摘要
目的探讨细胞周期蛋白依赖性激酶(CDK)4/6对急性白血病细胞增殖及凋亡的影响。方法用CDK4/6抑制剂PD0332991 0.01、0.1、1、10和20μmol/L处理急性白血病细胞U937及RS4;11,24、48h后CCK-8法检测细胞增殖情况。U937细胞(实验U组)和RS4;11细胞(实验R组)分别加入PD0332991 1μmol/L和5μmol/L;另取正常细胞为对照组(对照U组、对照R组)。流式细胞术检测细胞凋亡及细胞周期,实时定量PCR检测Bax、Bcl-2、X染色体连锁凋亡抑制蛋白(XIAP)、CDK2、CDK6、Caspase-3mRNA表达。结果处理24h和48h后,U937和RS4;11细胞增殖率随药物浓度的增加而降低(P<0.05),且48h后细胞增殖率下降更明显(P<0.05)。与相应的对照组相比,实验U组和实验R组细胞凋亡率增加(P<0.05);G0/G1期细胞增加,而S期和G2/M期细胞减少(P<0.05);处理24h后Bax、Caspase-3mRNA表达升高,而Bcl-2、XIAP、CDK2、CDK6mRNA表达减少(P<0.05)。结论抑制CDK4/6的表达能减少急性白血病细胞增殖及促进细胞凋亡;其机制可能与Bax和Caspase-3表达上调以及Bcl-2、XIAP、CDK2、CDK6表达下调有关。
Objective To investigate the influence of cyclin-dependent kinase(CDK)4/6on the proliferation and apoptosis of acute leukemia cells.Methods The acute leukemia cells U937 and RS4;11were treated by the inhibitor of CDK4/6PD0332991 0.01,0.1,1,10 and 20μmol/L for 24 and 48hours.The cell proliferation was detected by CCK-8assay.The U937(group U)and RS4;11(group R)cells were treated by PD0332991 1μmol/L and 5μmol/L,respectively.Normal cells were taken as the controls(group CU and group CR).The cell apoptosis and cell cycle were examined by flow cytometry and the mRNA expressions of Bax,Bcl-2,X-linked inhibitor of apoptosis protein(XIAP),CDK2,CDK6 and Caspase-3were detected by RT-PCR.Results The rate of cell proliferation was decreased with the increase of the concentration of PD0332991(P<0.05),which was more obvious after 48 hours treatment than that after 24 hours treatment(P<0.05).Compared to groups of CU and CR,the rate of cell apoptosis and proportion of G0/G1 were increased,the proportion of S and G2/M was decreased,the mRNA expressions of Bax and Caspase-3were upregulated and those of Bcl-2,XIAP,CDK2 and CDK6 were downregulated after 24 hours treatment in groups of U and R(P<0.05).Conclusion The inhibition of CDK4/6can inhibit the proliferation and induce the apoptosis of acute leukemia cells,which may be related to the upregulation of Bax and Caspase-3and downregulation of Bcl-2,XIAP,CDK2 and CDK6.
引文
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