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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 effect of cyclin dependent kinase (CDK) 4/6 on the proliferation and apoptosis of acute leukemia cells. Methods Acute leukemia cells U937 and RS4 were treated with CDK4 / 6 inhibitor PD0332991 at 0.01, 0.1, 1, 10 and 20 μmol / L, respectively. Cell proliferation was measured by CCK-8 at 11, 24 and 48 hours after treatment. U937 cells (experimental group U) and RS4; 11 cells (experimental group R) were added PD0332991 1μmol / L and 5μmol / L respectively; other normal cells were control group (control U group, control group R). The apoptosis and cell cycle were detected by flow cytometry. The expression of Bax, Bcl-2 and X-linked inhibitor of apoptosis protein (XIAP), CDK2, CDK6 and Caspase-3 mRNA were detected by real-time quantitative PCR. Results After treatment for 24h and 48h, the proliferation rate of U937 and RS4; 11 cells decreased with the increase of drug concentration (P <0.05), and the cell proliferation rate decreased more significantly after 48h (P <0.05). Compared with the corresponding control group, the apoptosis rate of experimental U group and experimental R group increased (P <0.05), while the cells in G0 / G1 phase increased, while the cells in S phase and G2 / M phase decreased (P <0.05) After 24h, the expression of Bax and Caspase-3 mRNA increased, while the expression of Bcl-2, XIAP, CDK2 and CDK6 decreased (P <0.05). Conclusions Inhibition of CDK4 / 6 expression can reduce the proliferation and promote the apoptosis of acute leukemia cells. The mechanism may be related to the up-regulation of Bax and Caspase-3 and the down-regulation of Bcl-2, XIAP, CDK2 and CDK6.