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【目的】观察S期激酶相关蛋白2(SKP2)siRNA转染后耐阿霉素的HL-60细胞(HL-60/A)P27kip1及多药耐药相关蛋白MRP表达水平的变化,探讨其对细胞周期及药物敏感性的影响。【方法】siRNA转染HL-60/A细胞,荧光显微镜、流式细胞仪检测转染效率,RT-PCR、Western-blot检测转染后细胞SKP2、P27kip1、MRPmRNA及蛋白表达,流式细胞仪检测细胞周期,药物毒性分析CCK-8法检测细胞对化疗药物的敏感性。【结果】SKP2siRNA转染24h后,HL-60/A的SKP2mRNA及蛋白表达分别下调43%(P=0.020)和69.6%(P=0.003);P27kip1蛋白上调78.1%(P=0.049),P27kip1mRNA表达无明显变化;MRPmRNA及蛋白表达分别下调55%(P=0.001)和62.3%(P=0.001);G0/G1期细胞比例升高;细胞对阿霉素、柔红霉素、阿糖胞苷的敏感性分别提高1.15倍、4.79倍和3.76倍。【结论】SKP2siRNA下调HL-60/A细胞的SKP2表达可使P27kip1蛋白表达升高,细胞增殖周期延长;细胞耐药性的改善可能与MRP表达的降低有关。
【Objective】 To investigate the changes of P27kip1 and multidrug resistance-related protein MRP in HL-60 / HL-60 cells after transfection with S-phase kinase-related protein 2 (SKP2) siRNA Cell cycle and drug sensitivity. 【Methods】 The expression of SKP2, P27kip1, MRP mRNA and protein in transfected HL-60 / A cells was detected by fluorescence microscopy and flow cytometry. The expression of SKP2, P27kip1, MRP mRNA and protein were detected by RT-PCR and Western- Detection of cell cycle, drug toxicity analysis CCK-8 method to detect cell sensitivity to chemotherapy drugs. 【Results】 The results showed that the SKP2 mRNA and protein expression of HL-60 / A were down-regulated by 43% (P = 0.020) and 69.6% (P = 0.003) (P = 0.001) and 62.3% (P = 0.001) respectively. The percentage of cells in G0 / G1 phase increased. The cells were resistant to doxorubicin, daunorubicin, cytarabine The sensitivity increased by 1.15 times, 4.79 times and 3.76 times respectively. 【Conclusion】 The down-regulation of SKP2 expression by SKP2 siRNA in HL-60 / A cells can increase the expression of P27kip1 and prolong the cell cycle. The improvement of drug resistance may be related to the decrease of MRP expression.