【摘要】 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。更多还原

【Abstract】 Objective To investigate the significance of monitoring Wilms′ tumor gene （WT1） expression level in bone marrow of leukemia patients following allogeneic bone marrow transplantation （allo-BMT）.Methods Real-time quantitative reverse transcription polymerase chain reaction method was established for measuring WT1 and GAPDH expression levels in bone marrow cells of 15 patients with leukemia, including a total of 111 specimens during the follow-up, and in 23 non-leukemia patients by using LightCycler. Normalized WT1 expression level （WT1_N） was determined as a ratio of WT1 to GAPDH times 104.Results The median expression levels of WT1_N in 17 samples of newly diagnosed patients, 6 samples of relapsed patients, 88 samples from leukemia patient in complete remission and 23 samples of non-leukemic controls were 40.18 （5.48 to 510.27）, 125.89 （34.50 to 273.95）, 4.80 （0 to 56.96） and 1.47 （0 to 8.56） respectively. Nonparameter statistic analysis （Mann-Whitney U test） showed that the WT1_N expression levels in the newly diagnosed group and relapsed group were statistically higher than in those in the complete remission group and non-leukemic controls （all P<0.01）, without significant differences between the complete remission group and control group （P=0.692） and between the newly diagnosed group and relapsed group （P=0.595）. In general, the dynamic curves of WT1_N levels following allo-BMT were consistent with the tendency of changes in expression levels of corresponding fusion genes for minimal residual disease （MRD） monitoring. Spearman Rho correlation analysis revealed that the correlation coefficient between the WT1_N expression levels and BCR/ABL, AML/ETO, PML/ RARα and MLL/AF17 fusion genes expression were 0.678 （P=0.00）, 0.677 （P=0.00）, 0.806 （P=0.00） and 0.553 ^P=0.049 respectively. Three patients relapsed after allo-BMT and one patient relapsed before allo-BMT during the follow-up. A re-increment of WT1_N expression level during follow-up could be detected 40 to 180 days earlier to hematological relapse.Conclusion The WT1 expression level of leukemia patients following allo-BMT measured by real time RT-PCR can be a useful tool for monitoring MRD and warning the clinical relapse during follow-up.更多还原