地高辛掺入RTPCR半定量检测WT1基因在AML中的表达及其意义  

Analysis of WT1 gene expression in acute myelocytic leukemia by DIG labeled RT PCR.

作　　者：王玲[1] 王玮[2] 傅建新[2] 陈子兴[2] 

机构地区：[1]青岛医学院附属第二医院,266042 [2]苏州医学院附属第一医院江苏省血液研究所,215006

出　　处：《江苏医药》2000年第10期781-783,共3页Jiangsu Medical Journal

基　　金：国家自然科学基金!(39770 306);江苏省卫生厅科研基金!(H9703)

摘　　要：目的　建立客观的半定量检测WT1基因表达的方法 ,探讨WT1基因表达水平与急性髓细胞性白血病 (AML)预后的关系。方法　用地高辛标记的脱氧三磷酸尿苷 (DIG 11 dUTP)掺入逆转录聚合酶链反应 (RT PCR)的方法检测 ,用KG 1细胞建立WT1表达的半定量稀释曲线 ,分析5 3例AML及 2 1例非恶性疾患病人骨髓中WT1基因的表达。结果　倍比稀释的KG 1细胞WT1基因表达水平与稀释倍数有良好的线性关系 ;将WT1基因与 β actin基因的表达水平之比 <0 16定为 (- ) ,≥ 0 16～ <0 6为 (+ ) ,≥ 0 6～ <1 0为 ( ) ,≥ 1 0为 ( )。 2 1例非恶性疾患病人骨髓中WT1基因均为阴性 ,41例初治AML病人WT1基因表达阳性率为 92 7% ,随着WT1基因表达水平的升高 ,完全缓解 (CR)率有明显下降的趋势 (<0 6及≥ 0 6组比较P <0 0 2 5 )。CR后WT1基因表达水平明显下降 ,复发后又显著升高。结论　DIG掺入RT PCR半定量检测WT1基因表达的方法客观、准确 。Objective To establish an objective and quantitative criterion for the assessment of WT1 gene expression and study the correlation between WT1 gene expression and prognosis of AML.Methods Using DIG labeled RT PCR assay,a semi quantitative curve of WT1 gene expression was established in KG 1 cell line.Based on this standard,WT1 gene expression was determined in the bone marrow specimens from 53 AML patients and 21 nonmalignant patients.Results WT1 gene expression level was classified as (-)(the ratio of WT1/β actin PCR products<0 16),(+)(≥0 16～<0 6),()(≥0 6～<1 0)and ()(≥1 0) respectively.WT1 gene expression in 21 nonmalignant patients was (-),positive rate of WT1 gene expression was 92 7% in 41 first diagnosed cases.The CR rate was markedly decreased as WT1 gene expression level increased(≥0 6 v <0 6 group, P< 0 025).Short term follow up of 12 M3 patients showed that WT1 gene expression level was markedly decreased after CR whereas highly increased when relapsed.Conclusion The method we established using DIG labeled RT PCR was objective and quantitative.WT1 gene expression level was correlated with the prognosis of AML.

关 键 词：急性髓细胞性白血病 地高辛 半定量RT-PCR WT1基 

分 类 号：R733.71[医药卫生—肿瘤]