过氧化物酶体增殖物激活受体γ激动剂对白血病HL-60细胞的体外增殖抑制作用及机制的探讨  

Anti-proliferation effect of peroxisome proliferator activated receptor γ agonist on HL-60 cells in vitro and its mechanisms of action

作　　者：王春芝[1] 刘晓丹[1] Prem Raj-Shrestha 肖若芝[1] 林东军[1] 黄仁魏[1] 刘加军[1] 

机构地区：[1]中山大学附属第三医院血液科血液病研究所,广州510630

出　　处：《中华临床医师杂志（电子版）》2010年第9期94-98,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(30570786);教育部新世纪优秀人才支持计划资助项目(NCET-0721);广东省自然科学基金(8151008901000128)

摘　　要：目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)激动剂匹格列酮(PGZ)对白血病HL-60细胞的体外增殖抑制作用及其作用机制。方法以不同浓度的PGZ(20～80μmol/L)作用于体外培养的HL-60细胞24h、48h及72h,MTT法检测细胞生长抑制率,流式细胞术(FCM)检测细胞周期及细胞凋亡。应用RT-PCR及免疫印迹法(Westernblot)检测药物作用后PPARγ、Caspase-3及其裂解底物多聚(ADP-核糖)聚合酶(PARP)表达水平的变化。结果 PGZ可显著抑制细胞的生长及诱导细胞发生凋亡,呈现出明显的量-效与时-效关系。FCM检测结果表明,细胞主要被阻滞在G0/G1期,60μmol/L的PGZ作用48h后可以出现典型的亚G1期峰(细胞凋亡峰),PGZ在诱导细胞凋亡的同时,PPARγmRNA及蛋白的表达水平均逐渐升高。RT-PCR表明,Caspase-3酶原及其作用底物PARP表达水平逐渐降低,Westernblot结果显示32kD的Caspase-3酶原被活化出现17kD亚单位片段,同时Caspase-3的底物PARP被裂解出现89kD的亚单位片段。结论 PGZ在体外对HL-60细胞具有显著的细胞周期阻滞作用,并诱导细胞发生调亡。通过依赖PPARγ信号途径以及激活Caspase-3可能是PGZ抑制细胞增殖及诱导细胞发生凋亡的重要作用机制之一。Objective To investigate the apoptosis inducing effect of peroxisome proliferator activated receptor γ （PPARγ） agonist pioglitazone （PGZ） on leukemic HL-60 cells in vitro and its mechanisms of action.Methods HL-60 cells in culture medium in vitro were given different concentrations of PPARγ agonist PGZ （20-80 μmol/L） for 24,48 and 72 h.The inhibitory rate of the cells were measured by MTT assay,cell cycle and apoptosis was detected by flow cytometry （FCM）.RT-PCR and Western blot were used to detect both mRNA and protein levels of PPARγ as well as caspase-3 and poly（ADP-ribose） polymerase （PARP）.Results PGZ could inhibit the growth and cause apoptosis in HL-60 cells remarkably,the suppression was both in time-and dose-dependent manner.FCM detection revealed that the cells were mainly arrested in G0 /G1 phase and the subG1 cells were also found clearly in the histogram.Both mRNA and protein levels of PPARγ were upregulated concomitantly when apoptosis occurred.RT-PCR revealed that both caspase-3 zymogen and PARP were downregulated,whereas Western blot showed cleavage of the caspase-3 zymogen protein （32 kD）,with the appearance of its 17 kD subunit,and a cleaved 89-kD fragment of 116 kD PARP after the cells were treated by PGZ.Conclusions PGZ demonstrates anti-leukemia effect in HL-60 cells in vitro by inducing cell cycle arrest and apoptosis.Dependent on PPARγ signaling pathway and activation of caspase-3 may be one of its most important mechamisms

关 键 词：白血病 过氧化物酶体增殖物激活受体 细胞凋亡 匹格列酮 

分 类 号：R733.7[医药卫生—肿瘤]