雷帕霉素协同去甲氧柔红霉素诱导人急性T淋巴细胞白血病Jurkat细胞株凋亡的作用  

Synergistic cytotoxic effects of rapamycin and idarubicin on human acute T-cell lymphoblastic leukemia Jurkat cells

作　　者：赵妍敏[1] 吴康妮[1] 王颖佳[1] 吴功强[1] 曹伟杰[1] 于晓红[1] 黄河[1] 

机构地区：[1]浙江大学医学院附属第一医院骨髓移植中心,浙江杭州310003

出　　处：《浙江大学学报（医学版）》2011年第5期482-488,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：研究者发起研究基金项目(CHN2009ONC007);国家自然科学基金资助项目(81000193)

摘　　要：目的:探讨去甲氧柔红霉素(idarubicin,IDA)联合雷帕霉素(rapamycin,Rapa)抗人急性T淋巴细胞白血病(T-ALL)Jurkat细胞株的效应及机制。方法:应用CCK-8法分析两药联用对细胞增殖的影响;Isobologram法分析两药联合作用的性质;电镜形态学观察和Annexin V/PI染色流式细胞仪检测细胞凋亡情况;免疫印迹法检测凋亡相关基因Caspase3、PARP、Caspase8、Caspase9及Akt、p-Akt、P85S6K、p-P85S6K、P70S6K、p-P70S6K、ERK1/2、p-ERK1/2等蛋白质水平表达。结果:CCK-8法示IDA联合Rapa能明显降低IDA的半数抑制浓度(50%inhibition concentration,IC50)值。Isobologram法示两药联合指数小于1。免疫印迹法示两药联用组较IDA单用组更能激活Caspase3和底物PARP,Caspase8和Caspase9在两药联用组均呈现明显激活。IDA联用Rapa后能使mTOR信号通路上游的Akt及下游的P85S6K、P70S6K分子磷酸化水平明显降低,并能协同下调ERK的磷酸化水平。结论:Rapa和IDA对Jurkat细胞的生长抑制具有协同作用,两药联用时细胞凋亡增加,且通过线粒体和细胞膜双途径增强凋亡效应。其协同机制可能与Rapa逆转IDA引起的Akt/mTOR信号通路激活,并抑制ERK信号活化有关。Objective: To investigate the cytotoxic effects of mTOR inhibitor rapamycin(Rapa) and idarubicin(IDA) on human T-cell acute lymphoblastic leukemia Jurkat cell line. Methods: The proliferation of Jurkat cells was observed by CCK-8 assay.The combined index was analyzed by Isobologram method.Apoptosis was detected by electron microscopy and flow cytometry with Annexin V/PI staining.Protein expressions of Caspase3,PARP,Caspase8,Caspase9,Akt,p-Akt,P85S6K,p-P85S6K,P70S6K,p-P70S6K,ERK1/2 and p-ERK1/2 were determined by Western blotting. Results: The IC50 of IDA for Jurkat cells was significantly reduced when combined with 10 nmol/L rapamycin.The combined index was <1.Both electron microscopy and Annexin V/PI staining flow cytometry revealed that rapamycin significantly increased apoptotic sensitivity to IDA.The combination of IDA with rapamycin enhanced the expressions of Caspase3,PARP,Caspase8 and Caspase9.Rapamycin significantly inhibited mTOR signaling upstream Akt and downstream S6K activation triggered by IDA,and the combination of the two agents led to synergistic inhibition of ERK phosphorylation. Conclusions: Combination of IDA with rapamycin exerted a synergistic anti-proliferative effect and promoted apoptosis by both extrinsic and intrinsic apoptotic pathways in Jurkat cells.Inhibition of ERK phosphorylation and mTOR signaling by rapamycin may play a certain role in this synergistic effect.

关 键 词：JURKAT细胞 白血病淋巴瘤 成人T细胞 西罗莫司/药理学 去甲氧柔红霉素/药理学 细胞凋亡/药物作用 信号传导 细胞增殖/药物作用 药物疗法 联合 

分 类 号：R733.71[医药卫生—肿瘤]