摘要
目的探讨左卡尼汀作为TRAIL(tumor necrosis factor-related apoptosis inducing ligand)敏化剂,增强TRAIL对神经胶质瘤细胞诱导凋亡的效果,并对其敏化作用的机制进行研究。方法以U87为神经胶质瘤细胞模型,通过CCK-8检测细胞活性,Annexin V-FITC/PI染色、caspase-3表达及活性等指标检测细胞凋亡,通过RT-PCR、Western blot对NF-κB(nuclear factor kappa B)和c-FLIP(FLICE抑制蛋白)的转录与表达进行分析,沉默NF-κB,分析其与c-FLIP的关系。结果 TRAIL与左卡尼汀联用,癌细胞存活率明显下降,凋亡明显上升;联用组与对照组相比,c-FLIP的转录和蛋白表达以及NF-κB的转录均有明显降低;沉默NF-κB证明其为c-FLIP上游因子。结论左卡尼汀与TRAIL可以产生协同作用,诱导U87细胞凋亡,其敏化机制与抑制NF-κB信号通路以及其下游c-FLIP表达有关。
Aim To investigate the enhancing effect of L-carnitine as a sensitizer on tumor necrosis factor-re-lated apoptosis inducing ligand ( TRAIL)-induced ap-optosis in glioma cells. Methods Glioma cell U87 was used as model cell line. Cell viability was determined by CCK-8 , and apoptosis was assessed by Annexin V-FITC/PI staining, caspase-3 activity and expres-sion. The expression and transcription of nuclear factor kappa B ( NF-κB ) and FLICE inhibiting protein ( c-FLIP) were measured by RT-PCR and Western blot. In addition, NF-κB was knockdown to analyze its regu-lating effect on c-FLIP expression. Results The com-bination treatment with TRAIL and L-carnitine signifi-cantly inhibited cell proliferation and induced apopto-sis. Compared with control, combinational treatment significantly suppressed the transcription and expres-sion of c-FLIP as well as translocation of NF-κB. Through silencing NF-κB, NF-κB was found to act as upstream signaling to regulate c-FLIP. Conclusion L-carnitine sensitizes TRAIL-induced tumor cell apoptosis via suppression of NF-κB-dependent c-FLIP expres-sion.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第5期664-670,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81473384)
