三氧化二砷对急性早幼粒细胞白血病细胞微粒数量及促凝活性的影响

摘要

Objective: To observe the effect of arsenic trioxide (ATO) on number, tissue factor (TF) expression and procoagulant activity of microparticles released by acute promyelocytic leukemia (APL) cells, and to identify how much TF plays a role in procoagulant activity of these microparticles. Methods: APL cells were isolated from bone marrow of 3 newly diagnosed APL patients. Bone marrow mononuclear cells were collected from 3 patients with iron deficiency anemia and used as control. APL cells were cultured with medium containing different concentration of ATO for 24 h, 48h or 72h. Microparticles were extracted from the cell culture medium. The number and TF expression of microparticles were measured by flow cytometry. The procoagulant activities of microparticles released by different groups of cells were detected by coagulation assays. How much TF plays a role in the procoagulant activity of APL cells-derived microparticles (APL-MPs) was evaluated by coagulation inhibition assays in which anti-TF antibody was used to suppress TF procoagulant activity. Results: 1.0 μM and 2.0 μM ATO had a promoting effect on the release of microparticles by APL cells. Compared with bone marrow mononuclear cells-derived microparticles, the bone marrow APL-MPs showed a higher TF expression and procoagulant activity. 0.5 μM and 1.0 μM ATO had an inhibitory effect on TF expression and procoagulant activity of APL-MPs in a time-dependent manner. After preincubation with anti-TF antibody, in most groups, the APL-MPs indicated a significantly decreased procoagulant activity (prolonged clotting time). Conclusion: Both the TF expression and procoagulant activity of APL-MP were obviously increased, and TF played an important role in the procoagulant activity of these microparticles. ATO could promote the release of microparticles by APL cells. Low concentration of ATO could effectively reduce the TF expression and procoagulant activity of APL-MP.%目的:观察急性早幼粒细胞白血病(APL)细胞来源微粒(APL-MP)的促凝活性、表面组织因子(TF)表达情况、TF在其促凝活性中发挥的作用及分化治疗药物三氧化二砷(ATO)对上述指标有何影响.方法:选取3例初发APL患者,提取骨髓APL细胞,3名缺铁性贫血患者提取骨髓单个核细胞作为对照.分别用不同浓度ATO处理APL细胞24h、48h、72 h,收集细胞培养液提取微粒.采用流式细胞术对微粒进行定量分析并进行微粒表面TF表达情况检测；利用凝血实验比较不同组细胞释放微粒的促凝血活性；应用抗TF抗体抑制微粒促凝血活性实验检测TF在APL-MP的促凝血活性中发挥多大作用.结果:1.0 μM及2.0 μM ATO能显著促进APL细胞释放微粒.与正常骨髓来源单个核细胞释放的微粒相比,骨髓APL-MP的TF表达及促凝活性均显著增高,0.5μM及1.0 μM ATO处理可以有效降低APL-MP的TF表达及促凝活性,且这一作用呈时间依赖性.各组APL-MP经抗TF抗体孵育后凝血时间显著延长.结论:APL-MP的TF表达和促凝学活性均显著增高,并且TF在APL-MP的促凝血活性中发挥着重要作用.ATO能显著促进APL细胞释放微粒,低浓度ATO可以有效降低APL-MP的TF表达及促凝血活性.