CAJ | 학술논문

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.
목적 관찰IκB격매(IKK)고도선택성억제제BMS-345541대효질류세포조망적영향급기궤제.방법 불동농도BMS-345541작용우인뇌효질류세포주U87MG후,채용Annexin V-FITC/PI쌍표기법검측세포조망,형광소매핵인자-κB(NF-κB)보고기인법검측NF-κB활성,면역인적법검측bcl-2、Caspase-3화NF-κB포핵정위.결과 종질량농도1、10、20 μmol/L BMS-345541작용U87MG세포24 h후,세포조망솔분별위(18.2±2.2)%、(49.3±3.6)%、(73.3±8.9)%,여대조조비교차이유통계학의의(P＜0.01).BMS-345541처리U87MG세포후,bcl-2단백표체감소、Caspase-3피격활;NF-κB전록계동자적활성하강(66.2±5.1)%(P＜0.01),NF-κB p65아단위핵이위피억제.결론 BMS-345541가통과억제IKK/NF-κB신호통로유도인효질류세포조망.
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.