双酚芴暴露于MCF-7乳腺癌细胞可引起细胞全基因组表达谱改变

Changes of Whole Genome Expression Profile of MCF-7 Breast Cancer Cells Exposed to Fluorene-9-Bisphenol

目的:研究内分泌干扰物双酚芴对MCF-7乳腺癌细胞的毒性效应。方法:用不同浓度(10^-5~10^-12 mol/L)的双酚芴分别暴露MCF-7细胞,通过细胞增殖实验(MTS法)研究双酚芴对细胞存活率的影响;采用DCFH-DA荧光染色法测定细胞活性氧水平;利用人类全基因组表达谱芯片分析双酚芴对细胞转录组的影响。结果:细胞增殖实验显示,10^-5 mol/L双酚芴作用于MCF-7细胞,与对照组相比可以显著降低细胞的存活率,其他浓度组无显著差异;细胞活性氧检测结果显示,10^-7~10^-10 mol/L双酚芴组与对照组相比均可以显著提高细胞活性氧水平;全基因组表达谱分析显示,各浓度的双酚芴均能引起细胞全基因组mRNA表达谱发生改变,并将差异表达基因富集到相应的细胞信号通路。结论:双酚芴作用于MCF-7细胞,在特定浓度下可以对细胞存活率、细胞内活性氧水平及细胞全基因组mRNA表达谱产生影响,细胞信号通路分析为进一步验证双酚芴对体外细胞的毒性机制提供了参考依据。

Objective:To study the cytotoxic effect of endocrine disruptor fluorene-9-bisphenol(BHPF)on MCF-7 breast cancer cells.Methods:MCF-7 cells were exposed to different concentrations(10-5~10-12 mol/L)of BHPF.The effect of BHPF on cell viability was studied by cell proliferation assay(MTS).The level of reactive oxygen species was measured by DCFH-DA fluorescence staining.The changes of BHPF on cell transcriptome mRNA was analyzed by whole genome expression microarray.Results:Cell proliferation assay showed that 10-5 mol/L group could significantly reduce the viability of MCF-7 cells compared with the control group,and there was no significant difference in other concentration groups compared with the control group.The detection of the ROS showed that 10-7~10-10 mol/L concentration groups compared with the control group could significantly improve the level of ROS.The whole genome microarray analysis showed that the mRNA expression patterns were changed,and the differentially expressed genes were enriched in the corresponding cell signaling pathways.Conclusion:The BHPF can affect the cell viability,influent the level of intracellular ROS and change the mRNA expression patterns of MCF-7 cells at specific concentrations.The analysis of cell signal pathway provides a reference basis for further verifying the toxicity mechanism of BHPF on cells in vitro.