超氧阴离子和细胞外信号调节激酶在金雀异黄素抑制人乳腺癌细胞浸润中作用的研究
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中国博士后基金(2011M501254,2012T50484);江苏省博士后基金


Genistein suppresses MDA-MB-231 breast cancer cells invasion via decreasing superoxide anion-dependent ERK activation
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    摘要:

    目的:研究金雀异黄素对乳腺癌细胞(MDA-MB-231)浸润能力的影响及其可能机制-方法:不同浓度的金雀异黄素(25~100 μmol/L) 孵育MDA-MB-231细胞24 h,用Transwell assay检测细胞浸润能力;用免疫印迹法比较细胞内磷酸化和总细胞外信号调节激酶(ERK)蛋白含量的改变;用免疫荧光实验检测细胞内磷酸化ERK的分布;用DHE染色及荧光显微镜检测细胞内超氧阴离子水平-结果:金雀异黄素处理使细胞浸润能力降低;同时,金雀异黄素的处理引起细胞内磷酸化ERK量明显降低,且磷酸化ERK在核内含量明显下降;ERK抑制剂U0126作用于细胞亦能明显降低细胞的浸润能力;金雀异黄素处理还能使细胞内超氧阴离子含量明显下降,超氧阴离子清除剂TEMPOL不仅明显抑制磷酸化ERK含量,同时细胞浸润能力也明显降低-结论:金雀异黄素可能通过抑制乳腺癌细胞的超氧阴离子水平和ERK活性,进而抑制MDA-MB-231乳腺癌细胞的浸润能力-

    Abstract:

    Objective:To investigate the effect of genistein on the invasion of breast cancer cells (MDA-MB-231) and its possible mechanism. Methods:MDA-MB-231 cells were co-cultured with genistein at various concentrations (25 to 100 μmol/L) for 24 h. Cell invasion ability was studied by Transwell invasion assay. Phosphorylated ERK and total ERK proteins were quantified by Western blot analysis. Distribution of cellular phosphorylated ERK was detected by fluorescence microscope. Intracellular superoxide anion production was stained by DHE staining and detected by fluorescence microscope. Results:After treated with genistein for 24 h,the invasion ability of MDA-MB-231 cells was significantly reduced in a dose-dependent manner compared to the control cells. Consistently,ERK activity and p-ERK location in the nucleus also decreased after genistein treatment. Furthermore,ERK inhibitor U0126 could inhibit cell invasion similarly as genistein. In addition,superoxide anion production was suppressed when treated with genistein,and superoxide anion scavenger TEMPOL could not only suppress ERK phosphorylation,but also inhibit cell invasion. Conclusion:Genistein can inhibit invasive ability of MDA-MB-231 cells,probably through inhibiting the superoxide anion/ERK pathway.

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金建明,杜 军.超氧阴离子和细胞外信号调节激酶在金雀异黄素抑制人乳腺癌细胞浸润中作用的研究[J].南京医科大学学报(自然科学版),2012,(10):1351-1355

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  • 收稿日期:2012-04-27
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