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目的探讨cariporide对耐药细胞株K562/DOX中P-糖蛋白(P-glycoprotein,P-gp)的影响,为抗白血病多药耐药(multi-drugresistance,MDR)提供新方法。方法应用cariporide对细胞进行酸化,应用激光共聚焦显微镜测定野生型细胞系K562及耐药细胞株K562/DOX细胞内pH值及细胞酸化对K562和K562/DOX细胞内阿霉素累积的影响。采用MTT法观察细胞酸化对细胞活力的影响。应用流式细胞术检测细胞酸化对K562/DOX细胞中P-gp功能的影响。采用实时定量RT-PCR技术检测MDR1基因在mRNA表达水平的变化。结果 Cariporide处理3h对K562及K562/DOX细胞的活力影响较小。在K562/DOX细胞中,P-gp的外排药物能力随细胞内pH值的降低而减弱,cariporide明显增加了细胞对罗丹明123(rhodaminel123,Rh123)和阿霉素的累积。细胞酸化还在mRNA水平抑制了K562/DOX细胞中P-gp的表达。结论 Cariporide能够抑制K562/DOX耐药细胞株中MDR1基因表达和P-gp的功能。
Objective To investigate the effect of cariporide on P-glycoprotein (P-gp) in K562 / DOX cell line and to provide a new method for anti-leukemia multi-drug resistance (MDR). Methods The cells were acidified with cariporide. The effect of intracellular pH and cell acidification on the accumulation of doxorubicin in K562 and K562 / DOX cells was detected by laser confocal microscopy. The effects of cell acidification on cell viability were observed by MTT assay. The effect of cell acidification on the function of P-gp in K562 / DOX cells was detected by flow cytometry. Real-time quantitative RT-PCR was used to detect the mRNA expression of MDR1. Results Cariporide treatment 3h had little effect on the viability of K562 and K562 / DOX cells. In K562 / DOX cells, the ability of efflux drug of P-gp decreased with the decrease of intracellular pH value. Cariporide significantly increased the accumulation of rhodamine123 (Rh123) and doxorubicin in cells. Cell acidification also inhibits P-gp expression in K562 / DOX cells at the mRNA level. Conclusion Cariporide can inhibit the MDR1 gene expression and the function of P-gp in K562 / DOX resistant cell lines.