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目的研究小剂量阿糖胞苷和阿克拉霉素联合G-CSF(CAG方案)对HL-60细胞及其耐药株HL-60/A的作用机理。方法以白血病细胞株HL-60及其耐药株HL-60/A为实验模型,模拟临床给药方式,体外加入Ara-C、ACR、G-CSF三药,作用244、8小时后收集细胞,分别进行细胞计数,细胞形态观察,MTT法检测不同药物对两种细胞株的生长抑制率,流式细胞仪检测细胞早期凋亡标记AnnexinⅤ,细胞分化抗原CD11b以及进行细胞周期分析。结果经CAG作用48 h后,两种细胞株数量明显减少,形态显示凋亡小体增多;早期凋亡标记AnnexinⅤ明显增高,CD11b表达轻度升高,与CA组比较,结果有显著差异(P<0.05)。细胞周期分析显示,CAG作用24 h后,与CA组比较,S期细胞比例增高(P<0.05);48 h后比例下降,结果未见明显差异(P>0.05)。HL-60细胞与HL-60/A细胞比较,两种白血病细胞株经CAG作用48 h后,在细胞数量、细胞形态、凋亡细胞比例、CD11b表达及S期细胞比例的改变方面未见明显差异。结论CAG方案对白血病细胞株HL-60、HL-60/A的作用机制主要是通过G-CSF促使白血病细胞进入细胞周期S期,增加小剂量Ara-C、ACR对白血病细胞的细胞毒性,以诱导细胞凋亡为主,轻度诱导分化。
Objective To study the mechanism of low doses of cytarabine and aclacinomycin combined with G-CSF (CAG regimen) on HL-60 cells and their resistant strains HL-60 / A. Methods The leukemia cell line HL-60 and its drug-resistant strain HL-60 / A were used as experimental models to simulate clinical administration. Ara-C, ACR and G-CSF were added in vitro. The cells were collected after 8 and 8 hours The cell growth inhibition rates of two different cell lines were detected by MTT assay. Annexin V and CD11b were detected by flow cytometry and cell cycle analysis. Results After 48 h treatment with CAG, the numbers of the two cell lines were significantly reduced and the apoptotic bodies were increased. The early apoptotic marker AnnexinⅤ was significantly increased and the expression of CD11b was slightly increased (P <0.05). Cell cycle analysis showed that the percentage of cells in S phase increased after CAG treatment for 24 h compared with CA group (P <0.05), and decreased after 48 h (P> 0.05). Compared with HL-60 / A cells, the HL-60 / A cells showed no obvious changes in the number of cells, the cell morphology, the percentage of apoptotic cells, the expression of CD11b and the percentage of S phase cells after CAG treatment for 48 h difference. Conclusion The mechanism of action of CAG regimen on leukemia cell lines HL-60 and HL-60 / A is mainly through the G-CSF to promote leukemia cells into the cell cycle S phase and increase cytotoxicity of leukemia cells with low dose Ara-C, ACR Induced apoptosis primarily, slightly induced differentiation.