Camalexin Induces Apoptosis via the ROS-ER Stress-Mitochondrial Apoptosis Pathway in AML Cells
Camalexin Induces Apoptosis via the ROS-ER Stress-Mitochondrial Apoptosis Pathway in AML Cells
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Camalexin 通过 ROS-ER 应激-线粒体凋亡途径诱导 AML 细胞凋亡
DOI：
10.1155/2018/7426950
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发表时间：
2018-01-01
影响因子：
--
通讯作者：
Yu, Rui
中科院分区：
文献类型：
--
作者：
Yang, Yang;Wang, Gang;Yu, Rui
Camalexin 是一种植物抗毒素，在暴露于环境压力和植物病原体时会在各种十字花科植物中积累。研究表明，camalexin 具有有效的抗肿瘤特性，但其潜在机制仍不清楚。在本研究中，我们评估了camalexin对人类白血病细胞和正常多形核细胞的影响。 CCK-8测定用于测定camalexin处理后的细胞活力。通过流式细胞术测量细胞凋亡、细胞内活性氧（ROS）水平和线粒体膜电位（MMP）损失。测定SOD、过氧化氢酶活性和GSH/GSSG比值。通过Western blot检测ER应激和细胞凋亡信号通路。使用异种移植小鼠来验证camalexin的体内作用。我们的结果表明，camalexin 抑制白血病细胞的活力，但不抑制正常多形核细胞的活力。此外，camalexin 通过线粒体途径以半胱天冬酶依赖性方式诱导细胞凋亡。我们还观察到内质网应激位于camalexin诱导的细胞凋亡的上游。此外，camalexin处理后ROS水平、SOD活性、CAT活性和GSSG水平显着升高，而GSH水平降低。此外，ROS的产生对于camalexin诱导的ER应激和细胞凋亡至关重要。最后，camalexin 的给药抑制了异种移植肿瘤的生长，且没有明显的毒性。综上所述，这项研究表明camalexin通过ROS-ER应激-线粒体凋亡途径对白血病细胞发挥抗肿瘤作用。
Camalexin is a phytoalexin that accumulates in various cruciferous plants upon exposure to environmental stress and plant pathogens. It was shown that camalexin has potent antitumor properties, but its underlying mechanisms are still elusive. In the present study, we evaluated the effects of camalexin on human leukemic cells and normal polymorph nuclear cells. CCK-8 assay was used to determine cell viability after camalexin treatment. Apoptosis, intracellular reactive oxygen species (ROS) levels, and loss of mitochondrial membrane potential (MMP) were measured by flow cytometry. The activity of SOD, catalase, and ratio of GSH/GSSG were assayed. ER stress and apoptotic signaling pathway was examined by Western blot. Xenograft mice were used to verify the effect of camalexin in vivo. Our results indicated that camalexin inhibited viability of leukemic but not normal polymorph nuclear cells. Furthermore, camalexin induces apoptosis via the mitochondrial pathway in a caspase-dependent manner. We also observed ER stress is located upstream of apoptosis induced by camalexin. Besides, ROS levels, SOD activity, CAT activity, and GSSG levels were significantly enhanced while the GSH level was decreased after treatment of camalexin. In addition, the generation of ROS is critical for the ER stress and apoptosis induced by camalexin. Finally, administration of camalexin suppresses xenograft tumor graft growth without obvious toxicity. Taken together, this study indicates that camalexin exerts antitumor effects against leukemia cells via the ROS-ER stress-mitochondrial apoptosis pathway.