间变性淋巴瘤激酶抑制剂对人神经母细胞瘤细胞凋亡的影响

Effects on apoptosis of human neuroblastomas cells with anaplastic lymphoma kinase inhibitor TAE684

目的 观察间变性淋巴瘤激酶（ALK）抑制剂TAE684对人神经母细胞瘤SK-N-BE（2）细胞凋亡水平及凋亡蛋白表达的影响.方法 应用细胞计数试剂盒（CCK-8）检测不同TAE684浓度滴度（1、5、25、125、625 nmol/L）对体外培养的SK-N-BE（2）细胞作用的合适终质量浓度,Western blot检测TAE684对ALK蛋白和凋亡相关蛋白B细胞淋巴瘤/白血病-2（bcl-2）、B细胞淋巴瘤/白血病-2相关X蛋白（bax）和cleaved半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3表达的影响,流式细胞术检测细胞凋亡水平.结果 TAE684对SK-N-BE（2）细胞的半数抑制浓度（IC50）值为115 nmol/L,TAE684组ALK和bcl-2蛋白表达下调,分别为0.036±0.003和0.190±0.015;而bax和cleaved Caspase-3蛋白表达明显上调,分别为0.161 ±0.003和0.949±0.003,凋亡水平明显升高,为22.593 ±0.075.TAE684组与对照组和二甲基亚砜（DMSO）组比较,差异有统计学意义（P＜0.05）.结论 ALK抑制剂TAE684可能通过对凋亡分子的调控来实现对体外培养的SK-N-BE（2）细胞凋亡促进作用.

Objective To investigate the effects on the apoptosis and the expressions of some apoptosis-related proteins of human neuroblastomas cells with anaplastic lymphoma kinase （ALK） inhibitor TAE684.Methods Cell counting kit-8 （CCK-8） was used to detect different concentration of TAE684 （1,5,25,125,625 nmol/L） treated SK-N-BE （2） cells to analyze the appropriated concentration.The expressions of ALK protein,as well as B cell lymphoma/leukemia-2 （bcl-2）,B cell lymphoma/leukemia2 associated X protein （bax）,and cleaved Caspase-3 protein were detected by Western blotting,meanwhile,the apoptosis of human neuroblastomas （NB） cells were detected using flow cytometry.Results In vitro,the 50% inhibitory dose （IC50） value of TAE684 treated SK-N-BE （2） cells was 115nmol/L.The expressions of ALK and bcl-2 protein were down-regulated （0.036 ±0.003 and 0.190 ±0.015）.The expressions of bax and Cleaved Caspase-3 protein were up-regulated （0.161 ± 0.003 and 0.949 ± 0.003）.The flow cytometry showed that the cellular apoptosis levels were 22.593 ± 0.075.The differences among TAE684,the normal control group and DMSO group were significant （P 〈 0.05）.Conclusion ALK inhibitor TAE684 could induce the apoptosis of SK-N-BE （2） cells by regulating the apoptotic molecules.