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雀梅藤Sageretia thea(Osbeck)Johnst又名雀梅刺、对角刺、碎米子等,为鼠李科雀梅藤属植物,主要分布地区包括云南、安徽、江苏等[1-2]。根、茎、叶均可入药,其性甘、淡、平,具有降气化痰、祛风利湿等功效[3]。民间常用雀梅藤药材煎汤或浸酒内服治疗乳腺肿瘤、淋巴结肿大和水肿等疾病,具有较好效果[4]。现代药理研究表明,雀梅藤具有抗结直肠癌、抗炎、抗氧化、抗病毒、抗菌和肝保护等活性[5-6],但尚未见雀梅藤对乳腺癌等其他肿瘤影响的报道。该研究通过多种体外实验观察雀梅藤对乳腺癌细胞增殖及凋亡的影响,并初步探索其作用机制,以证实民间应用该药材治疗乳腺肿瘤的合理性,为进一步的药物开发奠定基础。
Induction of apoptosis in triple-negative breast cancer cells by petroleum ether extract of Sageretia thea
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摘要:
目的 探讨雀梅藤石油醚提取物对乳腺癌细胞增殖和凋亡的影响。 方法 石油醚提取物孵育乳腺癌细胞不同时间后,CCK8法检测细胞活力,平板克隆形成法检测细胞增殖,DAPI染色法观察细胞核形态变化,免疫荧光检测线粒体膜电位和活性氧变化,流式细胞术检测细胞周期和凋亡,CCK8法观察对人正常血管内皮细胞毒性。 结果 雀梅藤石油醚提取物孵育BT549和MDA-MB-231人三阴性乳腺癌细胞株24 h后的IC50分别为45.40 μg/ml和12.23 μg/ml,时间和剂量依赖地抑制乳腺癌细胞活力和克隆形成,诱导细胞凋亡,阻滞细胞周期于G1/S期,降低线粒体膜电位,升高活性氧;对人正常血管内皮细胞无明显毒性作用。 结论 雀梅藤石油醚提取物可能通过升高活性氧,引起线粒体膜电位坍塌,激活线粒体通路诱导细胞凋亡,从而抑制乳腺癌细胞生长,支持其民间抗乳腺肿瘤的应用。 Abstract:Objective To investigate the effect of the petroleum ether extract of Sageretia thea on the proliferation and apoptosis of breast cancer cells. Methods After breast cancer cells were incubated with the petroleum ether extract for different times, cell viability was analyzed by CCK8 assay, cell proliferation was detected by plate cloning test, nuclear morphology was observed by DAPI staining, mitochondrial membrane potential(MMP)and reactive oxygen species(ROS)were determined by immunofluorescence, and the cell cycle and apoptosis were detected by flow cytometry. After incubating with the extract for 24 h, the CCK8 assay was used to observe the toxicity to normal human vascular endothelial cells. Results The IC50 of BT549 and MDA-MB-231 breast cancer cell lines treated with the petroleum ether extract for 24 h were 45.40 μg/ml and 12.23 μg/ml, respectively. The extract time and dose dependently inhibited breast cancer cell viability and clonal formation, induced cell apoptosis and cycle arrest in G1/S phase, decreased MMP and increased ROS levels. There was no toxic effect on normal endothelial cells. Conclusion The petroleum ether extract of Sageretia thea may induce apoptosis by increasing ROS to cause MMP collapse, followed by activating mitochondrial pathway, thereby hindering the growth of breast cancer cells. These results could support the application of Sageretia thea to anti-breast tumor in the folk. -
Key words:
- Sageretia thea /
- Breast cancer /
- Proliferation /
- Apoptosis /
- Mitochondrion /
- ROS
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