典型环境内分泌干扰物对MCF-7细胞的增殖毒性及初步机理探讨

发布时间：2018-06-23 03:31

本文选题：雌激素类化合物 + 沉积物有机提取物　；参考：《上海大学》2013年硕士论文

【摘要】：雌激素类化合物是典型的环境内分泌干扰物（environmental endocrinedisrupting chemicals, EDCs），在低剂量下就具有较强的生殖及发育毒性，成为环境毒理学领域的关注热点之一。其中雌二醇（E2）、戊酸雌二醇（EV）、己烯雌酚（DES）、双酚A（BPA）、DEHP及其代谢产物MEHP被认为是典型雌激素类化合物，在水环境中广泛存在，受到人们的关注，但关于它们的联合毒性效应及作用机制仍需加强研究。本文运用经典的人类乳腺癌细胞MCF-7增殖实验，采用CCK8法测试了典型雌激素类化合物对MCF-7细胞增殖的单一毒性及不同实验设计下的联合毒性效应；并从雌激素受体和细胞内活性氧水平变化方面探讨了雌激素干扰物对MCF-7细胞增殖毒性作用途径；同时进行了黄浦江和苏州河沉积物有机提取物对MCF-7细胞增殖的复合毒性效应及对细胞内ROS水平的影响，主要结果如下：（1）E2、EV、DES、BPA促进了人类乳腺癌细胞MCF-7的增殖，而DEHP、MEHP对MCF-7细胞增殖没有影响；析因实验和等浓度配比两种实验设计研究DES和EV的二元混合物对MCF-7细胞的增殖效应均表现为加和效应或接近加和效应。（2）目标化合物E2、DES、BPA对MCF-7细胞的ER受体蛋白有一个明显的表达，说明它们可能是通过雌激素受体（ER）调节机制对MCF-7细胞增殖产生促进作用的。此外E2、EV和BPA都可调节MCF-7细胞内ROS水平，，他们在诱导氧化应激的同时，又发挥着抗氧化性机制，使细胞内氧化还原水平处于动态平衡中；DES和DEHP对ROS水平影响不明显；DES、BPA、DEHP与EV联合作用没有改变这三种化合物单独作用时对细胞内ROS水平的影响。（3）黄浦江和苏州河沉积物的有机提取物对MCF-7细胞增殖的毒性测试结果显示，除个别点外，大部分样点均表现出不同程度的MCF-7细胞增殖的毒性效应，且呈剂量-效应依赖和时间-效应依赖关系。并对MCF-7细胞内ROS水平产生一定的影响。说明黄浦江、苏州河沉积物受到了有机污染物的污染，并且部分河段污染较严重，但具体是受到哪类污染物污染需要进一步进行化学分析确认。
[Abstract]:Estrogen compounds are typical environmental endocrine disruptors (environmental endocrinedisrupting chemicals,), which have strong reproductive and developmental toxicity at low doses, and have become one of the focuses in the field of environmental toxicology. Among them, estradiol (E2), estradiol valerate (EV), diethylstilbestrol (des), bisphenol A (BPA) DEHP and its metabolite MEHP are considered to be typical estrogen compounds, which are widely existed in water environment and have attracted people's attention. However, the study of their joint toxicity and its mechanism need to be strengthened. In this paper, the single toxicity of typical estrogen compounds to MCF-7 cell proliferation and the combined toxicity under different experimental designs were tested by CCK8 method using the classical human breast cancer cell MCF-7 proliferation test. The toxic effects of estrogen disruptors on MCF-7 cell proliferation were also discussed from the aspects of estrogen receptor and intracellular reactive oxygen species (Ros) levels. At the same time, the compound toxic effects of organic extracts from sediments from Huangpu River and Suzhou River on the proliferation of MCF-7 cells and their effects on intracellular Ros levels were studied. The main results were as follows: (1) E2EVV DESPA promoted the proliferation of human breast cancer cell line MCF-7. The proliferation of MCF-7 cells was not affected by DEHP- MEHP. The effects of des and EV binary mixtures on the proliferation of MCF-7 cells were studied by factorial experiment and equal concentration ratio. (2) the ER response of the target compound E2DSE BPA to MCF-7 cells was either additive effect or near additive effect. (2) the ER response of the target compound E2DSE BPA to MCF-7 cells was studied. Body protein has an obvious expression. These results suggest that they may promote the proliferation of MCF-7 cells through estrogen receptor (ER) regulation. In addition, both E2EV and BPA could regulate the Ros level in MCF-7 cells. They not only induced oxidative stress, but also played an antioxidant mechanism, so that the intracellular redox level was in a dynamic balance. The effects of des and DEHP on Ros levels were not obvious. The combined action of DESPA-DEHP and EV did not change the effect of these three compounds on intracellular Ros levels. (3) the toxicity test of organic extracts from sediments of Huangpu River and Suzhou River to MCF-7 cell proliferation showed that, except for a few points, Most of the samples showed toxic effects of MCF-7 cell proliferation to varying degrees, and showed dose-effect and time-effect dependence. The Ros level in MCF-7 cells was also affected to some extent. The results show that the sediments of Huangpu River and Suzhou River are polluted by organic pollutants, and some of them are polluted seriously, but it is necessary to make chemical analysis to confirm which kinds of pollutants are polluted.
【学位授予单位】：上海大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114;X503.1

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