全仅式维甲酸(ATRA)和三氧化二砷(As2O3)对APL细胞组织因子表达的影响

摘要

Objective　To study the effect of all-trans retinoic acid (ATRA) and arsenic trioxide (As2O3) on tissue factor (TF) expression and procoagulant activity (PCA) of acute promyelocytic leukemia (APL) cells in vivo and in vitro. rnMethods　PCA from freshly isolated APL blasts from APL patients treated with ATRA or As2O3 was detected using a one-stage clotting assay. TF antigen was detected by ELISA and TF mRNA by RT-PCR. The maturation sensitive (NB4) or resistant subclones (NB4-R1) of the promyelocytic NB4 cell line, as well as U937 cells infected with pMSCV-PML-RARa treated with or without ATRA or As2O3, were also examined. rnResults　Both ATRA and As2O3 can down-regulate the TF antigen, its mRNA transcription and membrane PCA of APL cells in vivo and in vitro, in a time-dependent manner. The TF antigen level in PML-RARa+ U937 cells was significantly higher than that in U937 cells infected with retrovirus vector. Both ATRA and As2O3 can also down-regulate the TF antigen in U937 cells transfected with or without PML-RARa. rnConclusion　Tissue factor expression and PCA in APL cells may be down-regulated by ATRA and As2O3. By down-regulating TF expression, As2O3 might also be used to improve the DIC-related hemorrhage in APL. Our data indicate that elevated TF antigen in PML-RARa+ U937 may be related to the fusion protein PML-RARa. The down-regulating effect of ATRA and As2O3 on TF expression in U937 cells might not involve this fusion protein.%目的探讨全反式维甲酸（ATRA）和三氧化二砷（As2O3）在体内外对急性早幼粒细胞性白血病（APL）细胞组织因子（TF）表达的影响。rn方法利用复钙时间测定、ELISA和RT-PCR等方法，分别检测了ATRA和As2O3治疗前后APL患者骨髓单个核细胞的促凝活性、TF抗原及其mRNA的转录水平，同时还检测了使用ATRA和As2O3处理APL细胞株NB4细胞和NB4-R1细胞以及转染PML-RARa融合基因的U937细胞的促凝活性、TF抗原及其mRNA的转录水平。rn结果 ATRA和As2O3在体内和体外均可以时间依赖的方式下调NB4细胞的促凝活性、TF抗原水平以及TF mRNA的转录。转染PML-RARa融合基因的U937细胞与仅转染逆病毒载体的U937细胞相比，其TF表达水平显著升高；使用ATRA或As2O3分别处理转染PML-RARa和转染逆病毒载体的U937细胞，其TF抗原水平均显著降低。rn结论 ATRA和As2O3均可下调APL细胞TF的表达并降低其PCA，As2O3在诱导APL细胞凋亡的同时有可能通过下调APL细胞TF的表达而改善APL患者与DIC相关的出血症状。结果还提示APL细胞染色体易位产生的融合蛋白PML-RARa可能对TF的异常表达有一定的影响，而ATRA和As2O3对TF的下调作用可能不依赖于对融合蛋白PML-RARa的降解。