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目的:研究屈洛昔芬(DRO)对耐阿霉素(ADR)K562细胞株(K562/A02)多药耐药性(MDR)的逆转作用及逆转机制。方法:用DRO分别处理K562/A02和K562敏感株。MTT法观察DRO影响K562/A02对ADR化学敏感性的变化。DRO 10μmol/L处理K562/A02前后,通过RT-PCR和免疫细胞化学染色,分析MDR1、GSTπ基因表达的变化,采用流式细胞技术测定细胞内ADR浓度的变化。结果:DRO显著逆转K562/A02的MDR,在20、10和5μmol/L浓度时,对ADR的化学敏感性分别增加到14、13和4倍,逆转活性与维拉帕米相当。MDR1和GSTπ的mRNA和蛋白表达在DRO 10μmol/L处理后第2天开始下降,第5天明显降低。用20、10和5μmol/L浓度的DRO处理两株细胞,K562/A02细胞内ADR积累分别增加到2.9、2.3和1.5倍。但DRO不能明显增加K562细胞内的ADR的浓度。结论:DRO对K562/A02的MDR有较强的逆转活性,逆转强度与维拉帕米相当,其逆转机制有多种不同的途径。
AIM: To investigate the reversal effect and reversal mechanism of droloxifene (DRO) on multidrug resistance (MDR) in adriamycin resistant K562 cell line (K562 / A02). Methods: K562 / A02 and K562 sensitive strains were treated with DRO respectively. The effect of DRO on the chemosensitivity of K562 / A02 to ADR was observed by MTT assay. The changes of MDR1 and GSTπ gene expression were analyzed by RT-PCR and immunocytochemistry before and after treatment with DRO 10 μmol / L K562 / A02. The changes of intracellular ADR concentration were determined by flow cytometry. RESULTS: DRO significantly reversed the MDR of K562 / A02 cells. The chemosensitivity to ADR increased to 14, 13 and 4 fold at 20, 10 and 5 μmol / L, respectively. The reversal activity was comparable to that of verapamil. The mRNA and protein expression of MDR1 and GSTπ began to decrease on the 2nd day after DRO 10μmol / L treatment, and decreased on the 5th day. Two cell lines were treated with DRO at concentrations of 20, 10 and 5 μmol / L. The accumulation of ADR in K562 / A02 cells increased to 2.9, 2.3 and 1.5 fold, respectively. However, DRO did not significantly increase the ADR concentration in K562 cells. CONCLUSION: DRO has a strong reversal activity on MDR of K562 / A02, and its reversal intensity is comparable to that of verapamil. There are many different ways of reversal mechanism.