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本研究主要探讨细胞酸化对罗丹明123(rhodamine Rh123)在不表达或低表达MDR1且分化程度较低的细胞中累积的影响,为抗白血病细胞多药耐药(multidrug resistance,MDR)寻找胞内酸化逆转的新方法。采用实时定量PCR技术检测MDR1基因在mRNA水平的表达;利用高钾缓冲液对细胞进行酸化,应用激光共聚焦显微镜测定HL-60、MSC及CD34+脐血细胞pHi值;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HL-60、MSC及脐血CD34+细胞内Rh123累积的影响。结果表明:细胞酸化3小时对HL-60、MSC及脐血CD34+细胞的活力无明显影响,细胞酸化能明显改变细胞对Rh123的累积;此外,细胞分化程度越低,其细胞内Rh123的累积越少。结论 :细胞酸化能逆转HL-60、MSC和脐血CD34+细胞的耐药。
This study was to investigate the effect of cell acidification on the accumulation of rhodamine 123 in non-expressing or low-expressing MDR1 cells with low degree of differentiation, and to search intracellularly for multidrug resistance (MDR) in leukemia cells. A new method of acidification reversal. Real-time quantitative PCR was used to detect the mRNA expression of MDR1. The cells were acidified with high potassium buffer. The pHi values of HL-60, MSC and CD34+ umbilical cord blood cells were measured by laser confocal microscopy. The cell acidification was observed by MTT assay. The influence of vitality; flow cytometry was used to detect the effect of cell acidification on the accumulation of Rh123 in HL-60, MSC and cord blood CD34+ cells. The results showed that the acidification of cells for 3 hours had no significant effect on the viability of HL-60, MSC and umbilical cord blood CD34+ cells. The acidification of cells significantly changed the accumulation of Rh123 cells. In addition, the lower the cell differentiation, the more the accumulation of Rh123 in the cells. less. Conclusion: Cell acidification can reverse the resistance of HL-60, MSC and cord blood CD34+ cells.