细胞调控因子SKP2的表达与HL-60/A耐药的关系

Relationship between Expression of Cell Regulatory Factor SKP2 and Drug Resistance of HL-60/A Cells

【目的】观察细胞周期调控因子SKP2、P27kip1在白血病耐药细胞株HL-60/A和非耐药细胞株HL-60细胞中的表达及细胞周期比例,探讨细胞增殖与耐药的关系。【方法】RT-PCR、Western blot检测SKP2、P27kip1、MRP mRNA及蛋白表达,流式细胞仪检测细胞周期,采用药物毒性分析方法检测细胞对化疗药物的敏感性。【结果】白血病耐药细胞株HL-60/A对阿霉素、柔红霉素、阿糖胞苷的耐药倍数分别为49.14倍、39.20倍和6.43倍;HL-60/A细胞MRP的表达高于HL-60;HL-60/A和HL-60细胞株G0/G1细胞期比例分别为(35.10±0.81)%和(43.96±1.12)%,两组比较P<0.05;HL-60/A细胞中SKP2的表达高于HL-60细胞;P27kip1的表达低于HL-60细胞。【结论】HL-60/A细胞多药耐药机制产生与MRP的表达增高有关。耐药HL-60/A处于增殖期的细胞比例高,具有更高的增殖活性。

[ Objective ] To observe the proportion of cell cycle and the expression of cell cycle regulators S-phase kinaseassociated protein 2 （SKP2） and P27^kipl in drug-resistant leukemia cell line HL-60/A and drug non-resistant cell line HL-60, and to investigate the relationship between proliferation and multi-drug resistance of leukemia cells. [Methods] The expression of SKP2, P27^kip1, and multi-drug resistance associated protein （MRP） mRNA were determined by RT-PCR and Western blot analysis. The cell cycle was analyzed by flow cytometry. Drug toxicity analysis was used to determine the sensitivity of the cells to chemotherapeutic drugs. [Results] Drug-resistant leukemia cell line HL-60/A showed 49.14-fold resistance to Adriamyein, 39.20- fold resistance to Daunorubicin, and 6.43-fold resistance to Arabinosylcytosine, respectively. MRP expression of HL-60/A cell was higher than that of HL-60 cell. The proportions of HL-60/A cells and HL-60 cells in the GO/G1 phase of the cell cycle were （35.10 ± 0.81）% and （43.96 ± 1.12）%, respectively （P 〈 0.05）. The expression of SKP2 was significantly higher in HL-60/A cells than that in HL-60 cells, but the expression of P27^kip1 in HL-60/A cells was lower. [ Conclusions ] The mechanism of multidrug resistance of HL-60/A cell line is related to up-regulated expression of MRP. The proportion of drug resistant cell line HL- 60/A at the proliferation stage is high which indicates that drug-resistant cells have higher proliferative activity.