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泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究

王筱婧 王东兴 范洁 高越 张海

王筱婧, 王东兴, 范洁, 高越, 张海. 泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究[J]. 药学实践与服务, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012
引用本文: 王筱婧, 王东兴, 范洁, 高越, 张海. 泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究[J]. 药学实践与服务, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012
WANG Xiaojing, WANG Dongxing, FAN Jie, GAO Yue, ZHANG Hai. Inhibitory effect of Euphorbia helioscopia on human triple-negative breast cancer MDA-MB-231 cells and its possible mechanism of apoptosis[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012
Citation: WANG Xiaojing, WANG Dongxing, FAN Jie, GAO Yue, ZHANG Hai. Inhibitory effect of Euphorbia helioscopia on human triple-negative breast cancer MDA-MB-231 cells and its possible mechanism of apoptosis[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012

泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究

doi: 10.3969/j.issn.1006-0111.2017.04.012

Inhibitory effect of Euphorbia helioscopia on human triple-negative breast cancer MDA-MB-231 cells and its possible mechanism of apoptosis

  • 摘要: 目的 研究泽漆对三阴乳腺癌MDA-MB-231细胞的作用及其作用机制。 方法 用MTT法检测细胞活力;用荧光显微镜法测定MDA-MB-231细胞的活性氧(ROS)生成量;用流式细胞仪检测细胞的凋亡率;用TUNEL检测法检测细胞凋亡DNA碎片;用Western blot检测caspase-9、caspase-3、PARP等凋亡相关因子的水平变化。 结果 MTT试验显示泽漆提取物对MDA-MB-231细胞具有显著的抑制作用,但作用可被ROS抑制剂NAC及caspase抑制剂Z-VAD-FMK所消除;荧光显微镜检测显示泽漆提取物能显著提高ROS的生成;流式细胞仪检测显示泽漆提取物处理后,PI染色阳性细胞明显增加,但被NAC减弱。Caspase-9、caspase-3在提取物处理后均转为激活形式,PARP被剪切。TUNEL法显示,提取物处理后细胞凋亡碎片明显增多,而提前加入ROS抑制剂NAC和caspase抑制剂Z-VAD-FMK能使泽漆提取物诱导凋亡的DNA碎片明显减少。 结论 泽漆乙酸乙酯提取物可以有效抑制MDA-MB-231细胞的生长,诱导其凋亡,其作用机制可能与ROS过量生成所致的线粒体损伤途径有关。
  • [1] Campbell RA, Bhat-nakshatri P, Patel NM, et al. Phosphatidylinositol 3-kinase/AKT-mediated activation of estrogen receptor alpha: a new model for anti-estrogen resistance[J]. J Biol Chem, 2001, 276(13): 9817-9824.
    [2] Hseu YC, Lee CC, Chen YC, et al. Antrodia salmonea in submerged culture exhibits antioxidant activities in vitro and protects human erythrocytes and low-density lipoproteins from oxidative modification[J]. Food Chem Toxicol, 2014, 66(4): 150-157.
    [3] Shah SP, Roth A, Goya R, et al. The clonal and mutational evolution spectrum of primary triple-negative breast cancers[J]. Nature, 2012, 486(7403): 395-399.
    [4] Cleator S, Heller W, Coombes RC. Triple-negative breast cancer: therapeutic options[J]. Lancet Oncol, 2007, 8(3): 235-244.
    [5] Wang K, Zhang C, Bao J, et al. Synergistic chemopreventive effects of curcumin and berberine on human breast cancer cells through induction of apoptosis and autophagic cell death[J]. Sci rep, 2016, 6: 26064.
    [6] Jeong HS, Choi HY, Lee ER, et al. Involvement of caspase-9 in autophagy-mediated cell survival pathway[J]. Biochim Biophys Acta, 2011, 1813(1): 80-90.
    [7] Green DR. Apoptotic pathways: paper wraps stone blunts scissors[J]. Cell, 2000, 102(1): 1-4.
    [8] 杨 莉, 陈海霞, 高文远. 泽漆化学成分及药理作用研究进展[J]. 中草药, 2007, 38(10): 1585-1589.
    [9] D'Anneo A, Carlisi D, Lauricella M, et al. Parthenolide generates reactive oxygen species and autophagy in MDA-MB231 cells. A soluble parthenolide analogue inhibits tumour growth and metastasis in a xenograft model of breast cancer[J]. Cell Death Dis, 2013, 4(10): e891.
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    [11] Hseu YC, Lee CC, Chen YC, et al. The anti-tumor activity of Antrodia salmonea in human promyelocytic leukemia (HL-60) cells is mediated via the induction of G(1) cell-cycle arrest and apoptosis in vitro or in vivo[J]. J Ethnopharmacol, 2014, 153(2): 499-510.
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  • 收稿日期:  2017-05-27
  • 修回日期:  2017-07-03

泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究

doi: 10.3969/j.issn.1006-0111.2017.04.012

摘要: 目的 研究泽漆对三阴乳腺癌MDA-MB-231细胞的作用及其作用机制。 方法 用MTT法检测细胞活力;用荧光显微镜法测定MDA-MB-231细胞的活性氧(ROS)生成量;用流式细胞仪检测细胞的凋亡率;用TUNEL检测法检测细胞凋亡DNA碎片;用Western blot检测caspase-9、caspase-3、PARP等凋亡相关因子的水平变化。 结果 MTT试验显示泽漆提取物对MDA-MB-231细胞具有显著的抑制作用,但作用可被ROS抑制剂NAC及caspase抑制剂Z-VAD-FMK所消除;荧光显微镜检测显示泽漆提取物能显著提高ROS的生成;流式细胞仪检测显示泽漆提取物处理后,PI染色阳性细胞明显增加,但被NAC减弱。Caspase-9、caspase-3在提取物处理后均转为激活形式,PARP被剪切。TUNEL法显示,提取物处理后细胞凋亡碎片明显增多,而提前加入ROS抑制剂NAC和caspase抑制剂Z-VAD-FMK能使泽漆提取物诱导凋亡的DNA碎片明显减少。 结论 泽漆乙酸乙酯提取物可以有效抑制MDA-MB-231细胞的生长,诱导其凋亡,其作用机制可能与ROS过量生成所致的线粒体损伤途径有关。

English Abstract

王筱婧, 王东兴, 范洁, 高越, 张海. 泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究[J]. 药学实践与服务, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012
引用本文: 王筱婧, 王东兴, 范洁, 高越, 张海. 泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究[J]. 药学实践与服务, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012
WANG Xiaojing, WANG Dongxing, FAN Jie, GAO Yue, ZHANG Hai. Inhibitory effect of Euphorbia helioscopia on human triple-negative breast cancer MDA-MB-231 cells and its possible mechanism of apoptosis[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012
Citation: WANG Xiaojing, WANG Dongxing, FAN Jie, GAO Yue, ZHANG Hai. Inhibitory effect of Euphorbia helioscopia on human triple-negative breast cancer MDA-MB-231 cells and its possible mechanism of apoptosis[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(4): 337-340,358. doi: 10.3969/j.issn.1006-0111.2017.04.012
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