亚砷酸诱导分化的急性早幼粒细胞白血病细胞浸润人脑膜组织的体外模拟及其分子机制研究

目的:探讨急性早幼粒细胞白血病(Acute promyelocytic leukemia,APL)合并中枢神经系统白血病的发病机制。方法:采用流式细胞术检测亚砷酸(Arsenious acid, ATO)诱导分化前后的APL细胞及人APL细胞株NB4细胞表面CD56、CXCR4的表达;用荧光染料-羧基荧光素二醋酸盐琥珀酰亚胺酯标记ATO分化的APL(APL/ATO)、NB4细胞(NB4/ATO);用微重力旋转培养法体外模拟APL细胞浸润人脑膜组织,观察组织学及超微结构。结果:ATO诱导后,APL/ATO细胞表面CXCR4的表达明显高于诱导前(35.2±9.5%vs. 18.6±4.9%);NB4/ATO细胞表面CXCR4的表达明显高于诱导前(39.6±2.6%vs. 21.0±7.3%);APL/ATO细胞表面CD56的表达明显高于诱导前(36.6±8.9%vs. 25.8±5.15%);NB4/ATO细胞表面CD56的表达明显高于诱导前(44.6±8.4%vs. 25.6±2.4%)。组织学实验结果显示对照组脑膜组织未见NB4、APL细胞浸润,实验组可见APL/ATO、NB4/ATO细胞浸润到人脑膜组织中;荧光显微镜下可见被标记的APL/ATO、NB4/ATO细胞浸润到人脑膜组织中,扫描电镜见APL/ATO、NB4/ATO细胞浸润到脑膜组织中。结论:本研究采用微重力旋转培养系统体外模拟了ATO诱导分化的异常早幼粒细胞浸润人脑膜组织,APL细胞和NB4细胞CXCR4、CD56的表达升高可能是ATO诱导治疗APL所致的中枢神经系统浸润的分子机制之一。

In Vitro Simulation Experiment of APL Specialized by Arsenic Trioxide Acid infiltra Ting into the Human Meninges and its Molecular Mechanism

ABSTRACT Objective: To explore the pathogenesis and preventive measure of acute promyelocytic leukemia complicated with central nervous system leukemia. Methods: The expression of CXCR4 and CD56 on differentiated APL were detected by using flow cytometry (FCM). The differentiated APL and NB4 were labeled by 5, 6-car-boxyfluorescein diacetate succinimidyl ester(CFSE); Rotary cell culture system (RCCS) was used to model APL infiltration meninges to observe the histology and ultramicrostructure in vitro. Results: The average expression rate of CXCR4 on the ATO differentiated APL were 35.2 ± 9.5 %, which was significantly higher than that before induction(18.6 ±4.9 %). The average expression rate of CXCR4 on the ATO differentiated NB4 was 39.6 ± 2.6 %, which was significantly higher than that before induction(21.0 ±7.3 %). The average expression rate of CD56 on the ATO differentiated APL was 36.6 ± 8.9 %, which was significantly higher than that before induction(25.8 ±5.15 %). The average expression rate of CD56 on the ATO differentiated NB4 was 44.6 ± 8.4 %, which was significantly higher than that before induction(25.6 ±2.4 %). The differentiated APL and NB4 infiltrated into meninges was observed in the trial groups but it can not be observed in the control group. Under the fluorescence microscope, the differentiated APL and NB4 were labeled by CFSE infiltrated into meninges in the trial groups. Under the electron microscope observation, the differentiated APL and NB4 infiltrated into meninges in the trial groups. Conclusion: The expression of CXCR4 and CD56 were increased after ATO induced differentiation of APL and NB4 cells. The abnormal differentiated APL and NB4 can infiltrate human meninges using RCCS in vitro, which provide a cytology basis for approaching pathogenesy of central nervous system infiltration in treatment of APL using ATO.