摘要
目的 探讨钙调素拮抗剂小檗胺 (BBM)逆转多药耐药的可能性及其机制。 方法 乳腺癌细胞MCF7及其耐阿霉素 (ADR)MCF7 ADR细胞与ADR及不同浓度BBM共培养 ,经四唑盐 (MTT)比色法计算半数抑制浓度 (IC50 )、耐药倍数和增敏倍数 ;流式细胞仪检测细胞内ADR相对含量和P糖蛋白 (P gp)表达水平 ;半定量RT PCR检测mdr1基因mRNA表达。 结果 ADR对MCF7和MCF7 ADR的IC50 分别为 (0 98± 0 0 6 ) μmol L和 (10 1 2 0±5 72 ) μmol L ;MCF7 ADR对ADR的耐药倍数为 10 3。MCF7 ADR细胞 5 μmol L、10 μmol L和 2 0 μmol LBBM时对ADR呈剂量依赖性增敏作用 ,增敏倍数分别为 2 76、5 88和 2 8 2 6倍 (均P值 <0 0 1)。用 10 μmol 或 2 0 μmol LBBM处理MCF7 ADR细胞 2h ,细胞内的ADR相对含量增加 2 4 9和 2 81倍 (P <0 0 1) ;处理 72h使P gp表达分别下降10 0 9%和 6 2 82 % (均P值 <0 0 1) ,且mdr1基因mRNA相对表达值也呈下调趋势。 结论 BBM提高耐药细胞MCF7 ADR胞内的ADR浓度 ,下调mdr1基因及P gp的表达 ,使其对ADR的敏感性显著增高。
Objective To explore the efficacy of calmodulin antagonist berbamine(BBM)on multidrug resistance(MDR)reversal and its mechanism. Methods Human breast cancer cell line MCF7 and its adriamycin-resistant counterpart MCF7/ADR were used in the study.The cells were cultured with ADR and different concentration of BBM. MTT assay was used to analyze the effect of BBM on cell growth inhibition.According to the MTT assay,the 50% inhibitory concentration(IC 50 ),the multiples of drug resistance and increased sensitivity of ADR were calculated.The concentration of intracellular ADR and expression level of P-glycoprotein(P-gp)were detected by flowcytometry(FCM).The mRNA expression level of mdr1 gene was detected by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR)with β-actin as internal reference. Results The IC 50 of ADR in MCF7 and MCF7/ADR cells were(0.98±0.06)μmol/L and(101.20±5.72)μmol/L,respectively.The resistant multiple of MCF/ADR cells to ADR was 103 folds higher than that of MCF7 cells.BBM increased the chemo-sensitivity of ADR in MCF7/ADR cells with dose-dependent relationship,i.e.when 5*!μmol/L ,10*!μmol/L and 20*!μmol/L BBM was added into the culture the chemo-sensitivity of ADR was increased to 2.76,5.88,and 28.26 folds(P<0.01),respectively.After treating MCF7/ADR cells by 10*!μmol/L or 20*!μmol/L BBM for 2 hours the accumulation of intracellular ADR was increased to 2.49 and 2.81 folds(P<0.01),while treating for 72 hours the expression of P-gp was decreased 10.09% and 62.82%(P<0.01),respectively,with down-regulated mRNA expression of mdr1 gene.Conclusion BBM could increase intracellular concentration of ADR in MCF7/ADR,down-regulate expre-ssion level of both mdr1 mRNA and P-gp so that the sensitivity of MCF7/ADR to ADR was increased significantly.
出处
《解剖学报》
CAS
CSCD
北大核心
2004年第2期161-164,共4页
Acta Anatomica Sinica
基金
美国中华医学基金会 (CMB)资助项目
