The MCF-7cell was used as a research model to evaluate the toxicological effects and estrogenic effects of dibutyl phthalate (DBP) and bisphenol A (BPA). Four analysis methods, including MTT assay, DCFH-DA fluorescent staining, flow cytometry, and real-time fluorescent quantitative PCR (RT-QPCR), were applied to evaluate the effects of DBP and BPA on cell activity (CA), reactive oxygen species (ROS) formation, cell cycle distribution and cell apoptosis, and mRNA transcription levels of three estrogen receptors (e.g.,ERα,ERβ and GPR30), respectively. CA represented an inverted U-shape, in which both low dose (10?8mol/L) and high dose (10?4mol/L) exposure to DBP and BPA inhibited cell proliferation but stimulated cell proliferation ranged from 10?7mol/L to 10?5mol/L. The MCF-7cells had the highest cell proliferation rate when exposure concentrations were 10?6mol/L and 10?7mol/L for DBP and BPA, respectively. And the interaction of DBP and BPA was additive action under low-concentration but showed antagonistic effect under high-concentration. On one hand, co-exposure to low-doses DBP and BPA could obviously induce cell proliferation with cell cycle arrest in S phase and mRNA transcription induction of ERα and GPR30; on the other hand, co-exposure to high-doses of DBP and BPA significantly induced cell apoptosis as they induced ROS generation, blocked cell cycle in G0/G1phase, and inhibited the mRNA transcription of ERα. Therefore, the results will provide experimental data and theoretical direction to comprehensively evaluate the potential health risk of co-exposure of alkyl phenols and phthalic acid esters.%以乳腺癌细胞系MCF-7为雌激素效应和毒性效应评价模型,分别采用MTT法检测邻苯二甲酸二丁酯(DBP)和双酚A(BPA)对细胞活力的影响,采用DCFH-DA荧光染色法测定细胞活性氧水平,采用流式细胞仪检测细胞周期和凋亡,采用RT-QPCR检测细胞中ERα、ERβ和GPR30mRNA的转录水平.结果表明:DBP和BPA对MCF-7细胞活力呈现"倒U"型剂量-效应关系,即低浓度(10?8mol/L)和高浓度(10?4mol/L)抑制细胞增殖,在10?7~10?5mol/L浓度范围内刺激细胞增殖,并分别在浓度为10?6和10?7mol/L时达到最高增殖率;DBP和BPA的联合暴露中,低浓度联合暴露呈现加和效应,而高浓度则呈现拮抗效应;低浓度联合暴露促进细胞增殖,其产生原因与细胞周期由G0/G1期向S期推进、ERα和GPR30转录诱导相关;而高浓度联合暴露则显著抑制细胞增殖,其产生原因与诱导活性氧自由基(ROS)生成、细胞周期G0/G1期阻滞、ERα 表达抑制相关.研究结果可为环境介质中烷基酚类和酞酸酯类物质共存下的潜在健康风险评估和预防提供基础数据和理论指导.
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