论文部分内容阅读
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 investigate the possibility and mechanism of multidrug resistance reversal by calmodulin antagonist berbamine (BBM). Methods MCF7 and ADR MCF7 ADR cells were co-cultured with ADR and different concentrations of BBM, and the IC50 values were calculated by MTT colorimetric assay. Fold. The relative content of ADR and the expression of P glycoprotein (P gp) were detected by flow cytometry. The mRNA expression of mdr1 was detected by semi-quantitative RT-PCR. Results The IC50 of ADR to MCF7 and MCF7 ADR were (98 ± 0 0 6) μmol L and (10 1 ± 0 ± 5 72) μmol L, respectively. The multidrug resistance of MCF7 ADR to ADR was 10 3. ADR cells sensitized with 5 μmol L, 10 μmol L and 20 μmol LBBM in a dose-dependent manner sensitized multiples of 2 76,5 88 and 2 8 2 6 (P <0.01), respectively ). MCF7 ADR cells were treated with 10 μmol or 20 μmol LBBM for 2 h, and the relative content of ADR in cells increased by 249 and 281-fold (P <0.01), and the expression of P gp decreased by 109% and 6 2 82% (all P <0 01), and the relative expression of mdr1 mRNA also showed a downward trend. Conclusion BBM can increase the intracellular ADR concentration and down-regulate the expression of mdr1 gene and P gp in MCF7 ADR cells, which makes it more sensitive to ADR.