摘要
目的以实时荧光定量PCR(FQ-PCR)检测WT1mRNA的方法,探讨WT1mRNA在白血病的治疗转归、判断预后及监测微小残留病灶(minimal residual disease,MRD)方面的作用.方法FQ-PCR扩增K562细胞的WT1基因和β2-MG,将扩增产物10倍梯度稀释制作标准曲线,定量检测64例白血病患者,56例对照者骨髓内WT1水平.结果(1)A组急性白血病(复发、初治、未缓解)及慢粒加速期和急变期病人,共40例,骨髓WT1表达较高,22.54～168.90拷贝/100 ngRNA;(2)B组缓解病人、慢粒慢性期,共24例,WT1 0～4.90拷贝/100 ngRNA;(3)C组非白血病患者和正常人骨髓,共56例,WT1表达较低,0～3.20拷贝/100 ngRNA.结论不同状态的白血病患者WT1表达有明显差异.WT1可作为疾病转归、疗效考核和MRD的指标.
Objective To investigate the role of WT1 gene in dectecting the treament result and the mininal residual disease （MRD） of acute leukemia （AL）. Methods Real -Time Fluorescence quantitative PCR was used to amplify β 2 -MG and WT1 gene from cultured K562 cells. Standard curves were constructed by performing Real-Time PCR on a standard template after 10-fold serial dilutions of cellular cDNA. Then we detected the WT1 mRNA levels of these samples from 64 leukemia patients and 56 controls. Results All the samples were divided to 3 groups. The first group of 40 patients with AL （relapsed, untreated and not remitted）, accelated phase or blast crisis of chronic granulocytic leukemia showed higher WT1 gene expression level, ranging from 22.54 - 165.90 copies/100 ng RNA. The second group of 24 patients （ remitted and chronic phase of chronic myeloid leukemia） showed WT1 expressions from 0 - 4.90 copise/100 ng RNA. The third group of 56 healthy people and other non-patients showed a much lower WT1 level, from 0 -3.20 copise/100 ng RNA. Conclusion The WT1 levels remain different in different disease phases. The gene can be used to detect the MRD, and correlate with the treatment results and prognosis of AL and CML
出处
《昆明医学院学报》
2008年第2期66-69,共4页
Journal of Kunming Medical College
