摘要
目的:观察孕酮和孕酮受体拮抗剂米非司酮(RU486)对K562白血病细胞增殖和分化的影响。方法:用10-5 mol/L孚酮分别联合10-5 mol/L、10-6 mol/L和10-7 mol/L RU486共同作用K562白血病细胞5 d,采用液体培养,XTT实验和集落培养实验观察K562细胞增殖能力;用流式细胞术检测K562细胞周期;通过Wright-Giemsa染色、联苯胺染色和表面分化抗原CD71表达观察孕酮和RU486对K562细胞分化的作用。结果:液体培养,XTT实验和集落形成抑制实验均显示单独使用孕酮可显著抑制K562细胞增殖,与对照组比较有显著差异(P<0.01)。孕酮联合RU486对K562细胞的增殖抑制作用减弱,与单独使用孕酮组比较有显著差异(P<0.01),呈剂量依赖关系。形态学观察孕酮处理组K562细胞体积变大,核浆比例减少,趋向成熟分化。联苯胺染色吸光度值升高,与对照组比较差异显著(P<0.01)。流式细胞术检测10-5 mol/L孕酮处理4 d后K562细胞膜上CD71表面标记表达阳性率为(85.72±2.31)%,显著高于对照组(P<0.01);但孕酮联合RU486对K562细胞的诱导分化作用减弱,与单独使用孕酮组比较有显著差异(P<0.01),并呈剂量依赖关系。细胞周期显示孕酮可将K562细胞周期阻滞在S期。但孕酮联合RU486组分布于S期的K562细胞百分比显著下降,与单独使用孕酮组比较差异显著(P<0.01),并呈剂量依赖关系。结论:孕酮可抑制K562细胞增殖并诱导其向红系分化,而RU486可阻断P对K562细胞的增殖抑制与诱导分化作用。
AIM：To investigate the effects of progesterone and progesterone receptor antagonist mifepristone on human leukemia cell line K562.METHODS： The K562 cells were treated with 10-5 mol/L progesterone alone or combined with mifepristone at the concentrations of 10-5,10-6 and 10-7 mol/L in the culture.The proliferative effects of progesterone and mifepristone on K562 cells were studied by liquid culture,colony count and XTT assay,and the differentiation effects were determined by morphological observation,benzidine staining and flow cytometry analysis.RESULTS： The growth of K562 cells was inhibited by progesterone treatment for 5 days in liquid culture,colony count and XTT assay（P0.01）.Compared with progesterone alone group,the combination of progesterone with mifepristone at different concentrations attenuated the inhibition of cell growth（P0.01）.Progesterone also induced erythroid differentiation in K562 cells,with bigger cell volume,decreased karyoplasmic ratio and ripening of differentiation.After progesterone treatment,the A value of bezindine staining was higher than that in control group（P0.01）.After progesterone treatment for 4 days,the percentage of CD71 expression in differentiated K562 cells was（85.72±2.31）%.Compared with progesterone alone group,the A value of bezindine staining and the percentage of CD71 expression in differentiated K562 cells were decreased by the combination of progesterone with mifepristone at different concentrations（P0.01）.The cell cycle of K562 cells was arrested in S-phase by progesterone treatment,and the number of the cells in S-phase was decreased by the combination of progesterone with mifepristone at different concentrations（P0.01）.CONCLUSION： Progesterone inhibits the proliferation of K562 human leukemia cells and induces their erythriod differentiation.Progesterone receptor antagonist mifepristone partly blocks the anti-proliferation and differentiation-inducing effects of progesterone.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2011年第6期1097-1102,共6页
Chinese Journal of Pathophysiology
基金
河南省教育厅自然科学研究计划资助项目(No.2010B320011)
关键词
孕酮
K562细胞
细胞增殖
细胞分化
Progesterone
K562 cells
Cell proliferation
Cell differentiation
