CAJ | 학술논문

目的探讨雷公藤甲素(TPL)对替莫唑胺(TMZ)杀伤胶质瘤细胞效应的影响及其可能机制.方法采用CCK-8法检测TPL与TMZ对胶质瘤细胞增殖的抑制效应,使用Chou-Talalay法评估TPL和TMZ的联合作用,流式细胞仪检测技术检测TPL和(或)TMZ作用后U87胶质瘤细胞的周期和凋亡的变化,以及应用Western blot技术检测TPL和(或)TMZ作用后U87胶质瘤细胞中IκBα、磷酸化IκBα、XIAP及Cleaved PARP表达的变化.结果 TPL与TMZ作为单药均能够剂量依赖性的抑制胶质瘤细胞增殖;在U87细胞系中,TPL联合TMZ抑制U87细胞达IC50时联合指数(CI)为0.76,表明TPL与TMZ具有协同作用;TPL与TMZ联用处理U87细胞后,细胞凋亡比例显著多于单药用药组(t=14.474,P＜0.01;t=11.244,P＜0.01);TPL与TMZ联用可抑制NF-κB信号转导通路中关键蛋白IκBα的磷酸化以及抗凋亡蛋白XIAP的表达.结论 TPL在体外能够协同增加TMZ杀伤胶质瘤细胞的效应,其机制可能通过抑制NF-κB信号转导通路的活化,进一步诱导胶质瘤细胞凋亡所致.
목적 탐토뢰공등갑소(TPL)대체막서알(TMZ)살상효질류세포효응적영향급기가능궤제.방법 채용CCK-8법검측TPL여TMZ대효질류세포증식적억제효응,사용Chou-Talalay법평고TPL화TMZ적연합작용,류식세포의검측기술검측TPL화(혹)TMZ작용후U87효질류세포적주기화조망적변화,이급응용Western blot기술검측TPL화(혹)TMZ작용후U87효질류세포중IκBα、린산화IκBα、XIAP급Cleaved PARP표체적변화.결과 TPL여TMZ작위단약균능구제량의뢰성적억제효질류세포증식;재U87세포계중,TPL연합TMZ억제U87세포체IC50시연합지수(CI)위0.76,표명TPL여TMZ구유협동작용;TPL여TMZ련용처리U87세포후,세포조망비례현저다우단약용약조(t=14.474,P＜0.01;t=11.244,P＜0.01);TPL여TMZ련용가억제NF-κB신호전도통로중관건단백IκBα적린산화이급항조망단백XIAP적표체.결론 TPL재체외능구협동증가TMZ살상효질류세포적효응,기궤제가능통과억제NF-κB신호전도통로적활화,진일보유도효질류세포조망소치.
Objective This study aimed to explore the effect of triptolide (TPL) on the efficiency of temozolomide (TMZ) in killing glioma cells and the possible mechanism.Methods CCK-8 assay was used to test the growth inhibition effect of TMZ and TPL against the glimoa cell.The combined effects of TPL and TMZ on glioma cells were calculated by Chou-Talalay method.Propidium iodide staining,Annexin Ⅴ-FITC/PI staining,and Western-blot were used to detect the change of cell cycle,apoptosis,and its related protein treated by TPL and/or TMZ in glioma cells.Results TPL and TMZ reduced the growth rate of glioma cell line U87 in a dose-dependent manner.CI values of TPL and TMZ are less than 1,indicating the synergistic effect of the two agents.The apoptosis induced by combined TPL and TMZ was more significant than that by either single agent treatment group in U87 cells.Western blot analysis showed that TPL resulted in the down-regulation of anti-apoptotic protein XIAP and the dephosphorylation of IκBα a key component in NF-κB pathway.Conclusions TPL synergistically acted with TMZ to kill glioma cells.TPL enhanced the TMZ-induced apoptosis in glioma cells.Suppression of NF-κB pathway was the possible mechanism.