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首页> 外文期刊>Journal of biomedical optics >Single-cell analysis of dihydroartemisinin-induced apoptosis through reactive oxygen species-mediated caspase-8 activation and mitocnondrial pathway in ASTC-a-1 cells using fluorescence imaging techniques
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Single-cell analysis of dihydroartemisinin-induced apoptosis through reactive oxygen species-mediated caspase-8 activation and mitocnondrial pathway in ASTC-a-1 cells using fluorescence imaging techniques

机译：通过荧光成像技术对双氢青蒿素通过活性氧介导的caspase-8活化和线粒体途径在ASTC-a-1细胞中诱导的细胞凋亡进行单细胞分析

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Dihydroartemisinin (DHA), a front-line antimalarial herbal compound, has been shown to possess promising anticancer activity with low toxicity. We have previously reported that DHA induced caspase-3-dependent apoptosis in human lung adenocarcinoma cells. However, the cellular target and molecular mechanism of DHA-induced apoptosis is still poorly defined. We use confocal fluorescence microscopy imaging, fluorescence resonance energy transfer, and fluorescence recovery after photobleaching techniques to explore the roles of DHA-elicited reactive oxygen species (ROS) in the DHA-induced Bcl-2 family proteins activation, mitochondrial dysfunction, caspase cascade, and cell death. Cell Counting Kit-8 assay and flow cytometry analysis showed that DHA induced ROS-mediated apoptosis. Confocal imaging analysis in a single living cell and Western blot assay showed that DHA triggered ROS-dependent Bax translocation, mitochondrial membrane depolarization, alteration of mitochondrial morphology, cytochrome c release, caspase-9, caspase-8, and caspase-3 activation, indicating the coexistence of ROS-mediated mitochondrial and death receptor pathway. Collectively, our findings demonstrate for the first time that DHA induces cell apoptosis by triggering ROS-mediated caspase-8/Bid activation and the mitochondrial pathway, which provides some novel insights into the application of DHA as a potential anticancer drug and a new therapeutic strategy by targeting ROS signaling in lung adenocarcinoma therapy in the future.
机译：一线抗疟疾草药化合物双氢青蒿素（DHA）已被证明具有低毒的抗癌活性。我们以前曾报道过DHA诱导人肺腺癌细胞中caspase-3依赖性凋亡。但是，DHA诱导的细胞凋亡的细胞靶标和分子机制仍然不清楚。我们使用共聚焦荧光显微镜成像，荧光共振能量转移和光漂白技术后的荧光回收，以探讨DHA诱导的活性氧（ROS）在DHA诱导的Bcl-2家族蛋白激活，线粒体功能障碍，胱天蛋白酶级联，和细胞死亡。细胞计数试剂盒8检测和流式细胞仪分析表明DHA诱导ROS介导的细胞凋亡。在单个活细胞中进行共聚焦成像分析和Western blot分析表明，DHA触发了ROS依赖性Bax移位，线粒体膜去极化，线粒体形态改变，细胞色素c释放，caspase-9，caspase-8和caspase-3活化，表明ROS介导的线粒体和死亡受体途径的共存。总的来说，我们的发现首次证明DHA通过触发ROS介导的caspase-8 / Bid激活和线粒体途径诱导细胞凋亡，这为DHA作为潜在的抗癌药物的应用和新的治疗策略提供了一些新颖的见解。通过在未来的肺腺癌治疗中靶向ROS信号转导。

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