wt1定量联合FCM在急性髓细胞白血病MRD监测中的临床应用

目的 探讨在急性髓细胞白血病(AML)中Wilms瘤基因1(wt1) mRNA定量联合多参数流式细胞术(FCM)分析监测微小残留病灶的临床应用.方法 采用实时荧光定量聚合酶链反应技术(qRT-PCR)检测35例AML患者的wt1表达水平;将患者根据不同亚型分组检测;并且对9例缓解和4例复发患者进行随访,检测wt1表达水平.采用FCM分析AML中微小残留病灶(MRD)水平.结果 与对照组比较,AML病例组wt1表达水平明显增高,差异有统计学意义(P<0.05).在初诊的各种AML亚型中,M2亚型wt1表达水平最高,M6亚型wt1表达水平最低.9 例随访患者提示在达到完全缓解时wt1表达水平下降,而4例随访复发患者在复发时再次升高.FCM检测MRD在不同阶段异常髓系细胞比例明显不同.联合qRT-PCR和FCM技术对MRD检测的敏感性和特异性大大提高,两者具有一致性;利用ROC曲线对复发病例进行分析得到的监测阈值为3.33%.结论 在急性髓细胞白血病中wt1 mRNA定量联合多参数流式细胞术分析可用于监测MRD、评估治疗效果、预后及预测疾病复发风险.

Clinical application of MRD in acute myelogenous leukemia bywt1 quantitative combined with FCM

Objective To discuss the clinical application of minimal residual disease in acute myelogenous leukemia(AML) by wt1 mRNA quantitative combined with multi-parameter flow cytometry (FCM).Methods Real time quantitative polymerase chain reaction (qRT-PCR) method was established for detecting wt1 gene expression level in 35 AML patients.The indexes were detected by different subtypes;And 9 cases of ease and 4 cases of recurrence in patients was followed-up and detected the wt1 level.The multiparameter flow cytometry was used to analyze the minimal residual disease in AML.Results The expression of wt1 gene was significantly higher than that of the control group.Significant difference was found(P<0.05).In the newly diagnosed AMLs, wt1 was the highest in M2 and the lowest in M6.Follow-up of 4 AML patients showed that wt1 gene expression level was markedly decreased after CR, but obviously increased after relapse.The proportion of abnormal myeloid cells in different phases significantly changed by FCM.There was no difference of minimal residual disease in AML by qRT-PCR and multiparameter flow cytometry.The ROC curve was used to analyze the recurrent cases to get the threshold value(3.33%).Conclusion The quantitative analysis of wt1 combined with multi-parameter flow cytometry can be used to monitor minimal residual disease in leukemia patients, assess the treatment efficacy and prognosis, and predict the risk of recurrence.