Survivin反义寡核苷酸诱导K562细胞凋亡及化疗协同作用研究  

Survivin ASODN induced apoptosis of human leukemia cell K562 and synergistic effect with SN-38 in vitro

作　　者：封蔚莹[1,2] 周炀[1,2] 周国忠[1,2] 

机构地区：[1]绍兴市人民医院 [2]浙江大学绍兴医院血液科,浙江绍兴3120000

出　　处：《临床血液学杂志》2012年第4期454-456,共3页Journal of Clinical Hematology

摘　　要：目的:研究survivin反义寡核苷酸(ASODN)诱导慢性髓系白血病(CML)急变细胞系K562细胞凋亡及机制,并探讨与喜树碱(CPT)的活性代谢产物SN-38联合治疗的疗效。方法:Survivin ASODN转染K562细胞后,应用CCK-8法检测细胞增殖抑制率,流式细胞仪检测细胞凋亡情况,QRT-PCR法及Western blot法检测凋亡相关基因及蛋白的变化。依据Chou-Talalay中效原理,采用calcusyn软件计算两药联合指数(CI),评价survivin ASODN与SN-38联合治疗疗效,并以Western blot检测联合治疗后凋亡相关蛋白变化。结果:Survivin ASODN呈时间和剂量依赖性抑制K562细胞增殖,24、48、72h半数抑制率(IC50)分别约为600、498、296nmol/L。600nmol/L survivinASODN以时间依赖方式诱导K562细胞凋亡,下调survivin mRNA及蛋白表达,而Bcl-2蛋白表达无明显变化。survivin ASODN与SN-38联合治疗K562细胞48h,两药极低浓度时为拮抗及相加作用(Fa≤0.12),随浓度增大表现剂量依赖性的协同作用(Fa>0.12),两药最强协同效应时浓度为380nmol/L survivin ASODN和38nmol/L SN-38。联合组的survivin及Bcl-2蛋白表达较2单药组及对照组均明显下降。结论:survivin ASODN抑制K562细胞生长并诱导凋亡,和SN-38联合治疗通过下调survivin及Bcl-2表达发挥协同效应(Fa>0.12)。Objective：To observe the role of survivin ASODN in apoptosis of human chronic myeloid leukemia（CML）in blast crisis cell line K562 in vitro,and the effect of combination therapy with survivin ASODN and SN-38.Method：Inhibitive effect of survivin ASODN on cell growth was determined by CCK-8 test.Apoptosis was detected by Annexin V/PI kit through flow cytometry.Expression of apoptosis relative gene and protein was examined by QRT-PCR and western-blot.The combination index（CI）with survivin ASODN and SN-38 combination therapy was calculated by the method of Chou and Talalay using CalcuSyn software.Expression of apoptosis protein after combination therapy was detected by western blot.Result：Survivin ASODN inhibited cell proliferation of K562 in a dose-and time-dependent manner.The IC50 of K562 inhibited with survivin ASODN for 24 h,48 h or 72 h were 600,498 or 296 nmol/L,respectively.600 nmol/L survivin ASODN induced apoptosis of K562 cells in a time-dependent manner,with decrease of survivin mRNA and protein expression.But the expression of Bcl-2 did not change.The combination effect was additive and antagonism at lower concentration of survivin ASODN and SN-38（Fa≤0.12）,but was dose-dependent synergistic with higher concentration of two drugs（Fa〉0.12）.Combination therapy with 380 nmol/L survivin ASODN and 38 nmol/L SN-38 received the strongest synergistic antiproliferative effect.Survivin and Bcl-2 protein significantly decrease in combination group than survivin ASODN group or SN-38 group.Conclusion：Survivin ASODN can inhibit growth and induce apoptosis in human K562 cells.Survivin ASODN and SN-38 have synergistic effect on K562 cell by downregulating the survivin and Bcl-2（Fa〉0.12）.

关 键 词：白血病 SURVIVIN 反义寡核苷酸 

分 类 号：R733.7[医药卫生—肿瘤]