依布硒诱导多发性骨髓瘤细胞凋亡及其机制的研究.docVIP

依布硒诱导多发性骨髓瘤细胞凋亡及其机制的研究 张 亮，杜 佳，周立为，程 燚，王 东，谢家印 （400042重庆，第三军医大学大坪医院野战外科研究所肿瘤中心） [摘要] 目的 探讨依布硒对多发性骨髓瘤细胞增殖、凋亡的影响及机制。方法 应用CCK8观察不同浓度依布硒作用于人多发性骨髓瘤细胞株U266细胞8、24、48、72 h后细胞活力及增殖情况，同时应用Hoechst染色检测细胞凋亡，JC-1检测细胞线粒体膜电位，ROS探针观察依布硒对细胞内活性氧的影响，以及Western blot检测Bcl-2、Bax、细胞色素C等凋亡相关蛋白及Caspase9激活情况。结果 依布硒能够显著抑制多发性骨髓瘤细胞增殖，并诱导细胞凋亡，40μmol/L依布硒使凋亡率达（80.8±3.9）%；同时依布硒能升高细胞内ROS水平，降低线粒体膜电位；此外依布硒可诱导细胞色素C从线粒体向胞浆释放但并不激活Caspase9。 结论 依布硒可诱导多发性骨髓瘤细胞发生非Caspase依赖的细胞凋亡，其机制可能与ROS水平升高有关。 [关键词] 依布硒；多发性骨髓瘤；凋亡；活性氧 [中图法分类号] [文献标志码] A Ebselen induces apoptosis in human multiple myeloma cell lines and its mechanism Zhang Liang, Du Jia, Zhou Liwei, Cheng Yi, Wang Dong, Xie Jiayin (Cancer center, Daping Hospital and Institute of Surgery Research, Third Military Medical University, Chongqing，4000042, China) [Abstract]Objective To investigate the effect of Ebselen on cell proliferation and apoptosis in multiple myeloma (MM) cells and its probably mechanism. Methods Cell viability of human MM cell line U266 was examined by CCK8 assay kit. The production of reactive oxygen species (ROS) was measured by Singlet ROS Probe. JC-1 was used to assay the mitochondrial membrane potential (△Ψm). The apoptosis assay was done through Hoechst staining. The protein levels of Bcl-2, Bax, caspase 9 and cytochrome c were detected by Western blot. Results Ebselen was found to significantly decrease cell viability and increase apoptosis accompanied by enhancing production of ROS in U266 cells. Further studies revealed cytochrome c release from the mitochondria to cytosol clines accompanied by mitochondrial membrane potential △Ψm decrease, while caspase 9 was not observed to be activated. Conclusion Ebselen may enhance the production of endogenous ROS, and induce MM cell apoptosis via mitochondria-mediated apoptotic pathway. [Key words] Ebselen Multiple Myeloma Apoptosis ROS Ebselen induce apoptosis in human multiple myeloma cell lines and its mechanism Zhang Liang, Du Jia, Zhou Liwei, Cheng Y