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

作　　者：邹凌琳[1] 周圆[1] 杨铭[2] 代晓华[2] 齐静[2] 熊冬生[2] 范冬梅[2] 杨纯正[2] 朱惠芳[2] 

机构地区：[1]中国医学科学院北京协和医学院血液学研究所 [2]中国医学科学院北京协和医学院血液学研究所血液病医院实验血液学国家重点实验室,天津300020

出　　处：《中国药理学通报》2009年第10期1313-1317,共5页Chinese Pharmacological Bulletin

基　　金：高等学校博士学科点专项科研基金资助项目(No20070023093);天津市应用基础研究重点资助项目(No07JCZDJC04900)

摘　　要：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.

关 键 词：钙调素拮抗剂 O-(4-乙氧基-丁基)-小檗胺 人白血病细胞 活性氧 钙离子 凋亡 坏死 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]