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目的:研究5-氨基咪唑4-氨甲酰核糖核苷(AICAR)对人白血病HL-60细胞周期的影响及作用机制。方法:培养HL-60细胞,以CCK-8法行细胞增殖抑制实验,计算加入不同浓度AICAR(1 000μM、500μM、250μM、125μM)、不同时间点(24 h、48 h、72 h)对HL-60细胞增殖的抑制率,寻找最佳浓度和时间点,然后以流式细胞技术分析细胞周期,再以逆转录-PCR(RT-PCR)方法对细胞周期调控因子p21、p27、CDK2的表达进行半定量定性分析。结果:CCK-8实验显示AICAR浓度越大,HL-60细胞的增殖抑制率越高。流式细胞周期分析发现加入AICAR后细胞发生了S期停滞。RT-PCR结果显示,与对照组比较,加入AICAR后加药组HL-60细胞CDK-2的表达无明显变化,而p21、p27的表达明显增强。结论:AICAR可以通过上调细胞周期调控因子p21、p27的表达影响细胞周期,抑制HL-60细胞的增殖。
Objective: To study the effect of 5-aminoimidazole 4-carbamoyl-ribonucleoside (AICAR) on the cell cycle of human leukemia HL-60 cells and its mechanism. Methods: HL-60 cells were cultured and treated with different concentrations of AICAR (1 000μM, 500μM, 250μM, 125μM) at different time points (24 h, 48 h, 72 h) -60 cell proliferation, looking for the optimal concentration and time point, and then analyzed by flow cytometry cell cycle, and then by reverse transcription-PCR (RT-PCR) method of cell cycle regulators p21, p27, CDK2 expression Semi-quantitative qualitative analysis. Results: The results of CCK-8 showed that the greater the concentration of AICAR, the higher the inhibition rate of HL-60 cells. Flow cytometry analysis found that after the addition of AICAR cell S phase stagnation. The results of RT-PCR showed that compared with the control group, the expression of CDK-2 in HL-60 cells after adding AICAR did not change significantly, while the expressions of p21 and p27 were significantly increased. CONCLUSION: AICAR can affect the cell cycle and up-regulate the proliferation of HL-60 cells by up-regulating the expression of cell cycle regulators p21 and p27.