荧光定量PCR检测急性白血病患者外周血 WT1基因表达及其临床意义

为研究WT1基因表达与临床疗效和预后的关系,探讨其在白血病微小残留病检测中的作用,用荧光定量RT-PCR方法检测55例初发白血病患者外周血及10例正常人外周血的WT1基因表达,跟踪20例急性白血病患者外周血WT1基因表达。结果显示,白血病初治组(40例ANLL、15例ALL)与正常人外周血WT1基因表达有显著差异(P<0.001)。在急性白血病患者中,WT1≤6.8×10-3组生存期长于WT1>6.8×10-3组(P=0.027);白血病患者初发时WT1呈高度表达,完全缓解后,迅速或缓慢下降至少1个对数级,复发时再次增高。跟踪检测20例急性白血病患者外周血WT1基因表达,结果7例复发,5例在临床复发前2-3月WT1的水平明显增高,至少上升0.8个对数级。结论:荧光定量RT-PCR方法检测白血病外周血WT1表达具有简便易行、准确性高、特异性好的特点。与正常人外周血比较,WT1在各类白血病外周血中呈高度表达,且表达水平与预后负相关;对外周血WT1基因表达进行定量分析,可用于微小残留病的监测。

Fluorescence Quantitative PCR Detection of WT1 Gene Expression in Peripheral Blood of Patients with Acute Leukemias and Its Clinical Implications

To elucidate the expression of WT1 in all types of leukemias and its implications for monitoring minimal residual disease in patients with acute leukemia, the peripheral blood from 55 leukemia patients and 10 normal voluteer was detected by using FQ-RT-PCR. Follow-up monitoring of WT1 expression of peripheral blood was performed for 20 patients with acute leukemia. The results showed that the expression of WT1 gene in all types of leukemias was significantly higher than that in normal control (P < 0.001). For ANLL and ALL patients, the survival time in the group of WT1 6.8 x 10(-3), (P = 0.027). Follow-up detection of the expression of WT1 in peripheral blood samples from 20 acute leukemia patients, 7 cases relapsed after complete remission has been done. In 5 of 7 relapsed patients, the expression of WT1 had obviously increased about 2 - 3 months before clinical relapse became apparent. It is concluded that the established FQ-RT-PCR method is accurate and specific. The expression of WT1 gene is relatively high in all types of leukemias compared with normal peripheral blood cells, the higher WT1 expression may associate with poor prognosis in acute leukemia, and the dynamics of WT1 level correlate with the disease status. The quantitative assessment of WT1 expression in peripheral blood samples by FQ-RT-PCR may be a useful tool for monitoring minimal residual disease.