3-MA对阿霉素诱导K562、K562/ADM细胞凋亡及其相关基因mRNA表达的影响  

3-MA on Adriamycin Induced K562 and K562 / ADM Cells Apoptosis and Related Gene mRNA Expression

作　　者：王莉[1,2] 刘春艳[1,2] 张炜[2] 赵丽[2] 

机构地区：[1]兰州大学第一临床医学院,甘肃兰州730030 [2]兰州大学第一医院中心实验室,甘肃兰州730000

出　　处：《西北民族大学学报（自然科学版）》2013年第3期54-60,共7页Journal of Northwest Minzu University(Natural Science)

基　　金："中央高校基本科研业务专项基金"自由探索项目(lzujbky-2011-99)

摘　　要：目的:通过自噬抑制剂3-甲基腺嘌呤(3-MA)对阿霉素(ADM)诱导白血病K562细胞及K562/ADM细胞的细胞效应和自噬基因Beclin1、凋亡抑制基因Survivin mRNA表达的变化观察,探讨自噬在细胞凋亡中的作用和机制.方法:体外培养K562和K562/ADM细胞,采用MTT法分别检测ADM及3-MA预处理对K562、K562/ADM细胞增殖的影响,流式细胞仪检测细胞凋亡率,实时RT-PCR法检测细胞自噬及凋亡相关基因(Beclin1、Survivin)mRNA表达的变化.结果:ADM可抑制K562与K562/ADM细胞增殖,且抑制作用呈现浓度与时间依赖性.ADM诱导组K562与K562/ADM细胞在24 h、48 h、72 h细胞凋亡率均较空白对照组明显提高(P<0.05).在ADM诱导前,经3-MA预处理可使ADM诱导的K562与K562/ADM细胞抑制率和细胞凋亡率均较单用ADM显著提高(P<0.05),Beclin 1、Survivin mRNA相对表达量均较单用ADM明显下降(P<0.05),呈正相关(r=0.827,P<0.01).结论:ADM可抑制K562、K562/ADM细胞的生长,并诱导细胞凋亡.3-MA通过抑制细胞的自噬可增强ADM诱导白血病细胞K562、K562/ADM的凋亡,其机制可能与下调Beclin1 mRNA表达,而使Survivin表达受抑制有关.Objective To investigate autophagy in cell apoptosis and mechanisms through the autophagy inhibitor 3- methyladenine （3- MA） on the adriamycin （ADM） induced leukemia K562 cells and K562/ ADM cells effect and autophagy genes Beclinl, Survivin apoptosis inhibiting gene mRNA expression change of observation. Methods In vitro cultivation of K562 and K562/ADM cells, Determined by MTT method was used respectively to detect the ADM and 3 - MA pretreatment effect on K562 and K562/ADM cells proliferation, Cell apoptosis was detected by flow cytometry, Real - time RT- PCR assay autophagy and apoptosis-related genes （Beclinl, survivin） mRNA expression changes. Results ADM inhibited K562 and K562/ADM cells proliferation, and the inhibition concentration and time dependence. In ADM induction group, K562/ADM K562 cells apoptosis rate at 24 h, 48 h, 72 h were greater significantly than the control group （P ~ 0.05）. Before the ADM induction, 3 - MA pretreatment could make the ADM - induced K562 and K562/ADM cells inhibition rate and apoptosis rate were higher significantly than that induced with only ADM （P 〈 0.05）. The relative expression levels of BediM, survivin mRNA decreased significantly when compared to only ADM induction （P 〈 0.05）, with a positive correlation （ r = 0. 827, P ~ 0.01 ）. Conclusions ADM inhibited the growth of K562 and K562/ADM cells, and induced cell apoptosis. 3 - MA cells by inhibiting autophagy can enhance the ADM induced leukemia cell K562, K562/ADM apoptosis. The mechanism may be associate to lower Bedim mRNA expression, and such suppressed the expression of survivin.

关 键 词：3-甲基腺嘌呤 K562细胞 K562 ADM细胞凋亡 诱导因素 

分 类 号：R979.14[医药卫生—药品]