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目的探讨白血病患者接受异基因骨髓移植(allo BMT)后骨髓中WT1的动态表达水平在监测微小残留病(MRD)中的意义。方法建立实时定量逆转录聚合酶链反应(RT PCR)方法,采用LightCyclerPCR仪检测了15例白血病患者allo BMT前后(111例次)和23例非白血病患者骨髓中WT1及内参GAPDH的表达水平,以WT1N=(WT1拷贝数/GAPDH拷贝数)×104计算WT1表达水平。结果初诊与复发白血病患者骨髓中WT1N的中位表达水平分别为40 18和125 89,明显高于完全缓解组和对照组(分别为4 80和1 47),对照组与完全缓解组之间及初诊与复发组之间差异无统计学意义;动态检测WT1N变化与相应融合基因变化具有很好的相关性, WT1N表达水平再次升高可提前40~180d预示白血病临床复发。结论实时定量RT PCR动态检测WT1在白血病患者allo BMT后骨髓中表达水平,可作为监测残留病的指标,提示复发。
Objective To investigate the significance of the dynamic expression of WT1 in bone marrow of leukemia patients after allogeneic bone marrow transplantation (allo BMT) in the monitoring of minimal residual disease (MRD). Methods Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of WT1 and GAPDH in the bone marrow of 15 leukemia patients before and after allo BMT (111 cases) and 23 non-leukemia patients with LightCycler PCR WT1N = (WT1 copy number / GAPDH copy number) × 104 The WT1 expression level was calculated. Results The median expression of WT1N in the newly diagnosed and relapsed leukemia patients was 40 18 and 125 89 respectively, which was significantly higher than that of the complete remission group and the control group (4 80 and 1 47, respectively), and between the control group and the complete remission group There was no significant difference between the newly diagnosed group and the recurrent group. The dynamic changes of WT1N showed a good correlation with the changes of the corresponding fusion genes. The increased expression of WT1N could predict the recurrence of leukemia 40 ~ 180 days earlier. Conclusion Real-time quantitative RT-PCR can detect the expression of WT1 in the bone marrow of patients with leukemia after allo BMT, which can be used as an indicator of residual disease.