摘要
目的:本研究旨在探讨survivin抑制剂YM155对人慢性髓系白血病细胞株K562凋亡和自噬的影响。方法:采用不同浓度YM155处理K562细胞,用CCK-8法检测细胞增殖抑制作用,流式细胞术检测细胞凋亡率,RTPCR检测survivin、BCL-2及beclin1的mRNA表达水平,Western blot检测survivin、BCL-2、caspase-3、PARP及LC-3的蛋白表达。结果:YM155抑制K562细胞的增殖,且呈时间及剂量依赖性。随着药物浓度的升高及作用时间的延长,survivin和BCL-2的mRNA及蛋白表达水平下降,而caspase-3、PARP、beclin1及LC-3的表达水平升高。YM155联合3-MA组的LC-3和caspase-3蛋白表达水平以及K562细胞凋亡率均显著低于YM155单用组。结论:YM155通过诱导凋亡和自噬而抑制K562细胞的增殖,其自噬诱导效应与凋亡诱导效应有协同抗CML的作用。
Objective： This study was purposed to investigate the effect of YM155, a survivin inhibitor, on the apop tosis and autophagy of K562 cells. Methods： K562 cells were treated with YM155 at different concentration. Cell sur- vival was analyzed by CCK-8 assay, the cell apoptosis was detected by flow cytometry. Survivin, BCL-2 and beclinl mRNA expressions were determined by RT-PCR. Survivin, BCL-2, caspase-3, PARP and LC-3 protein expressions were assayed by Western blot. Results： YM155 inhibited the proliferation of K562 cells in a time- and dose-dependent manners. With the increasing of YM155 concentration and prolonging of action time, the expression levels of mRNA and protein of survivin and BCL-2 decreased, while the expression levels of caspase-3, PARP, beclinl and LC-3 increased. Compared with the YM155 group, the protein levels of LC-3 and caspase-3 were lower in YM155 combined with 3-MA group. Conclusion： YM155 can inhibit K562 cell proliferation by inducing apoptosis and autophagy, while autophagy induction effect can enhance its cytotoxic effect.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2015年第2期375-380,共6页
Journal of Experimental Hematology
基金
江苏省淮安市科技支撑计划(HAS2013027)
