DNA引物酶抑制剂碘化-3,3′-二乙基-9-甲基-硫杂羰花青诱导人白血病HL-60细胞凋亡(英文)  

Apoptosis induced by DNA primase inhibitor 3,3′-diethyl-9-methylthia-carbocyanine iodide in human leukemia HL-60 cells

作　　者：李志铭[1] 姜文奇[1] 管忠震[1] 朱孝峰[1] 周军民[1] 谢冰芬[1] 冯公侃[1] 朱振宇[1] 刘宗潮[1] 

机构地区：[1]中山大学肿瘤防治中心华南肿瘤学国家重点实验室,广东广州510060

出　　处：《药学学报》2006年第10期978-984,共7页Acta Pharmaceutica Sinica

基　　金：National Natural Science Foundation of China(39870886,30371621 and 30400589).

摘　　要：目的研究DNA引物酶抑制剂碘化-3,3′-二乙基-9-甲基-硫杂羰花青(DMTCC I)诱导人粒细胞性白血病HL-60细胞凋亡并探索其机制。方法分别采用不同浓度的DMTCC I处理培养于RPM I-1640培养基的HL-60细胞。采用MTT法检测DMTCC I对HL-60细胞的生长抑制作用。采用流式细胞仪和DNA琼脂糖凝胶电泳方法检测细胞凋亡。采用蛋白免疫印迹(W estern b lotting)法观察凋亡相关蛋白survivin,Bc l-xL,Bad,Bax,Bc l-2,caspase-9,caspase-3,caspase-6,PARP,DFF45和lam in B的表达。采用ApoA lert Caspase-3分析试剂盒检测caspase-3的活性。结果DMTCC I具有抑制人白血病HL-60细胞增殖的作用,其IC50值为0.24μmol.L-1。流式细胞仪和DNA琼脂糖凝胶电泳结果显示,DMTCC I可诱导HL-60细胞凋亡。在经DMTCC I处理的HL-60细胞中,survivin和Bc l-xL蛋白的表达水平下调,Bad和Bax蛋白的表达水平上调,Bc l-2蛋白的表达水平无变化,caspase-9,caspase-3,caspase-6,PARP,DFF45和lamin B被分别裂解,产生相应裂解产物。在HL-60细胞中,caspase-3的活性在1μmol.L-1DMTCCI处理3 h时明显升高,在处理12 h时达到最高峰。结论DMTCCI可抑制人白血病HL-60细胞的增殖并诱导其发生细胞凋亡。Bcl-2家族蛋白、survivin和caspases家族蛋白可能参与了上述诱导HL-60细胞凋亡的过程。Aim To investigate apoptosis induced by 3,3＇-diethyl-9-methylthia-carbocyanine iodide （DMTCCI） , an inhibitor of DNA primase found in our previous study, and the mechanism of DMTCCI in human myelogenous leukemia HL-60 cells. Methods HL-60 cells were cultured in RPMI-1640 medium and treated with different concentrations of DMTCCI. MTT assay was used to detect growth inhibition. Flow cytometry and DNA ladders were used to detect apoptosis. Western blotting was used to observe the expression of survivin, Bcl-xL, Bad, Bax, Bcl-2, caspase-9, caspase-3, caspase-6, PARP, DFF45 and lamin B protein. Caspase-3 activity was measured by ApoAlert Caspase-3 Assay Kit. Results DMTCCI inhibited proliferation of human leukemia HL-60 ceils with IC50 value of 0.24 μmol· L ^-1 The results of flow cytometry and DNA ladders showed that DMTCCI could induce apoptosis of HL-60 cells. The expression levels of protein survivin and Bcl-xL were down-regulated, Bad and Bax were up-regulated, while Bcl-2 protein had no change in response to DMTCCI treatment in HL-60 cells. Treatment of HL-60 cells with DMTCCI induced the proteolytic cleavage of caspase-9, caspase-3, caspase-6, PARP, DFF45 and lamin B protein. Caspase-3 activity apparently increased at 3 h and reached a peak at 12 h after exposure to 1 μmol · L ^-1 of DMTCCI in HL-60 cells. Conclusion DMTCCI inhibited proliferation and induced apoptosis of human leukemia HL-60 cells. Bcl-2 family proteins, survivin and caspases family proteins might play a role in the apoptosis process induced by DMTCCI.

关 键 词：DNA引物酶 DMTCCI 细胞凋亡 HL-60 

分 类 号：R963[医药卫生—微生物与生化药学]