摘要
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6 h后，低、中浓度实验组(31.25，62.5和125 μg·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(250 μg·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。
Abstract
OBJECTIVE To investigate the effects of fingered citron essential oil on proliferation of MDA-MB-435 cells in vitro. METHODS Cell viability was assayed using MTT method，and the morphological alterations of apoptosis or necrosis in MDA-MB-435 cells were observed by inverted microscopy，fluorescence and scanning electron microscope.Furthermore，DNA damage was measured by comet assay，and cell cycle was measured by flow cytometry. RESULTS Essential oil from fingered citron caused concentration dependent inhibitory effect on the proliferation of MDA-MB-435 cells，with IC50 value of （110±5） μg·mL-1.Moreover，treated with different concentations of fingered citron essential oil for 6 h，the cells in the low and middle concentrations experiment groups(31.25，62.5， 125 μg·mL-1) exhibited the apoptotic changes in morphology such as cell shrinkage，chromatin condensation and fragments.In contrast，the cells in the high concentration experiment group(250 μg·mL-1) showed typical necrotic characteristics including membrane rupture and nuclear integrity.Futhermore， the results from comet assay showed that comet heads were smaller and comet tails were longer in the low or middle concentration experiment groups， than that in high concentration experiment groups，illustrating that cells ocurred apoptosis or necrosis，respectively.In addition，flow cytometry analysis showed that cell cycle was arrested in S and G2/M stages. CONCLUSION Essential oil from fingered citron could inhibit proliferation of MDA-MB-435 cells in a concentration-dependent manner in vitro. Moreover，low and middle concentrations of essential oil from fingered citron could induce apoptosis and the cell cycle was arrested in S and G2/M stages，whereas high concentrations of the compouds resulted in necrosis in MDA-MB-435 cells.
关键词
佛手 /
挥发油 /
细胞凋亡 /
坏死 /
MDA-MB-435癌细胞
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Key words
fingered citron /
essential oil /
apoptosis /
necrosis /
MDA-MB-435 carcinoma cells
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麻艳芳 邵邻相 张均平 邓刚 吕学维 徐玲玲 毕洁琼.
佛手挥发油对MDA-MB-435人乳腺癌细胞体外增殖的影响[J]. 中国药学杂志, 2010, 45(22): 1737-1741
M Yn-fng;SHO Lin-xing;ZHNG Jun-ping;DENG Gng;L Xue-wei;XU Ling-ling;I Jie-qiong.
Effects of Fingered Citron Essential Oil on Proliferation of MDA-MB-435 Cells in Vitro[J]. Chinese Pharmaceutical Journal, 2010, 45(22): 1737-1741
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