急性早幼粒细胞患者治疗前后循环内皮细胞的变化及其影响因素的分析

Alteration of circulating endothelial cells from acute promyelocytic leukemia patients before and after treatment and its influential factors

目的：探讨急性早幼粒细胞白血病（acute promyelocytic leukemia,APL）患者治疗前后循环内皮细胞（circulating endothelial cells,CECs）数量、表面抗原及生物学特性的变化,并分析其与临床指标的关系。方法：免疫磁珠结合三色流式细胞仪分选计数APL患者初诊及治疗后CECs的数量;免疫荧光染色检测APL患者初诊及治疗后CECs表面CD146、CD31、CD144、VEGFR-2、CD45及CD133的表达水平。体外培养CECs进行血管生成实验及细胞集落形成率的测定。结果：APL患者CECs在外周血白细胞〉10×10^-9/L及CD34阳性患者中数量明显增多（P〈0.05）,高危、低危及中危3组患者的CECs数量也呈现明显的差异（P〈0.05）。32例APL患者诱导化疗后均获得完全缓解,治疗后CECs数量及其表面CD133表达水平均明显下降（P〈0.05）。同时治疗后APL患者CECs体外形成血管数目及细胞的集落形成率均明显减少（P〈0.05）。APL患者CECs数量治疗后/治疗前的比值与As_2O_3治疗1周后的尿砷浓度呈现明显负相关性（P〈0.05）。结论：精确计数并深入了解CECs生物学特性可能有助于评价APL的预后及设计治疗策略。

AIM：To determine the biological feature of circulating endothelial cells（ CECs） in acute promyelocytic leukemia（ APL） patients before and after treatment,and to analyze the relationship between CECs and the clinical characteristics.METHODS：The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cells were identified by immunofluorescence staining for the expression of CD146,CD31,CD144,VEGFR-2,CD45 and CD133.The CECs were cultured in vitro,and the tube formation and colony-forming rate were determined.RESULTS：Increased quantity of CECs was observed in CD34 positive group and group with WBC 10 × 10^-9/L（ P〈0.05）.The quantity of CECs had a significant difference among low risk,medium risk and high risk groups（ P〈0.05）.The positive rate of CD133 and quantity of CECs significantly reduced in 32 APL patients when they gain complete remission after treatment（ P〈0.05）.The amount of tube formation and colony-forming rate were significantly reduced after treatment（ P〈0.05）.The ratio of CECs quantity from APL patients after treatment to that before treatment had a negative correlation with arsenic concentration in urine on day 7 during As_2O_3treatment（ P〈0.05）.CONCLUSION：Accurately counting CECs may be helpful for evaluating prognosis and designing treatment strategy.