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目的:探讨脉冲磁场(pulsed magnetic fields,PMF)对白血病细胞HL60/ADR多药耐药的逆转效果及其可能的分子机制。方法:生长曲线法检测PMF的细胞毒性;MTT法测定HL60、HL60/ADR的IC50及经不同参数PMF作用后HL60/ADR的IC50,计算耐药倍数和逆转倍数;流式细胞术检测PMF作用前后HL60/ADR细胞内药物蓄积变化及多药耐药相关蛋白(Multidrug Resis-tance Related Protein1 MRP1)阳性表达率的变化;RT-PCR法检测MRP1基因表达的变化。结果:单独PMF对HL60/ADR细胞的生长没有明显抑制作用,不同参数(频率为150Hz和250Hz,场强均为40mT,照射时间30min和1h)的PMF均能有效逆转HL60/ADR的多药耐药,150Hz/1h作用最为明显,逆转倍数为5.891倍。PMF作用后Rh123在HL60/ADR细胞内的蓄积增加了近8.0%,而MRP1基因表达下调了68.3%,蛋白阳性表达率下调23.6%。结论:PMF能够通过下调MRP1基因和蛋白的表达,进而增加Rh123在细胞内的蓄积,功能性部分逆转白血病细胞的多药耐药。
Objective: To investigate the reversal effect of pulsed magnetic fields (PMF) on multidrug resistance of HL60 / ADR leukemia cells and its possible molecular mechanism. Methods: The cytotoxicity of PMF was detected by the growth curve method. The IC50 of HL60, HL60 / ADR and the IC50 of HL60 / ADR after different parameters of PMF were determined by MTT method. The antimicrobial resistance and reversal fold were calculated by flow cytometry The change of drug accumulation in HL60 / ADR cells and the expression of Multidrug Resis-tance Related Protein 1 (MRP1) were detected by RT-PCR. The changes of MRP1 gene expression were detected by RT-PCR. RESULTS: PMF alone had no significant inhibitory effect on the growth of HL60 / ADR cells. PMFs with different parameters (frequency of 150 and 250 Hz, field intensity of 40 mT and irradiation time of 30 min and 1 h) all could effectively reverse the multidrug resistance of HL60 / ADR Medicine, 150Hz / 1h most obvious role, reversal fold 5.891 times. The accumulation of Rh123 in HL60 / ADR cells increased by nearly 8.0% after PMF treatment, while the expression of MRP1 gene was down-regulated by 68.3% and the protein positive expression rate was reduced by 23.6%. Conclusion: PMF can down-regulate the expression of MRP1 gene and protein, thereby increasing the accumulation of Rh123 in cells and partially reversing the multidrug resistance of leukemic cells.