联合肿瘤坏死因子相关凋亡诱导配体和阻断PI3-K-Akt信号通路影响鼻咽癌细胞生长凋亡的实验研究

目的 观察联合肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis-inducing ligand,TRAIL)和阻断PI3-K-Akt信号通路,对鼻咽癌细胞株CNE-2的影响,并探索其可能的机制.方法 用TRAIL和PI3-K特异性抑制剂LY294002单独和联合处理鼻咽癌细胞株CNE-2,采用四甲基偶氮唑蓝法检测细胞存活率,流式细胞仪进行细胞凋亡率的分析,Western blot检测相关蛋白表达及Caspase活化程度.用TRAIL和LY294002单独和联合处理人良性成纤维细胞株MRC-5,检测细胞存活率.结果 TRAIL质量浓度＞1 ng/ml时,联合处理组CNE-2细胞存活率大于TRAIL单独处理组存活率(P值均＜0.05);当TRAIL质量浓度为10 ng/ml和100 ng/ml,联合处理组细胞凋亡率明显大于TRAIL单独处理组细胞凋亡率(t值为7.167和7.206,P值均＜0.05);与TRAIL单独处理组相比,联合处理组出现明显的Caspase-8、Caspase-3、Caspase-9活化增多.两组对比Akt、p-Akt、Bcl-2表达无明显改变;在MRC-5中,对照组、TRAIL单独处理组和联合处理组细胞存活率差异无统计学意义(P＞0.05).结论 联合TRAIL和阻断PI3-K-Akt信号通路对抑制鼻咽癌细胞株CNE-2生长,诱导细胞凋亡具有协同作用,其可能机制是诱导线粒体依赖途径.

Effects of combining tumor necrosis factor related apoptosis-inducing ligand with PI3-K-Akt inhibition on nasopharyngeal carcinoma cell

Objective To study the effects of combinative therapy of tumor necrosis factor related apoptosis-inducing ligand (TRAIL) and PI3-K-Akt inhibitor on the growth and apoptosis of nasopharyngeal carcinoma (NPC) cells and underlying mechanisms.Methods With cell growth assay,flow cytometric analysis and Western blotting,the effects of TRAIL and PI3-K-Akt special inhibitor ( LY294002 ) on cell growth,apoptosis and related proteins expressions in CNE-2 cell lines were studied.Results When concentrate of TRAIL ＞ 1 ng/ml,viability rate of cells in combinative treatment group with TRAIL and LY294002 was higher than that in the single treatment group with TRAIL ( all P ＜ 0.05 ).When concentrate of TRAIL were 10 ng/ml and 100 ng/ml,the combinative treatment induced CNE-2 apoptosis more obviously than single treatments ( t were 7.167 and 7.206,all P ＜ 0.05 ).The combination group showed more cleavage of Caspase-8,Caspase-3,Caspase-9 than single treatment groups.Conclusions Combinative application of TRAIL and PI3-K-Akt pathway inhibitor inhibits the growth of CNE-2 and induces apoptosis.The mitochondrial dependent pathway is implicated for the underlying mechanism.