盐酸千金藤碱逆转K562/ADR细胞多药耐药性及其机制  

Correlation between reversing effect of cepharanthine hydrochloride on multidrug resistance and P-glycoprotein expression and function of K562/ADR cells

作　　者：彭有梅[1,2] 王宁[1,3] 王亚峰[2] 韩立[1,2] 张艳[1,3] 江金花[1,3] 周玉冰[1,2] 王庆端[1,3] 

机构地区：[1]郑州大学医药科学研究院 [2]郑州大学药学院 [3]河南省肝病药理重点实验室,河南郑州450052

出　　处：《药学学报》2012年第5期594-599,共6页Acta Pharmaceutica Sinica

基　　金：河南省基础与前沿技术研究计划项目(082300453204)

摘　　要：研究盐酸千金藤碱(cepharanthine hydrochloride,CH)逆转K562/ADR细胞多药耐药性及其机制。采用MTT法检测多柔比星(adriamycin,ADR)单用及分别与CH、维拉帕米(verapamil,VER)合用的细胞毒作用;采用流式细胞仪,测定CH对细胞内ADR蓄积、罗丹明123(Rho123)蓄积和泵出及P糖蛋白(P-gp)表达的影响。结果表明,CH(4μmol.L1)使K562/ADR细胞对ADR的敏感性增加7.43倍,逆转活性是VER的3.19倍,但对K562敏感株基本无影响。同时CH浓度依赖性地增加K562/ADR细胞内ADR和Rho123的蓄积,减少Rho123的泵出,抑制P糖蛋白的表达,但对K562细胞均无明显影响。CH在体外逆转肿瘤细胞多药耐药性的作用可能与其抑制P糖蛋白的功能和表达有关。In this study,cepharanthine hydrochloride（CH） was tested for its potential ability to modulate the expression and function of P-glycoprotein（P-gp） in the multidrug-resistant human chronic myelogenous leukemia cell line K562/ADR.Cytotoxicity of adriamycin（ADR） alone or in combination with CH or verapamil（VER） in K562 and K562/ADR cells was determined by MTT assay.Based on flow cytometric technology,the effect of CH or VER on the uptake and efflux of rhodamine123（Rho123） and the accumulation of ADR in these cells was detected by measuring Rho123 or ADR-associated mean fluorescence intensity（MFI）.The effects of CH and VER on P-glycoprotein（P-gp） expression in K562 and K562/ADR cells were also measured using a flow cytometry with PE-conjugated P-glycoprotein antibody.The results show that CH significantly enhanced the sensitivity of K562/ADR cells to ADR,4 μmol-L-1 of CH enhanced the sensitivity of K562/ADR cells to ADR by 7.43 folds,the reversal activity was 3.19 times higher than that of verapamil.However,CH had no effect on drug-sensitive K562 cells（P 0.05）.CH increased Rho123 and ADR accumulation in a concentration-dependent manner（2-8 μmol·L-1） and inhibited the efflux of Rho123 from these cells,but did not affect the accumulation and efflux of Rho123 from the wild-type drug-sensitive K562 cells.The inhibition effect of CH on P-gp expression in K562/ADR cells is in a time-and concentration-dependent manner.The reversal activity of CH is possibility related to inhibition of P-gp function and expression,which lead to an increased intracellular accumulation of anticancer drugs.

关 键 词：多药耐药性 P糖蛋白 盐酸千金藤碱 K562/ADR细胞 

分 类 号：R285.5[医药卫生—中药学]