环氧合酶-2抑制剂联合苦参碱逆转K562/AO2细胞多药耐药的研究  

Effect of COX-2 inhibitor celecoxib,matrine and their combination on multidrug resistance of K562/AO2 cell line

作　　者：桂琳[1,2] 李静[1] 陈宝安[2] 高峰[2] 王筠[2] 蔡晓辉[2] 

机构地区：[1]东南大学医学院附属蚌埠第三人民医院血液科,安徽蚌埠233000 [2]东南大学附属中大医院血液科,江苏南京210009

出　　处：《东南大学学报（医学版）》2012年第2期174-178,共5页Journal of Southeast University(Medical Science Edition)

摘　　要：目的:研究塞来昔布单独及联合应用苦参碱对K562/AO2细胞多药耐药逆转作用的影响,以及探讨他们互相作用的机理。方法:采用四甲基偶氮唑盐(MTT)法检测阿霉素(ADM)的半数抑制量;流式细胞术检测细胞凋亡;RT-PCR方法检测多药耐药基因(MDR)和环氧合酶-2(COX-2)mRNA的表达水平;Westernblotting方法检测人p-糖蛋白(P-gp)和COX-2蛋白的表达水平。结果:ADM对K562、K562/AO2细胞的IC50值分别为0.398、33.31μg.ml-1;苦参碱(200μg.ml-1)、塞来昔布(7.5μg.ml-1)单独及联合应用处理细胞时,ADM对K562/AO2细胞的IC50值分别为9.44、12.84、2.71μg.ml-1;苦参碱、塞来昔布单独及联合应用于K562/AO2细胞后凋亡率明显增加,MDR1和COX-2 mRNA以及P-gp、COX-2蛋白的表达明显下调,且两药联合时下调更明显。结论:苦参碱和塞来昔布均有逆转K562/AO2细胞多药耐药的作用,两药联合应用时效果更加明显,P-gp的高表达可能与COX-2的表达上调有关。Objective：To investigate the effect of celecoxib alone and in combination with matrine on multidrug resistance of K562/AO2 cell line,and to explore the mechanism of their interaction.Methods：We use the methyl blue tetrazolium（MTT） assay to detect half the amount of adriamycin（ADM）.We have the detection of apoptosis by flow cytometry.We use RT-PCR to detect multidrug resistance（MDR） gene MDR1 and cyclooxygenase-2（COX-2） mRNA expression levels.We detected permeability glycoprotein（P-gp） and COX-2 protein expression by Western blotting.Results：ADM for K562,K562/AO2 the IC50 values were 0.398,33.31 μg·ml-1.When matrine（200 μg·ml-1）,celecoxib（7.5 μg·ml-1） alone and in combination-treated cells,the IC50 values of ADM for K562/AO2 were 9.44,12.84,2.71 μg·ml-1.Matrine,celecoxib alone or jointly used,K562/AO2 cell apoptosis was significantly increased,MDR1 and COX-2 mRNA and P-gp,COX-2 protein expression was significantly reduced.And the two-drug combination was significantly reduced.Conclusion：Matrine and celecoxib can reverse resistant K562/AO2 role,and when two drugs in combination was more evident.And high expression of P-gp may be associated with upregulation of COX-2.

关 键 词：塞来昔布 苦参碱 多药耐药 阿霉素 

分 类 号：R-331[医药卫生] R969.2