藤黄酸对白血病K562/A02细胞的耐药逆转作用  

Reversal Effect of Gambogic Acid on Multidrug Resistance of K562/A02 Cell Line

作　　者：田亮[1,2] 刘娟[2] 陈宝安[1,2] 程坚[1] 丁家华[1] 王帅[1] 夏国华[1] 高峰[1] 邵泽叶[1] 张海军[1,2] 郭青龙[3] 张海伟[2,3] 王磊[1,2] 任艳艳[1,2] 蔡晓辉[1,2] 刘苒[1,2] 

机构地区：[1]东南大学医学院中大医院血液肿瘤科,江苏南京210009 [2]东南大学医学院临床肿瘤学系,江苏南京210009 [3]中国药科大学,江苏南京210009

出　　处：《中国实验血液学杂志》2012年第2期252-257,共6页Journal of Experimental Hematology

基　　金：国家重点基础研究发展计划(973计划)子课题(编号2010CB732404);国家自然科学基金(编号81170492);国家自然科学基金(编号30872970);江苏省重点学科资助

摘　　要：本研究旨在探讨藤黄酸(gambogic acid,GA)对K562/A02细胞株的耐药逆转作用及其逆转机制。采用MTT法检测GA对K562和K562/A02细胞的增殖抑制作用及对K562/A02细胞阿霉素(ADM)耐药逆转效应;用流式细胞术检测GA联合ADM对K562及K562/A02细胞凋亡率的影响;DAPI荧光染色观察ADM联合GA作用后细胞的形态学改变;Western blot法检测K562及K562/A02细胞P-糖蛋白(P-gp)、存活蛋白(Survivin)基因的表达。结果表明:ADM作用48 h抑制K562和K562/A02细胞增殖的IC50值分别为(1.42±0.07)μg/ml和(28.42±1.40)μg/ml。GA≤0.0625μmol/L时,对K562及K562/A02细胞株无明显增殖抑制作用;0.0625μmol/L GA联合ADM作用于K562/A02细胞能增加其对ADM的敏感性,耐药逆转倍数为1.53。0.0625μmol/L GA联合ADM作用于K562/A02细胞48 h能提高细胞的凋亡率(P<0.05),下调Survivin及P-gp蛋白的表达(P<0.05)。结论:GA可以逆转K562/A02细胞的耐药性,增强耐药细胞对ADM的敏感性,其机制可能与提高K562/A02细胞凋亡、下调Survivin和P-gp蛋白的表达有关。This study was purposed to investigate the reversal effect of gambogic acid（GA） on multidrug resistance of K562/A02 cells and its mechanism.The IC50（half maximal inhibitory concentration） of adriamycin（ADM） was evaluated by MTT.Cell apoptosis was detected by flow cytometry.Morphological changes of K562/A02 cells were observed by fluorescent microscopy with DAPI staining.The expressions of Survivin and P-gp were determined by Western blot.The results showed that the IC50 of ADM on K562 and K562/A02 cell proliferation were（1.42±0.07） μg/ml and（28.42±1.40） μg/ml respectively.GA≤0.0625 μmol/L had no inhibitory effect on proliferation of K562 and K562/A02.0.0625 μmol/L GA could enhance the sensitivity of K562/A02 cells to ADM（P0.05） and the reversal multiples was 1.53.The apoptotic rate was raised after treating with ADM combined with 0.0625 μmol/L GA for 48 h（P0.05）.Morphological differences were typical and obvious between cells of control and treated groups under fluorescence microscopy using DAPI staining.After treating K562/A02 cells with ADM combined with 0.0625 μmol/L GA for 48 h,the expressions of Survivin and P-gp were down-regulated at protein levels.It is concluded that GA can enhance the sensitivity of K562/A02 cells to ADM,which may be related to increasing cell apoptosis and down-regulating expressions of Survivin and P-gp.

关 键 词：藤黄酸 阿霉素 K562细胞 K562/A02细胞 多药耐药 

分 类 号：R733.7[医药卫生—肿瘤]