摘要
目的:观察Aurora激酶抑制剂ENMD-2076对人急性髓系白血病(AML)细胞株的杀伤作用。方法:将不同浓度的ENMD-2076分别作用于对数生长期的AML细胞24 h、48 h,采用MTT比色法检测药物对细胞的生长抑制作用;应用Hoechst荧光染色法检测细胞凋亡率,Western blot法检测药物处理后Caspase途径和凋亡相关蛋白的改变。结果:ENMD-2076能显著抑制AML细胞株THP-1和Kasumi-1的生长(P<0.001),24 h时对THP-1和Kasumi-1的半数抑制浓度分别是9.32μmol/L和7.23μmol/L。Hoechst荧光染色证实ENMD-2076能使THP-1细胞发生染色质浓缩等凋亡改变。Western blot检测证实,ENMD-2076能激活Caspase-3、Caspase-9、Caspase-8和PARP,上调促凋亡蛋白Bak、Bax、Bad和Bim,下调抗凋亡蛋白Mcl-1的表达。结论:Aurora激酶抑制剂ENMD-2076能显著抑制人AML细胞株THP-1和Kasumi-1的增殖并促进其凋亡,对凋亡相关蛋白的调控是其作用机制之一。
Objective： To investigate the effect of aurora kinase inhibitor ENMD-2076 on human acute myelogenous leukemia（AML） cell lines.Methods： AML THP-1 and Kasumi-1 cells were treated with ENMD-2076 for 24 h and 48 h,respectively.Cell growth was measured by MTT assay.Apoptosis was determined using Hoechst staining apoptosis detection kit.Activation of Caspase pathway and expression of apoptosis regulator proteins were detected by Western blot.Results： ENMD-2076 significantly induced growth arrest and apoptosis in THP-1 and Kasumi-1 cells.Enhanced apoptosis was observed in ENMD-2076 group evidenced by strong activation of Caspase-9,Caspase-3 and PARP.Furthermore,the ENMD-2076 treatment resulted in down-regulation of anti-apoptotic protein Mcl-1 expression.Also,up-regulated expression of pro-apoptotic protein Bak,Bad and Bax was detected after ENMD-2076 treatment.Conclusion： ENMD-2076 can kill effectively AML cells by inhibiting cell growth and inducing apoptosis,which is associated with activation of Caspase pathway and regulation of pro-apoptotic and anti-apoptotic proteins.
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2012年第5期479-484,共6页
Journal of Zhejiang University(Medical Sciences)
基金
浙江省自然科学基金杰出青年团队(No.R2090392)资助项目
浙江理工大学科研启动基金(No.1016834-Y)
