盐霉素抑制耐格列卫的人慢性粒细胞白血病细胞株K562/Glv增殖并诱导其凋亡  

Salinomycin inhibits proliferation and induces apoptosis of Gleevec-resistant chronic myeloid leukemic cell line K562/Glv

作　　者：徐霜清[1,2] 祝爱珍[3] 刘成成[3] 许婷婷[3] 陈小宇[3] 刘革修[3] 

机构地区：[1]中南大学湘雅医学院检验系,湖南长沙410013 [2]广州金域医学检验中心,广东广州510330 [3]暨南大学医学院血液病研究所,广东广州510632

出　　处：《中国病理生理杂志》2012年第7期1208-1212,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30670902)

摘　　要：目的:探讨盐霉素对耐格列卫的人慢性粒细胞白血病细胞株K562/Glv抑制增殖和诱导凋亡的作用及其机制。方法:采用CCK-8的方法检测盐霉素对K562/Glv细胞生长的抑制作用;流式细胞术检测细胞凋亡、活性氧、细胞内Ca2+浓度([Ca2+]i)和线粒体膜电位(ΔΨm);比色法检测caspase-3、-8和-9活性;Westernblotting分析细胞色素C、Bcl-2、Bax、β-catenin和磷酸化低密度脂蛋白受体相关蛋白6(p-LRP6)蛋白水平。结果:盐霉素对K562/Glv细胞生长具有剂量依赖性抑制作用,0.2μmol/L时细胞增殖抑制率为(36.70±2.31)%,细胞凋亡率为(19.66±2.43)%;0.2μmol/L盐霉素作用于K562/Glv细胞,ΔΨm显著下降,24 h时下降至对照组的(19.8±2.4)%,细胞内活性氧和[Ca2+]i在短期显著升高。Caspase-3、-8和-9活性均显著增加,与对照组比较,差异有统计学意义(P<0.01)。Bcl-2的表达下调,Bax的表达明显增加,Bcl-2/Bax比值显著降低。同时,盐霉素也减少K562/Glv细胞内β-catenin和p-LRP6蛋白水平。结论:盐霉素不仅通过Bcl-2/Bax途径和线粒体凋亡途径诱导耐格列卫人慢性粒细胞白血病细胞K562/Glv的凋亡,而且通过抑制Wnt信号途径抑制K562/Glv细胞增殖。AIM： To study the effect of salinomycin on inhibiting proliferation and inducing apoptosis of Gleevec -resistant chronic myeloid leukemia cell line K562/GIv. MIgTRODS： The inhibitory effect of salinomycin on the growth of K562/Glv ceils was detected by CCK-8 assay in vitro. Flow eytometry was used to observe apoptosis, mitochon- dria membrane potential （△ψm）, reactive oxygen species （ROS） and the concentration of intracellular Ca2 ＋ （ [ Ca2 ＋ ]i） in K562/GIv cells. The activity of easpase-3, -8 and -9 was measured by the method of colorimetry. The levels of cyto- chrome C, Bel - 2, Bax, β - catenin and phosphorylated low - density lipoprotein receptor - related protein 6 （ p - LRP6） were determined by Western blotting. RESULTS： Salinomycin inhibited the growth of KS62/GIv cells in a dose -dependent manner. Sallnomycin at concentration of 0.2 μmol/L inhibited the growth of the cells with the inhibitory rate of （36.70 ± 2. 31 ） %. The cell apoptotic rate was （ 19.66 ± 2.23） %. Salinomycin at concentration of 0. 2 μmol/L decreased the level of △ψm, and increased the levels of ROS, cytochrome C and [ Ca^2 ＋ ] in the cells. Salinomycin also increased the activity of caspase - 3, - 8 and - 9 in the cells, reduced the ratio of BcI - 2/Bax, and attenuated the levels of 13 - catenin and p - LRP6. CONCLUSION： Salinomycin induces the apoptosis of Gleevec - resistant myeloid leukemia cell line K562/GIv via Bcl- 2/Bax and mitochondria-dependent pathways, and inhibits the cell growth through Wnt signal pathway.

关 键 词：盐霉素 格列卫 白血病 细胞凋亡 线粒体 β—catenin 低密度脂蛋白受体相关蛋白6 

分 类 号：R743[医药卫生—神经病学与精神病学]