wt1定量联合FCM在急性髓细胞白血病MRD监测中的临床应用

Clinical application of MRD in acute myelogenous leukemia by wt1 quantitative combined with FCM

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。

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