阿糖胞苷诱导HL-60细胞凋亡的分子机制探讨  

Study on mechanism of apoptosis induced by Ara-C in leukemia HL-60 cells

作　　者：王甦[1] 刘云鹏[1] 候科佐[1] 罗颖[1] 刘世洲[1] 王妍[1] 

机构地区：[1]中国医科大学第一附属医院肿瘤内科

出　　处：《临床肿瘤学杂志》2008年第3期197-200,共4页Chinese Clinical Oncology

基　　金：辽宁省科技攻关项目(2004225004-11)

摘　　要：目的：探讨阿糖胞苷（Ara-C）诱导HL-60细胞凋亡的分子机制。方法：采用苔盼蓝拒染法检测细胞活力，MTF法测定细胞生长抑制率；形态学观察和流式细胞仪检测细胞凋亡；Western blot方法检测Bcl-2和caspase-3及其裂解片段的蛋白表达。结果：1～100斗M Ara-C以时间和剂量依赖方式抑制HL-60细胞增殖，诱导细胞凋亡，并下调Bcl-2蛋白表达，激活caspase-3。同时加入MEK抑制剂PD98059明显增加Ara-C的细胞毒作用和对caspase-3的激活作用，且这种作用可被P38MAPK抑制剂SB203580拮抗；PD98059和SB203580二者均未影响Ara-C对Bcl-2蛋白的下调作用。JNK抑制剂SP600125对Ara-C的上述作用无明显影响。结论：Ara-C以时间和剂量依赖方式抑制HL—60细胞增殖，并诱导细胞凋亡，同时下调Bcl-2蛋白表达，激活caspase-3；MEK抑制剂正性调节Ara-C的细胞毒作用，P38MAPK抑制剂负性调节Ara-c的细胞毒作用，二者可能是在caspase-3水平而未改变Bcl-2水平来发挥作用的。Objective：To explore the machanism of apoptosis induced by Ara-C in HL-60 cells. Methods：Cell viability was determined by trypan blue exclusion ,the growth inhibition was analyzed by MTY assay; cell apoptosis by morphology and flow cytometry; Bcl-2 and caspase-3 by western blot. Results：Ara-C （1-100μM） inhibited cellular proliferation and induced apoptosis and reducde the levels of Bcl-2 proteins and activated caspase-3 in a dose and time dependent fashion in HL-60 cells. PD98059 （25 μM） （ a inhibitor of MEK） enhanced apoptosis and activity of caspase-3 induced by Ara-C without altering Bcl-2 protein. However, SB203580 （20μM） （a inhibitor of P38MAPK） reduced the apoptosis and activity of caspase-3 induced by Ara-C without altering Bcl-2 protein. SP600125 （a inhibitor of JNK） did not affected effects of Ara-C. Conclusion：Ara-C downregulates Bcl-2 and activates caspase-3 during apoptosis in HL-60 cells; a inhibitor of MEK enhances apoptosis induced by Ara-C. However,a inhibitor of P38MAPK reduces the effects of Ara-C in HL-60 cells.

关 键 词：MAPK 抑制剂 阿糖胞苷 BCL-2 CASPASE-3 凋亡 

分 类 号：R730.53[医药卫生—肿瘤]