摘要
目的探讨荜茇明碱对人乳腺癌MDA-MB-231细胞的放射增敏作用及其可能机制。方法体外培养MDA-MB-231细胞,取对数生长期细胞进行实验。将细胞分为对照组、X射线照射组、荜茇明碱处理组、荜茇明碱联合X射线照射组。克隆集落形成法分析荜茇明碱对MDA-MB-231细胞的放射增敏作用,按多靶单击模型拟合细胞存活曲线,确定存活分数(SF)、平均致死剂量(D0)、准阈剂量(Dq),计算放射增敏比(SER)。用流式细胞仪(FCM)检测细胞凋亡;用Western blot观察凋亡相关蛋白的表达;DCFH-DA探针结合FCM分析细胞内活性氧水平的变化。结果克隆形成试验显示,与单纯X射线照射组细胞相比,荜茇明碱联合X射线照射组细胞克隆形成能力下降,SF2、D0及Dq均下降(P均<0.05),其放射增敏比(SERD0)为1.22及1.29。与单纯X射线照射组相比,荜茇明碱联合X射线照射组细胞凋亡明显增加。与对照组和单独处理组相比,荜茇明碱与X射线联合处理组使Bcl-2表达明显减少,Bax的表达明显增加。荜茇明碱提高了X射线照射引起的细胞内活性氧的水平。结论低浓度的荜茇明碱增加了MDA-MB-231细胞的放射敏感性,可能与其调控凋亡相关蛋白表达并且升高细胞内活性氧水平,从而增加X射线诱导的细胞凋亡有关。
[Abstract] Objective To investigate the radio-sensitivity enhancement effect of piplartine on human breast cancer MDA-MB-231 cells line and to discuss the possible mechanism of radio-sensitivity enhancement effect. Methods Human breast cancer cell line MDA- MB- 231 was cultured in vitro and the cells in logarithmic growth phase were selected for experiments. MDA- MB- 231 cells were divided into four groups： control, X-ray expose, piplartine, and piplartine combined with X-rays. The clonogenic assay was performed to determine the radio-sensitivity enhancement effect of piplartine on MDA-MB-231 cells. The cell survival curve was fitted by single-hit multi-target model and then the survival fraction （SF）, average lethal dose （Do）, quasi-threshold dose （Dq） and sensitizing enhance rate （SER） were calculated. Cell apoptosis was analyzed by flow cytometry （FCM）. Western blot assay was employed to measure the expression of apoptosis- related protein （Bel- 2 and Bax proteins） after treatment with piplartine and/or X- ray radiation. The intracellular reactive oxygen species （ROS） level was detected by FCM with the probe of DCFH-DA. Results The cloning formation capacity was decreased in the group of piplartine plus radiation, which showed that the values of SF2, Do, Dq were significantly lower than those of radiation alone group（P 〈 0.05）, and the sensitizing enhancement rate （SER） of Do was 1.22 and 1.29. The cell apoptosis rate was increased by the combination treatment of piplartine and radiation. Piplartine increased the radiation- induced intracellular levels of
出处
《介入放射学杂志》
CSCD
北大核心
2014年第2期147-152,共6页
Journal of Interventional Radiology
基金
江苏省医学创新团队与领军人才资助项目(LJ201123)
