钙调素拮抗剂O-(4-乙氧基-丁基)-小檗胺对人白血病细胞K562和K562/A02生长的抑制作用

目的研究钙调素拮抗剂O-(4-乙氧基-丁基)-小檗胺(EBB)体外抗白血病作用及机制。方法四唑盐(MTT)比色法评价EBB的细胞毒作用;流式细胞术检测细胞凋亡;荧光分光光度计检测细胞ROS水平;激光共聚焦显微镜检测Ca2+]i含量。结果EBB明显抑制敏感K562和耐药K562/A02细胞增殖,IC50分别为(8.22±1.67)mol.L-1和(8.97±1.48)mol.L-1。EBB作用早期诱导凋亡,晚期引起坏死。K562/A02细胞的ROS静息值是K562细胞的2倍,EBB诱导两种细胞ROS生成呈浓度和时间依赖性,加入SOD明显降低坏死率。K562/A02细胞的Ca2+]i低于K562细胞。EBB能明显增加两种细胞的Ca2+]i。结论EBB明显抑制K562和K562/A02细胞增殖。信号分子Ca2+、ROS参与了EBB的细胞毒作用。

The Growth inhibiting effect of calmodulin antagonist O-(4-ethoxiyl-butyl)-berbamine on leukemia cells K562 and K562/A02

Aim To detect the anti-tumor effects and mechanisms of O-(4-ethoxiyl-butyl)-berbamine(EBB),a derivative of bisbenzylisoqunoline alkaloid,on reactive oxygen species(ROS),apoptosis and cytosolic Ca2+(Ca2+]i)in multidrug resistant(MDR)human leukemia cell K562/A02 and its parental cell K562 in vitro.Methods The cytotoxicity of EBB on K562 and K562/A02 cells was evaluated by MTT assay.Flow cytometry analysis was used to detect apoptosis and necrosis.The ROS was determined by fluorescence spectrophotometer.TheCa2+]i was observed with laser scanning confocal microscopy.Results EBB showed obvious growth inhibiting effect on K562/A02(8.22±1.67)μmol·L-1 and K562(8.97±1.48)μmol·L-1 cells.In both cell lines,EBB induced apoptosis at the early stage and led to necrosis at the late stage.ROS in K562/A02 cells was twice of that in K562 cells.EBB induced ROS production in both cell lines in a dose and time dependent manner.SOD significantly reduced the necrotic ratios.EBB induced an increase ofCa2+]i in both cell lines.TheCa2+]i in K562/A02 cells was lower than that in K562 cells.Conclusions EBB has obvious growth inhibiting effects on K562 and K562/A02 cells.ROS andCa2+]i are important for the effects of EBB.