目的 探讨三氧化二砷(ATO)联合阿克拉霉素(ACM)对急性髓系白血病细胞株KG-1a的协同细胞杀伤效应及其分子机制.方法 采用克隆形成实验检测不同浓度ATO和ACM对KG-1a细胞增殖的影响,采用Compusyn软件分析两药联合是否具有协同作用,采用瑞氏染色法和流式细胞术分析两药联合对KG-1a细胞诱导凋亡作用,采用Western blot法分析两药联合后凋亡相关蛋白表达的差异.结果 ATO和ACM单药对KG-1a细胞具有生长抑制作用,且呈剂量依赖性增加.流式细胞术检测结果显示,0.4 μmol/L ATO+10 nmol/L ACM处理48 h后,KG-1a细胞的凋亡率为(34.5±3.1)%,与0.4 μmol/L ATO单药组和10 nmol/L ACM单药组[分别为(7.6±1.1)%和(18.7±2.3)%]比较,差异均有统计学意义(均P<0.05).1.5 μmol/L ATO+37.5 nmol/L ACM组KG-1a细胞的凋亡率为(52.5±4.7)%,与1.5 μmol/L ATO单药组和37.5 nmol/L ACM单药组[分别为(19.1±3.2)%和(27.7±2.2)%] 比较,差异均有统计学意义(均P<0.05).3.0 μmol/L ATO+75 nmol/L ACM组KG-1a细胞的凋亡率为(61.3±4.5)%,与3.0 μmol/L ATO单药组和75 nmol/L ACM单药组[分别为(29.5±2.5)%和(28.6±3.4)%]比较,差异均有统计学意义(均P<0.05).与单药组比较,联合组KG-1a细胞的细胞质空泡化、细胞皱缩和细胞核收缩等细胞凋亡的形态学变化更明显.Compusyn软件分析结果显示,联合药物处理组的联合指数均<1,表明两药联合具有协同细胞杀伤作用.结论 ATO联合ACM对急性髓系白血病细胞株KG-1a具有协同细胞杀伤作用,该协同作用可能通过激活凋亡相关信号通路,抑制KG-1a细胞的增殖并诱导凋亡.%Objective To investigate the synergistic lethal effect and mechanism of arsenic trioxide (ATO) and aclacinomycin (ACM) on human acute myeloid leukemia cell line KG-1a.Methods Colony-forming assay was used to detect the proliferation of KG-1a cells treated with different concentration of ATO and ACM.Compusyn software was used to analyze the synergistic effect of ATO and ACM.Flow cytometry and Wright′s staining were used to analyze the apoptotic rate of KG-1a cells induced by combined treatment of ATO and ACM.Western blot was used to determine the expression of proteins associated with apoptosis.Results The cytotoxicity of arsenic trioxide or aclacinomycin alone was in a dose-dependent manner.Flow cytometry analysis showed that the apoptotic rate of KG-1a cells treated with both 0.4 μmol/L ATO and 10 nmol/L ACM was (34.5±3.1)%, significantly higher than (7.6±1.1)% of 0.4 μmol/L ATO treatment or (18.7±2.3) % of 10 nmol/L ACM treatment alone (P<0.05).The apoptotic rate of KG-1a cells treated with both 1.5 μmol/L ATO and 37.5 nmol/L ACM was (52.5±4.7)%, significantly higher than (19.1±3.2)% of 1.5 μmol/L ATO treatment or (27.7±2.2)% of 37.5 nmol/L ACM treatment alone (P<0.05).The apoptotic rate of KG-1a cells treated with both 3.0 μmol/L ATO and 75 nmol/L ACM was (61.3±4.5)%, significantly higher than (29.5±2.5)% of 3.0 μmol/L ATO treatment or (28.6±3.4) % of 75 nmol/L ACM treatment alone (P<0.05).In addition, the result of Wright′s staining showed that combined treatment of ATO and ACM induced a more apparent phenotype of apoptosis when compared with single agent treatment.Compusyn software analysis showed that the combination index (CI) value of combined treatment group was less than 1, which indicated the synergistic effect of these two agents.Conclusions Combined treatment of ATO and ACM shows a synergistic lethal effect on human acute myeloid leukemia cell line KG-1a via activating the apoptotic pathway, which inhibits cell growth and induces apoptosis.
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