Pyoluteorin induces cell cycle arrest and apoptosis in human triple-negative breast cancer cells MDA-MB-231
Pyoluteorin induces cell cycle arrest and apoptosis in human triple-negative breast cancer cells MDA-MB-231
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幽门螺杆菌素诱导人三阴性乳腺癌细胞 MDA-MB-231 细胞周期停滞和细胞凋亡
DOI：
10.1111/jphp.13262
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发表时间：
2020-04-04
影响因子：
3.3
通讯作者：
Shen, Yun-Heng
中科院分区：
文献类型：
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作者：
Ding, Ting;Yang, Luo-Jie;Shen, Yun-Heng
目的筛选从土壤真菌黑曲霉中分离得到的6种次生代谢物的细胞毒活性。重要的是，通过体外培养的人乳腺癌MDA-MB-231细胞，探讨其诱导人三阴性乳腺癌MDA-MB-231细胞凋亡的机制。用流式细胞仪检测细胞周期、细胞凋亡率和细胞内活性氧(ROS)的产生。激光扫描共聚焦显微镜检测细胞内ROS生成和线粒体膜电位。结果表明，该药对人乳腺癌细胞株MCF-10A具有明显的选择性杀伤作用(IC50=0.97mM)，而对人乳腺上皮细胞株MCF-10A的杀伤作用较弱。结果表明，该药物可使MDA-MB-231细胞周期停滞于G(2)/M期，并诱导细胞发生凋亡。进一步的实验证明，其诱导细胞凋亡的作用与线粒体膜电位降低、ROS积聚和细胞凋亡相关蛋白表达改变有关。结论该药通过将细胞周期阻滞在G(2)/M期，并通过线粒体途径诱导caspase-3依赖的细胞凋亡，从而对MDA-MB-231细胞具有较强的增殖抑制作用，有望成为治疗三阴性乳腺癌药物的先导化合物。
Objectives To screen the cytotoxic activity of six secondary metabolites isolated from soil fungus Aspergillus niger. Importantly, to investigate the mechanism that pyoluteorin induced human triple-negative breast cancer MDA-MB-231 cells apoptosis in vitro.Methods The cell viability assay was tested with CTG assay. Cell cycle, apoptosis and intracellular reactive oxygen species (ROS) production assay were tested with flow cytometry. Additionally, intracellular ROS production assay and mitochondrial membrane potential assay were determined with laser scanning confocal microscopy. The expression of apoptosis-related proteins was determined with Western blot.Key findings Pyoluteorin displayed significantly selective cytotoxicity against human triple-negative breast cancer MDA-MB-231 cells (IC50 = 0.97 mu m) with low toxicity against human breast epithelial cell MCF-10A. It was found that pyoluteorin could arrest MDA-MB-231 cells cycle at G(2)/M phase and induce cell apoptosis. Further experiments demonstrated that the apoptosis-inducing effect of pyoluteorin was related to reduction of mitochondrial membrane potential, accumulation of ROS and change of apoptosis-related protein expressions.Conclusion Our studies revealed that pyoluteorin had potent proliferation inhibition against MDA-MB-231 cells through arresting cell cycle at G(2)/M phase and inducing caspase-3-dependent apoptosis by mitochondrial pathway, implying that pyoluteorin may be a potential lead compound for drug discovery of human triple-negative breast cancer.