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

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

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。

Aim To detect the anti-tumor effects and mechanisms of 0-（ 4-ethoxiyl-butyl ）-berbamine （EBB） , a derivative of bisbenzylisoqunoline alkaloid, on reactive oxygen species （ROS）, apoptosis and cytosolic Ca2＋ （ [ Ca2 ± ]i ） in muhidrug 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 fluores- cence spectrophotometer. The [ Ca2＋]i was observed with laser scanning confocal microscopy. Results EBB showed obvious growth inhibiting effect on K562/ A02（8.22 ±1.67）μmol · L^ -1andK562（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 of [ Ca2＋ ]i in both cell lines. The [ Ca2＋ ]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 and [ Ca2 ＋ ]i are important for the effects of EBB.