依布硒诱导多发性骨髓瘤细胞凋亡及其机制的研究

Ebselen induces apoptosis in human multiple myeloma cell line U226

目的探讨依布硒对多发性骨髓瘤细胞增殖、凋亡的影响及其机制。方法应用CCK-8法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,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。

Objective To investigate the effect of ebselen on the proliferation and apoptosis in multiple myeloma （MM） cells and its probably mechanism. Methods Cell viability of human MM cell line U266 was examined by CCK-8 assay kit after treated by ebselen at 0,5,10,20,25,30,35 and 40 μmol/L for 8, 24, 48 or 72 h. The production of reactive oxygen species （ROS） was measured by Singlet ROS Probe. JC-1 dye was used to assay the mitochondrial membrane potential （ Δφm）. The apoptosis assay was done through Ho- echst staining. The protein levels of Bcl-2, Bax, caspase 9 and cytochrome C were detected by Western blot- ting. 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 was released from the mitochondria to cytosol accompanied by mitochondrial membrane potential Δφm 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.