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

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Beclinl、凋亡抑制基因SurvivinmR．NA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（Beclinl、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24h、48h、72h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（尸〈0．05），Bedin1、SurvivinmRNA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BeclinlmRNA表达，而使Survivin表达受抑制有关．

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

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.