ROS Induced by KillerRed Targeting Mitochondria (mtKR) Enhances Apoptosis Caused by Radiation via Cyt c/Caspase-3 Pathway
ROS Induced by KillerRed Targeting Mitochondria (mtKR) Enhances Apoptosis Caused by Radiation via Cyt c/Caspase-3 Pathway
复制标题
KillerRed 靶向线粒体 (mtKR) 诱导的 ROS 通过 Cyt c/Caspase-3 途径增强辐射引起的细胞凋亡
DOI：
10.1155/2019/4528616
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发表时间：
2019-01-01
影响因子：
--
通讯作者：
Wang, Zhicheng
中科院分区：
文献类型：
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作者：
Li, Xin;Fang, Fang;Wang, Zhicheng
在放射治疗过程中，活性氧(ROS)诱导的细胞凋亡是放射治疗的主要机制之一。基于KillerRed可以在不同的细胞亚结构中诱导ROS爆发，我们假设KillerRed靶向线粒体(MtKR)可以诱导ROS通过辐射促进细胞凋亡。本研究成功构建了空载体、mtKR和mtmCherry载体，并在COS-7和HeLa细胞中检测到线粒体的定位。用可见光和X射线照射HeLa细胞后，分别检测细胞内ROS水平、线粒体膜电位(Δψm)、三磷酸腺苷(ATPase)活性、三磷酸腺苷(ATPase)含量、细胞凋亡率以及基因和蛋白的表达。结果表明，光照后线粒体ROS水平显著升高，外加辐射后，线粒体电压依赖性阴离子通道1蛋白表达增加，Na+-K+和Ca~(2+)-Δψ~(2+)-ATP酶活性、ATP含量和ATPm显著降低。此外，细胞凋亡率显著增加，即细胞色素c(Cytc)、caspase-9和caspase-3mRNA表达增加，Cytc蛋白从线粒体释放到细胞质中，caspase-9和caspase-3被激活。这些结果表明，mtKR可通过Cytc/caspase-3途径增加辐射诱导的ROS生成，增强线粒体功能障碍，促进细胞凋亡。
During radiotherapy, reactive oxygen species- (ROS-) induced apoptosis is one of the main mechanism of radiation. Based on KillerRed which can induce ROS burst in different cell substructures, here we hypothesized that KillerRed targeting mitochondria (mtKR) could induce ROS to enhance apoptosis by radiation. In this study, empty vector, mtKR, and mtmCherry plasmids were successfully constructed, and mitochondrial localization were detected in COS-7 and HeLa cells. After HeLa cells were transfected and irradiated by visible light and X-rays, ROS levels, mitochondrial membrane potential (Δψm), ATPase activities, adenosine triphosphate (ATP) content, apoptosis, and the expressions of mRNA and protein were measured, respectively. Data demonstrated that the ROS levels significantly increased after light exposure, and adding extra radiation, voltage-dependent anion channel 1 (VDAC1) protein increased in the mitochondria, while Na+-K+ and Ca2+-Mg2+ ATPase activities, ATP content, and Δψm significantly reduced. Additionally, the cell apoptotic rates dramatically increased, which referred to the increase of cytochrome c (Cyt c), caspase-9, and caspase-3 mRNA expressions, and Cyt c protein was released from the mitochondria into the cytoplasm; caspase-9 and -3 were activated. These results indicated that mtKR can increase the production of ROS, enhance mitochondrial dysfunction, and strengthen apoptosis by radiation via Cyt c/caspase-3 pathway.