荧光定量PCR检测WT_1基因在白血病中的表达

目的 :建立荧光定量 RT- PCR检测 WT1 基因的方法 ,探讨 WT1 基因在各类白血病中的表达。方法 :RT-PCR分别扩增 K5 6 2细胞 WT1 基因及β- actin基因 ,凝胶切割并回收、纯化 ,经紫外分光光度计定量后作为标准模板。采用荧光定量 RT- PCR方法检测 80例白血病患者骨髓或外周血及 10例正常人外周血中 WT1 基因表达。结果 :白血病患者组 (4 9例 AML、18例 AL L、7例 HAL、6例 CML )与正常人外周血 WT1 基因表达有显著差异 ,髓系表达高于淋巴系 ,M5显著低表达。结论 :荧光定量 RT- PCR方法检测 WT1 基因灵敏性高、特异性好。与正常人外周血比较 ,WT1 在各类型白血病中呈高度表达

Fluorescence quantitative PCR detection of WT1gene expression in leukemias

Objctive:To establish a fluorescense quantitative RT-PCR(FQ-RT-PCR)method for detection of WT 1 gene expression.To elucidate the expression of WT 1 in all types of leukemias.Methods:The conventional RT-PCR was used to amplify WT 1 gene and β-actin gene from K562 cells.After gel extraction,purification and quantification by a photometer,quantitative stand template was constructed.Using FQ-RT-PCR,we measured the peripheral blood or bone marrow samples from 80 leukemia patients and 10 normal voluteer′s peripheral blood samples.Results:The expression of WT 1 gene in all types of leukemias was significantly higher than that in normal controls.WT 1 expression levels was significantly higher in myeloid than in lymphoid.Patients with M 5 expressed significantly lower levels.Conclusion:The established FQ-RT-PCR method is sensitive and specific.The expression of WT 1 gene was relatively high in all types of leukemias compared with normal peripheral blood cells.