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目的:观察维甲酸(RA)在体外诱导HL60细胞(人早幼粒细胞白血病细胞株)凋亡的作用,探讨维甲酸治疗早幼粒细胞白血病的作用机制。方法:用DNA电泳、DNA片段定量测定技术、流式细胞术分析及光镜和电镜观察凋亡细胞。结果:在体外培养中加维甲酸50mg/L孵育4h,DNA片段率达557%±121%,而对照为125%±49%(P<0001,n=9);流式细胞术检测发现,在50mg/L维甲酸作用下,细胞凋亡达50%,对照为9%。电镜观察维甲酸组HL60细胞出现典型的凋亡细胞形态改变。维甲酸诱导HL60细胞凋亡具有剂量及时间依赖性,维甲酸作用6h,出现凋亡高峰;随维甲酸浓度升高,诱导凋亡作用明显增强。在环孢素A(CsA)存在时,低浓度的维甲酸也有明显诱导HL60细胞凋亡作用。结论:维甲酸在体外能诱导HL60细胞凋亡,CsA能增强维甲酸诱导HL60细胞凋亡活性。研究提示诱导白血病细胞凋亡是维甲酸治疗急性早幼粒细胞白血病的重要机制之一。
Objective: To observe the role of retinoic acid (RA) in inducing apoptosis of HL-60 cells (human promyelocytic leukemia cell line) in vitro and to explore the mechanism of retinoic acid in the treatment of promyelocytic leukemia. Methods: DNA electrophoresis, DNA fragment quantitative determination, flow cytometry analysis and light and electron microscopy of apoptotic cells. Results: In vitro incubation of retinoic acid at 50mg / L for 4h resulted in a DNA fragment rate of 55.7% ± 12.1% compared with 12.5% ± 4.9% (P <0001, n = 9). Flow cytometry showed that under the action of 50 mg / L retinoic acid, apoptosis reached 50% and the control was 9%. Electron microscope retinoic acid group HL 60 cells showed a typical morphological changes of apoptotic cells. Retinoic acid induced HL 60 cell apoptosis in a dose and time-dependent manner, retinoic acid for 6h, the peak of apoptosis appeared; With the increase of retinoic acid concentration, the induction of apoptosis was significantly enhanced. Cyclosporin A (CsA) in the presence of low concentrations of retinoic acid also significantly induced HL 60 apoptosis. Conclusion: Retinoic acid can induce HL-60 cell apoptosis in vitro, and CsA can enhance the activity of retinoic acid induced HL-60 cell apoptosis. Research suggests that induction of apoptosis in leukemia cells is one of the important mechanisms of retinoic acid in the treatment of acute promyelocytic leukemia.