实时荧光定量PCR检测白血病患者WT1基因表达及临床意义

目的 研究白血病患者WT1基因的表达水平及与临床预后的关系.方法 建立实时荧光定量PCR检测WT1基因表达技术,并分析65例白血病患者WT1基因的表达水平及与临床预后的关系.结果 建立的实时荧光定量PCR方法的标准曲线相关系数＞0.99.急性髓性白血病(AML)和急性淋巴细胞白血病(ALL)患者WT1基因表达水平较正常对照组均显著升高(P=0.001).慢性髓性白血病(CML)急变期患者WT1基因表达水平显著高于慢性期.WT1表达量与急性白血病发病时外周血白细胞计数、血红蛋白、血小板无明显相关关系.AML和ALL患者中WT1基因高表达患者与低表达病例CR率差异无显著性.急性白血病患者治疗缓解后WT1的表达量较治疗前显著下降(P=0.032).随访显示WT1持续高表达或下降后又上升的患者呈现难治及复发.结论 实时荧光定量PCR技术检测急性白血病患者WT1基因表达量可预测难治复发及用于MRD的检测.

Clinical significance of real - time fluorescent quantitative PCR for detection of WT1 gene expression in leukemia patients

Objective To evaluate the relationship between the quantification of WT1 mRNA and prognosis in patients with leukemia.Methods WT1 mRNA level was detected with real time fluorescent quantitative PCR (RT-FQ-PCR) technique in bone marrow samples from 65 leukemia patients.Results The correlation coefficiency was over 0. 99 for standard curves of RT-FQ-PCR method. The median expression levels of WT1 in 43 newly diagnosed acute myeloid leukemia(AML) and 15 newly diagnosed acute lymphoblastic leukemia(ANLL) patient were significantly higher than those in normal control group(P=0.001). The WT1 expression level of chronic myeloid leukemia(CML) in blast crisis was higher than that of CML in chronic phase. The WT1 level was not correlated to peripheral WBC counts, hemoglobin and platelet counts. No significant differences of CR rate was found between the group with high WT1 expression levels and the group with low WT1 expression levels both in AML and ALL patients. The WT1 expression level was significantly reduced during complete remission. Long-term follow-up of WT1 expression level showed continuous or significant increase of WT1 expression in acute leukemia patients predicts refractoriness and impending relapse.Conclusion Quantification of WT1 expression level by RT-FQ-PCR may be used to predict relapse and monitor MRD for acute leukemia patients.