中文摘要：

目的：研究抗真菌药物克霉唑作为钙激活性钾通道阻断剂对人类急性早幼粒细胞白血病（APL）细胞及骨髓基质细胞增殖和凋亡的影响。方法：通过台盼兰拒染计数法检测了钙激活性钾通道阻断剂克霉唑对人APL细胞株NB4、APL患者原代细胞和骨髓基质细胞生存活力的影响,流式细胞仪检测细胞周期变化及亚二倍体的出现,AnnexinV/PI试剂盒和DAPI荧光染色检测细胞的凋亡情况,激光扫描共聚焦显微镜实时监测[Ca2＋]i的变化。结果：克霉唑以浓度依赖方式显著抑制NB4细胞和APL原代细胞生长,在同样条件下骨髓基质细胞没有显著毒性。5μmol/L克霉唑作用72h后,细胞周期明显被阻滞于G0/G1期,S期显著减少,并出现亚二倍体峰。Annexin V/PI双标记和DAPI荧光染色证实克霉唑诱导NB4细胞和APL原代细胞凋亡,凋亡率分别为（30.5±3.9）%、（33.7±4.5）%。5μmol/L克霉唑引起[Ca2＋]i荧光比值显著增高。结论：钙激活性钾通道阻断剂克霉唑诱导APL细胞凋亡,这为治疗APL提供了新的有希望的治疗方法,其机制可能与克霉唑调节细胞内钙水平有关。

英文摘要：

Objective：To investigate the effect of the antifungal antibiotic clotrimazole,a inhibitor of the intermediate-conductance Ca2＋ -activated K＋ （IK） channel,on the cell cycle and apoptosis of human acute myeloid leukemia cells and bone marrow stormal cells. Method：Fresh bone marrow cells and bone marrow stormal cells from APL patients and APL cell lines NB4 were used for in vitro culture study. The effects of clotrimazole on cells survival were determined by Trypan Blue exclusion. Flow cytometry was used to examine cell cycle distribution. The apoptosis rate was analyzed by using Annexin V-FITC kit and DAPI assay. The level of intracellular free Ca2＋ were observed with confocal laser scanning microscope.Result：Clotrimazole significantly suppreses growth of NB4 and APL primary cells in a concentration dependent manner. In contrast,there was no significant cytotoxicity at the same concentration to bone marrow stormal cells. DNA histograms showed a significantly larger percentage of cells in G0/G1 phase,a smaller one in S phase and the hypodiploid peak with 5 μmol/L clotrimazole treatment for 72 h. Clotrimazole induced apoptosis in NB4 and APL primary cells.The ratio of apoptosis cells was （30.5±3.9）% and （33.7±4.5）%,respectively. Clotrimazole increased the levels of [Ca2＋ ]i in NB4 and APL primary cells.Conclusion：Clotrimazole,a inhibitor of intermediate-conductance Ca2＋ -activated K＋ （IK） channel,induced apoptosis in APL,which provides a novel promising approach in the treatment of APL. The mechanism of growth-suppressive effect of clotrimazole was correlated with its ability to affect the levels of [Ca2＋]i.