双酚A类同系物的雌激素效应及对MCF-7细胞的毒性

文育, 雷炳莉, 康佳, 张凯琼, 许洁

doi:10.3969/j.issn.1007-2861.2014.04.021

上海大学学报(自然科学版) >

2015 , Vol. 21 >Issue 4: 515 - 524

DOI: https://doi.org/10.3969/j.issn.1007-2861.2014.04.021

环境与化学工程

双酚A类同系物的雌激素效应及对MCF-7细胞的毒性

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(上海大学环境与化学工程学院环境污染与健康研究所, 上海200444)

收稿日期: 2013-12-18

网络出版日期: 2015-08-31

基金资助

高校青年骨干教师国内访问学者计划资助项目(60C11113001); 长江学者和创新团队发展计划资助项目(IRT13078)

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Estrogen activity and toxicity effects of bisphenol A analogues on MCF-7 cells

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(Institute of Environmental Pollution and Health, School of Environmental and Chemical Engineering,Shanghai University, Shanghai 200444, China)

Received date: 2013-12-18

Online published: 2015-08-31

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摘要

双酚A(bisphenol A, BPA)及其类似物作为聚碳酸酯的主要合成原料, 一直是环境污染的重要问题, 其环境分布及人体暴露是当今科学研究的热点. 为评估环境中BPA 类物质的毒性效应, 采用MTT 比色法检测了7 种BPA 类同系物对人雌激素受体(estrogen receptor,ER)缺失乳腺癌细胞MCF-7(ER–) 增殖活性的影响. 通过检测细胞乳酸脱氢酶(lactate dehydrogenase, LDH)露出率评估了目标物对MCF-7 细胞膜的损伤, 结果表明, 在0.01～1.00 μmol/L 浓度范围内, 受试化合物对MCF-7 细胞的毒性较小, LDH 少量露出; 而在10 和100 μmol/L 浓度下, 细胞增殖受到显著抑制, 细胞膜通透性显著改变, LDH大量露出. 同时采用ER 基因双杂交酵母实验检验了这几种BPA 类同系物的雌激素效应大小, 通过Logistic 模型对曲线进行了拟合, 并以模型估算的半数透导活性(EC50) 评估了7 种BPA 类同系物的雌激素效应活性, 其大小依次为TDP>BPAF>4Cl-BPA>BPC>BPA>BPE>BPAP. 这说明其雌激素效应机制可能是通过和ER 结合而产生的.

关键词： MCF-7 细胞; 雌激素效应; 双酚A 类同系物; 细胞毒性

本文引用格式

文育, 雷炳莉, 康佳, 张凯琼, 许洁 . 双酚A类同系物的雌激素效应及对MCF-7细胞的毒性[J]. 上海大学学报(自然科学版), 2015 , 21(4) : 515 -524 . DOI: 10.3969/j.issn.1007-2861.2014.04.021

Abstract

Bisphenol A (BPA) and its analogues as main raw materials for synthesis of polycarbonate are important issue of environmental pollution. Most of the current studies focus on PBA analogues’ environmental distribution and human exposure. To evaluate the cytotoxic effects of BPA analogues, MTT assay was used to assess the proliferation toxicity of 7 kinds of BPA analogues on MCF-7 (ER–), when ER is estrogen receptor. At the same time, releases of lactate dehydrogenase (LDH) escaping into cell culture medium were detected to evaluate the damage of cell membrane. The results showed that in the concentration ranges of 0.01～1.00 μmol/L, the toxic effects of tested compounds on MCF-7 cells were small and only a small amount of LDH were released. At concentrations of 10 and 100 μmol/L, cell proliferation was significantly inhibited. Permeability of cell membranewas greatly changed, and lots of LDH were released. In addition, ER gene two-hybrid yeast bioassay was used to evaluate the estrogen effects of BPA analogues. All the dose-response relationships were described by logistic function. The estrogenic activity expressed by EC50 were TDP>BPAF>4Cl-BPA>BPC>BPA>BPE>BPAP. The results shows that the
estrogenic effect mechanism is due to the binding of that target compounds and ER.

Key words： bisphenol A (BPA) analogues; cell toxicity; estrogenic effect; MCF-7 cells

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