急性早幼粒细胞白血病细胞来源微粒在凝血紊乱中的作用研究

摘要

目的:评估急性早幼粒细胞白血病(acute promyelocytic leukemia,APL)细胞来源微粒(microparticle,MP)及其携带的组织因子(tissue factor,TF)在患者高凝状态中的作用,以及化疗/分化治疗药物对MP促凝血活性(pro-coagulant activity,PCA)的影响.方法:提取5例APL患者和年龄性别相匹配的5例缺铁性贫血患者(对照)的骨髓单个核细胞(MNC),体外培养48 h,收集含MP细胞培养液和去MP细胞培养液,从一定体积的含MP细胞培养液中提取MP.采用ELISA法检测MP上TF的表达,应用凝血酶生成实验检测含MP细胞培养液和去MP细胞培养液的PCA,利用人TF特异抗体检测TF在MP相关PCA中的作用.测定全反式维甲酸(ATRA)、三氧化二砷(ATO)和柔红霉素(DNR)处理APL细胞48 h后MP的PCA.结果:对照者骨髓MNC释放的MP几乎无TF表达,而APL细胞来源的MP明显表达TF.无论是APL患者骨髓MNC还是对照者骨髓MNC,释放的MP均具有明显的PCA,但与对照者骨髓MNC释放的MP相比,APL患者骨髓MNC释放的MP的PCA更高.TF在对照者骨髓MNC释放的MP的PCA中几乎无作用,而在APL患者骨髓MNC细胞来源的MP的PCA中发挥重要作用.DNR处理的APL患者骨髓MNC能使其释放的MP的PCA增高,而ATO和ATRA的作用则完全相反.结论:APL患者骨髓MNC来源的微粒具有明显的PCA,而TF在这一活性中发挥着重要作用.化疗药物DNR可增强APL患者骨髓MNC释放的MP的PCA明显,而分化治疗药物ATRA和ATO则可降低这一活性.%Objective:To evaluate the role of microparticle (MP) derived from acute promyelocytic leukemia (APL) cells and tissue factor (TF) carried by the MP in hypercoagulable state,and the effect of treatment with cytotoxic chemotherapy/differentiating agents on procoagulant activity (PCA) of these MP.Methods:Bone marrow mononuclear cells (BMMNC) were extracted from 5 APL patients and 5 sex-and age-matched patients with iron deficiency anemia as controls.The cells were cultured in vitro for 48 h,then MP-rich culture medium and MP-free culture medium were harvested and MP was further obtained from certain volume of MP-rich culture medium.Subsequently,TF expression on MP was measured by ELISA.PCA of MP-rich culture medium or MP-free culture medium was assessed with thrombin generation assay.The role of TF on MP-related PCA was evaluated using antihuman TF antibody.In addition,APL cells were treated with all-trans retinoic acid (ATRA),arsenic trioxide (ATO) or daunorubicin (DNR) for 48 h,then MP-rich culture medium were harvested and the PCA was determined.Results:No TF expression was found in the MP released from bone marrow MNC in control group,whereas the obvious TF expression was found in the MP originated from BMMNC of APL.MP from both APL and control BM-MNC had obvious PCA.However,compared with the MP derived from control MNC,the MP from APL BM-MNC induced significantly higher PCA.TF played a crucial role in the PCA of APL BM-MNC derived MP,while played no role in that of MP from control MNCs.DNR-treating APL BM-MNC resulted in an increase in the PCA of MP,whereas ATO or ATRA exposure lead to exactly the opposite results.Conclusion:MP derived from APL BM-MNC posseses obvious PCA.TF plays a crucial role in the MP-related PCA.The PCA of MP increases after treating APL BM-MNC with chemotherapy agent DNR and decreases following exposure of APL BM-MNC to differentiating agents ATRA or ATO.