目的探讨定量检测Wilms瘤基因1(WT-1)m RNA在急性白血病患者造血干细胞移植后的疗效、预后和预测复发中的价值。方法收集84例经异基因造血干细胞移植的急性白血病患者不同治疗时间的骨髓标本,实时荧光定量聚合酶链反应技术(RQ-PCR)分析WT-1 m RNA表达水平;聚合酶链反应-短串联重复序列法(PCR-STR)分析移植后供体基因嵌合率,并进行统计学分析。结果与AML和ALL治疗前组WT-1表达水平[分别为4.13(0.77,9.26)、0.69(0.16,2.35)]比较,AML和ALL治疗后组[分别为0.51(0.06,0.91)(U=164,P〈0.01);0.06(0.02,0.10)(U=315,P〈0.01]、缓解组[分别为0(0,0.06)(U=0,P〈0.01);0(0,0.03)(U=0,P〈0.01)]表达水平均明显下降。与缓解组相比,AML复发组[3.35(2.46,5.63)(U=0,P〈0.01)]、ALL复发组[2.46(2.17,3.31)(U=0,P〈0.01)]WT-1表达水平均明显上升。15例复发患者,在临床确诊复发前的WT-1水平明显升高。移植患者WT-1水平与供体基因嵌合率呈负相关(r=-0.73,P〈0.05)。结论 WT-1m RNA定量分析可用于监测经造血干细胞移植急性白血病患者的微小残留病灶,评估疗效、预后及预测疾病复发风险。
Objective To investigate the value of WT-1 m RNA quantitative analysis in monitoring the therapeutic effect,prognosis and relapse of the patients with hematopoietic stem cell transplantation for leukemia. Methods The bone marrow samples at different time from 84 patients with allogenic hematopoietic stem cell transplantation for leukemia were collected. The WT-1 m RNA levels in these samples were detected by real-time fluorescence quantitative PCR( RQ-PCR). The donor gene chimeric rate after transplantation was analyzed by the PCR-short tandem repetitive sequence( PCR-STR). Results The expression levels of WT-1 m RNA in the patients with initial AML( 4. 13 [0. 77,9. 26]) or ALL( 0. 69 [0. 16,2. 35]) were significantly higher than those after treatment( 0. 51[0. 06,0. 91],U = 164,P〈0. 01; 0. 06 [0. 02,0. 10],U = 315,P〈0. 01) or complete remission( CR)( 0 [0,0. 06],U = 0,P〈0. 01; 0 [0,0. 03],U = 0,P〈0. 01). Compared with the CR patients,the expression levels of WT-1 m RNA in the relapse AML( 3. 35 [2. 46,5. 63],U = 0,P〈0. 01) and ALL( 2. 46 [2. 17,3. 31],U = 0,P〈0. 01) patients increased significantly. The WT-1 m RNA levels increased two months before clinical relapse in 15 patients with recurrence. The WT-1 m RNA levels were negatively correlated with the donor gene chimeric rate( r =- 0. 73,P〈0. 05). Conclusion The quantitative analysis of WT-1 m RNA may be used to monitor the minimal residual disease of the leukemia patients after hematopoietic stem cell transplantation,and evaluate the patients' therapeutic effect,prognosis and relapse.