氟达拉滨与TRAIL联用诱导肺癌细胞凋亡及其作用机制

Fludarabine combined with TRAIL induced apoptosis in lung cancer A549 cells

目的:研究氟达拉滨(Fludarabine)在低浓度下与TRAIL联合处理是否能够诱导人肺癌A549细胞发生凋亡,并探讨其作用机制。方法:实验设对照组、Fludarabine组、TRAIL组、Fludarabine和TRAIL联合组,CCK-8法检测Fludarabine与TRAIL联合处理时对A549 细胞及人脐静脉内皮细胞(HUVEC,人正常细胞作为对照)增殖活力的影响,流式细胞术检测A549 细胞凋亡情况,Western blot法检测凋亡相关蛋白Survivin、Bcl-2、Bcl-xl、caspase-8、caspase-3、JNK和p38的表达情况。结果:25、50 μmol/LFludarabine或50 ng/mL TRAIL单独处理时对A549及HUVEC细胞增殖活力均无明显影响(P 均>0.05),两者联合处理后对A549细胞增殖活力抑制率分别为30.9%和44.9%。Fludarabine或TRAIL单独使用时对A549细胞凋亡无明显影响,联合处理时可使A549细胞凋亡率达到89.3%。与对照组、Fludarabine组和TRAIL组相比,Fludarabine与TRAIL联合处理可使A549细胞中Survivin、Bcl-2、Bcl-xl表达降低(P 均<0.05),促进caspase-8及caspase-3的激活,并能促进JNK和p38的磷酸化(P 均>0.05)。结论:Fludarabine与TRAIL联合处理可促进A549细胞凋亡。

OBJECTIVE: This study aimed to investigate whether Fludarabine, at a low-concentration, would enhance induction of apoptosis in A549 cells by TRAIL. METHODS: A549 cultures were organized into different groups: control, Fludarabine, TRAIL, and Fludarabine plus TRAIL. The CCK- 8 method was used to detect effect of Fludarabine and/or TRAIL on A549 and human umbilical vein endothelial cells (HUVEC). Flow cytometry was used to detect apoptosis rates. Western Blot was used to detect apoptosis-related proteins Survivin, Bcl-2, Bcl-xl, caspase-8, caspase-3, and the expression of JNK and p38. RESULTS: Fludarabine(25 or 50 μmol/L) or TRAIL alone had no significant effects on viability of A549 and HUVEC cells, or apoptosis in A540 cells. The inhibition rates from combined treatment of Fludarabine (25 or 50 μmol/L) and TRAIL were 30.9% and 44.9%, respectively, in A549 cells and the effect was concentration dependent. The combined treatment also caused apoptosis in 89.3% of A549 cells, as well as decreased expression of Survivin,Bcl-2 and Bcl-xl,promoted the activation of caspase-8 and caspase-3,and promoted the phosphorylation of JNK and p38. CONCLUSION:Fludarabine combined with TRAIL specifically induced apoptosis in A549 cells.