BMS-345541通过抑制IκB激酶/核因子-κB信号通路诱导人脑胶质瘤细胞凋亡

BMS-345541 induces apoptosis of glioma cells through inhibition of IκB kinase/nuclear factor-κB pathway

目的 观察IκB激酶（IKK）高度选择性抑制剂BMS-345541对胶质瘤细胞凋亡的影响及其机制.方法 不同浓度BMS-345541作用于人脑胶质瘤细胞株U87MG后,采用Annexin V-FITC/PI双标记法检测细胞凋亡,荧光素酶核因子-κB（NF-κB）报告基因法检测NF-κB活性,免疫印迹法检测bcl-2、Caspase-3和NF-κB胞核定位.结果 终质量浓度1、10、20 μmol/L BMS-345541作用U87MG细胞24 h后,细胞凋亡率分别为（18.2±2.2）%、（49.3±3.6）%、（73.3±8.9）%,与对照组比较差异有统计学意义（P＜0.01）.BMS-345541处理U87MG细胞后,bcl-2蛋白表达减少、Caspase-3被激活;NF-κB转录启动子的活性下降（66.2±5.1）%（P＜0.01）,NF-κB p65亚单位核移位被抑制.结论 BMS-345541可通过抑制IKK/NF-κB信号通路诱导人胶质瘤细胞凋亡.

Objective To investigate the effect of BMS-345541, a novel compound with a highly selective IκB kinase （IKK） inhibitory activity, on apoptosis of glioma cells and the possible mechanisms.Methods Human glioma cells U87MG were treated with different concentrations of BMS-345541. Cell apoptosis was investigated by Annexin V/PI staining assay. B cell lymphoma gene-2 （bcl-2） and Caspase-3 was detected by Western blotting. Nuclear factor-κB （NF-κB） activity was analyzed by NF-κB luciferase reporter gene assay and nuclear translocation of NF-κB was analyzed by Western blotting. Results After incubation with 1,10 and 20 μmol/L BMS-345541 ,apoptosis rates of U87MG cells were （18. 2 ±2. 2）% ,（49. 3 ± 3.6 ） % and （ 73.3 ± 8.9 ） %, respectively （ compared with the control, P 〈 0. 01 ）. BMS-345541 induced the activation of Caspase-3 and decreased expression of bcl-2. Moreover, NF-κB activity was decreased by（ 66. 2 ± 5.1 ）% （ P 〈 0. 01 ） and nuclear translocation of NF-κB p65 subunit was inhibited.Conclusion BMS-345541 induced apoptosis of glioma cells involving inhibition of IKK/NF-κB signaling pathway.