied various XEs and used five cells included MCF-7, T47-D, COS-7, SK-BR-3 and MDA-MB-231 as the experimental material with estrogen sensitivity but different receptor expression levels. To begin with, we obtained the low-dose and nonlinear effects induced by XEs in different cells. And then, we studied the participation levels of different receptors that mediated nongenomic pathways estrogenic effects by the use of antagonists and gene interference technology. At last, we analyzed different estrogenic effects that induced through nongenomic pathways mediated by various XEs and clarified the mechanism of action as well. Implementation of this project is capable of more systemic understanding on the low-dose and nonlinear estrogenic effects caused by XEs, and will be sufficient and convictive in elucidating the distinct fashion in which XEs inducing the nongenomic pathways of estrogenic effects mediated by different receptors, and can provide basic mechanisms information of the estrogenic effects.

外源雌激素（XEs）是内分泌干扰物中的一大类可以产生类雌激素效应的外源荷尔蒙，具有潜在的环境生态及人体健康风险。XEs通过基因组途径和非基因组途径产生雌激素效应。非基因组途径由GPER、EGFR、mERs、RTKs介导，并可以解释XEs诱导产生的低剂量、低亲和力、非线性、快速信号传导、整合效应等一系列特殊现象，但其研究并不深入、系统。为弥补当前非基因组途径研究的不足，本项目选取多种XEs为研究对象，以雌激素敏感且受体表达水平不同的MCF-7等五种细胞为实验材料，获得XEs在不同细胞中诱导的低剂量、非线性效应；通过施加拮抗剂及使用基因干扰技术，对比研究不同受体介导非基因组途径雌激素效应的参与程度；分析XEs通过非基因组途径诱导的雌激素效应并阐明其作用机理。本项目的实施能对多种XEs的低剂量、非线性雌激素效应及不同受体介导的非基因组途径形成系统的认识，为雌激素效应产生机理提供理论指导。

外源雌激素（XEs）是内分泌干扰物中的一大类可以产生类雌激素效应的外源荷尔蒙，具有潜在的环境生态及人体健康风险。XEs通过基因组途径和非基因组途径产生雌激素效应。本项目以多种典型XEs（E2、EE2、BPA、4-CP、DBP、Nar和Qu）为研究对象，采用4种雌激素敏感且受体表达不同细胞系为体外模型（MCF-7、SKBR3、MDA-MB-231和HepG2）研究XEs诱导的雌激素效应及其介导机制。（1）项目系统分析了不同XEs单一暴露下的低剂量、非线性效应，在此基础上，通过检测细胞迁移、侵袭、周期、凋亡、活性氧（ROS）生成和氧化应激酶活性等相关指标，分析了XEs复合暴露对不同体外模型的生物学效应。（2）采用添加不同雌激素受体（ERs）拮抗剂（TAM、G36和Broussoflavonol B）的方式，通过蛋白免疫印迹（WB）和实时荧光定量PCR（RT-qPCR）分析细胞内多种ERs的表达水平及其相对比例变化，并分析其下游信号转导通路，阐明XEs介导的雌激素效应机制。（3）突出研究了典型XEs（E2和BPA）通过基因组和非基因组Crosstalk作用对药物雌激素TAM用于乳腺癌治疗的内分泌抗性贡献。本项目的实施对多种XEs的低剂量、非线性雌激素效应及不同受体介导的非基因组途径形成系统的认识，为其雌激素效应产生机理及健康风险评估提供数据支撑和理论指导。