塞来昔布逆转MCF-7/Adr细胞多药耐药及其机制探讨  

Multidrug resistance reversal effect of celecoxib in human breast cancer cell line MCF-7/Adr and its mechanism

作　　者：陈琛[1] 许文林[1] 秦茹娟[1] 房新建[1] 方莉莉[1] 陈巧云[1] 

机构地区：[1]江苏大学附属人民医院中心实验室,江苏镇江212002

出　　处：《实用肿瘤杂志》2009年第3期237-242,共6页Journal of Practical Oncology

基　　金：江苏省卫生重大课题基金资助项目(K2005017)

摘　　要：目的观察环氧合酶-2(COX-2)抑制剂塞来昔布对肿瘤多药耐药(MDR)的逆转作用,并探讨其初步作用机制。方法在乳腺癌耐药细胞株MCF-7/Adr中,应用MTT方法检测塞来昔布对多柔比星(阿霉素,DOX)的耐药逆转倍数;RT-PCR及Western blot方法检测基因及蛋白的变化;流式细胞术(FCM)检测细胞膜P-gp的表达、功能、细胞周期和凋亡。结果塞来昔布作用24小时后,MCF-7/Adr细胞对DOX的敏感性明显增加,浓度为10μmol/L及20μmol/L时,耐药逆转倍数分别为1.82和3.80,呈现剂量依赖性。COX-2、mdr1、c-Jun、YB-1、survivin等在基因和蛋白水平明显下调(P<0.01),NF-κB无明显差异(P>0.05);20μmol/L作用24小时,P-gp的功能受抑制;G0+G1期细胞增多,S期细胞减少(P<0.05),凋亡率明显增高(P<0.05)。结论选择性环氧合酶-2抑制剂塞来昔布可有效逆转MCF-7/Adr细胞多药耐药,在一定的浓度范围内呈现剂量依赖性,初步机制可能涉及干预转录因子的表达而下调mdr1,阻滞细胞周期及增加细胞凋亡。Objective To investigate the role of specific cyclooxygenase-2 （COX-2） inhibitor,celeeoxib,on multidrug resistance （MDR） reversal in human breast cancer cell line MCF-7/Adr,and to explore its mechanism. Methods MCF-7/ Adr cells were treated with celecoxib at different concentrations. MTT assay was used to assess the reversal effect of celecoxib on MCF-7/Adr cells. RT-PCR was performed on the celecoxib treated MCF-7/Adr cells to examine COX-2,mdrl,c-Jun,YB- 1, NF-kappaB, survivin mRNA expressions. Western blotting analysis was performed to detect the expression of proteins above mentioned. Flow cytometry （FCM） was applied to analyze the expression of P-glycoprotein （P-gp）. Rhodamine 123 efflux assay was used to detect P-gp function. Cell cycle kit and Annexin V kit were used to test the cell cycle and apoptosis of MCF-7/Adr cells treated with celecoxib. Results Celecoxib restored the chemotherapeutic effect of doxorubicin （DOX） on MCF-7/Adr in dose-dependent manner, when celecoxib （I0, 20 p.mol/L） were added into the culture medium, the chemosensitivity of DOX was increased to 1.82 and 3.80 folds （P〈0. 01）. Genes and proteins of COX-2, mdrl ,c-Jun, YB-1, survivin were down-regulated along with celecoxib treatment ,while NF-kappaB was not affected. Twenty-four hours after the treatment of celecoxib （20μmol/L）,the expression and function of P-gp were significantly reduced,the rate of the cells at G1 ＋G0 stage increased,while the rate of those at S stage was decreased markedly （P〈0.05） and apoptotic rate significantly increased （P〈（O. 01）. Conclusion Celecoxib has a MDR reversal effect on MCF-7/Adr cell,which is probably involved in enhancing the arrestment of cells at G1 ＋G0 phase,inhibiting mdrl expression as well as promoting cells apoptosis.

关 键 词：乳腺肿瘤 基因 MDR MCF-7/ADR细胞 塞来昔布 P糖蛋白 转录因子 

分 类 号：R96[医药卫生—药理学] R737.9[医药卫生—药学]