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目的探讨佛手挥发油对MDA-MB-435人乳腺癌细胞增殖的影响。方法采用MTT法测定细胞活力;倒置显微镜、荧光显微镜和扫描电镜观察细胞的凋亡与坏死的形态学变化;彗星电泳检测细胞DNA的损伤程度;流式细胞术分析细胞周期的变化。结果佛手挥发油对MDA-MB-435癌细胞的增殖有明显的抑制作用,且呈剂量依赖性。不同浓度的佛手挥发油处理MDA-MB-435细胞6h后,低、中浓度实验组(31.25,62.5和125μg.mL-1)细胞体积缩小、核固缩,染色质凝集或片段化,呈现典型的凋亡特征;而高浓度实验组(250μg.mL-1)细胞膜裂解,但核形态完整,呈明显的坏死特征。此外,彗星电泳结果显示,低、中浓度实验组彗星头小而彗尾窄长,表明细胞发生了凋亡;高浓度实验组彗星头大而彗尾长且宽,表明细胞已坏死。进一步的流式细胞结果显示,佛手挥发油能将MDA-MB-435癌细胞周期阻滞在S期和G2/M期。结论佛手挥发油具有抑制MDA-MB-435癌细胞增殖的作用,低、中浓度的佛手挥发油诱导细胞凋亡且将细胞周期阻滞在S期和G2/M期,而高浓度的佛手挥发油引起细胞坏死。
Objective To investigate the effect of volatile oil from bergamot on proliferation of MDA-MB-435 human breast cancer cells. Methods Cell viability was measured by MTT method. Morphological changes of apoptosis and necrosis were observed by inverted microscope, fluorescence microscope and scanning electron microscope. DNA damage was detected by comet assay and cell cycle analysis by flow cytometry. Results Bergamot essential oil significantly inhibited the proliferation of MDA-MB-435 cancer cells in a dose-dependent manner. The cells in low and medium concentration groups (31.25, 62.5 and 125μg.mL-1) were reduced in volume, nuclear pyknosis, chromatin condensation or fragmentation after treated with different concentrations of Bergamot essential oil for 6h. However, in the high concentration group (250μg.mL-1), the cell membrane was lysed, but the nuclear morphology was intact with obvious necrosis characteristics. In addition, the result of cometary electrophoresis showed that the comet head of the low and middle concentration experimental groups was small and the tail length was long and narrow, indicating that the cells were apoptotic. The comet head of the high concentration experimental group was large and the tail was long and wide, indicating that the cells were necrotic. Further flow cytometry results showed that bergamot volatile oil could block the cell cycle of MDA-MB-435 in S phase and G2 / M phase. Conclusions The volatile oil of bergamot can inhibit the proliferation of MDA-MB-435 cells. Low and medium concentrations of bergamot essential oil can induce cell apoptosis and block cell cycle in S phase and G2 / M phase. High concentrations of bergamot essential oil cause cells Necrosis.