实时定量RT-PCR检测急性白血病患者骨髓细胞WT1基因表达  

Detection of WT1 expression in bone marrow of acute leukemia patients with real time quantitative RT PCR

作　　者：顾伟英[1,2] 陈子兴[1] 曹祥山[2] 胡绍燕[1] 朱江[2] 王志林[2] 严峰[2] 王玮[1] 岑建农[1] 沈慧玲[1] 钱军[1] 

机构地区：[1]苏州大学附属第一医院,江苏省血液研究所 [2]苏州大学附属第三医院,常州市第一人民医院

出　　处：《中华血液学杂志》2004年第12期728-731,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金资助项目 (3 9770 3 0 6)

摘　　要：目的探讨急性白血病 (AL)患者骨髓细胞WT1基因表达水平。方法建立实时定量RT PCR方法 ,检测了 10 8例AL患者和 2 3例非白血病患者骨髓细胞WT1基因及内参照 β 胆色原脱氢酶 (GAPDH)基因的表达水平。结果初诊与复发AL患者骨髓细胞WT1基因中位表达水平经GAPDH校正后分别为 75 .10和 89.5 6 ,明显高于完全缓解组和对照组 (分别为 2 .0 7和 1.5 1) ,对照组与完全缓解组之间及初诊与复发组之间无统计学差异 ;70例初诊AL中急性淋巴细胞白血病、M1、M2 、M3 亚型WT1基因表达水平明显高于M5,即粒系表达明显高于单核系 ,且WT1基因表达水平与白细胞计数、骨髓原始细胞比例、mdr1基因表达无相关性 ,但与染色体核型有关。对其中 2例患者动态检测WT1基因表达可提示白血病复发情况。结论WT1基因在AL患者骨髓细胞高表达 ,可作为白血病疗效评价及监测残留病灶的指标。ObjectiveTo investigate Wilms’ tumor gene (WT1)expression levels in bone marrow(BM) of acute leukemia patients(ALs). MethodsA real time quantitative reverse transcriptase polymerase chain reaction (RQ RT PCR) method was established for detecting WT1 and internal reference GAPDH expression levels in BM of 108 ALs and 23 non leukemia controls by LightCycler. ResultsThe median expression levels of WT1 in 70 newly diagnosed ALs and 11 relapsed ALs were statistically higher than those in 23 ALs in complete remission(CR) and 23 non leukemic controls(75.10 and 89.56 vs 2.07 and 1.51 respectively). No statistic differences was found between the CR group and control group, nor between the newly diagnosed group and relapsed group. Of the 70 newly diagnosed ALs, median WT1 expression level of acute granulocytic leukemias was significantly higher than that of acute monocytic leukemias (M 5 ),but there was no statistic differences among the M 1 , M 2 , M 3 and ALL subtypes. Furthermore the WT1 levels were not correlated to peripheral WBC counts, BM blast percentage and multidrug resistant gene(mdr1) expression at presentation, but correlated to chromosome karyotypes.Dynamic analysis of WT1 levels of 2 patients on treatment showed that WT1 expression levels predicted relapse. Conclusion WT1 expression levels in ALs were strikingly higher than that in non leukemias. WT1 can be a marker for detecting MRD and evaluating therapy efficacy in leukemias.

关 键 词：WT1基因 骨髓细胞 患者 表达水平 AL 初诊 急性白血病 实时定量 GAPDH 高表达 

分 类 号：R733.7[医药卫生—肿瘤]