Menssen H D, Renkl H J, Rodeck U, Maurer J, Notter M, Schwartz S, Reinhardt R, Thiel E
Department of Hematology and Oncology, Benjamin Franklin Klinik, Freie Universität Berlin, Germany.
Leukemia. 1995 Jun;9(6):1060-7.
The wt1 gene is located on chromosome 11p13 and encodes a zinc finger motif-containing transcription factor involved in regulation of growth and differentiation. Its expression was shown during embryonic development in various tissues as well as in a few human malignancies including acute leukemias. Using RT-PCR, we found wt1 gene expression in blast cells of the majority of 150 acute leukemia patients. Particularly, the wt1 transcript was detected in 12 of 14 (86%) pre-pre-B-ALL patients, in 33 of 41 (80%) cALL patients, in 23 of 31 (74%) T-ALL patients, and in 53 of 57 (93%) AML patients. Additionally, mononuclear cells from CML patients expressed the wt1 gene only when diagnosed with blast crisis. In contrast to acute human leukemias, mononuclear cells from reactive bone marrow (n = 4), and peripheral blood of healthy volunteers (n = 20), as well as normal peripheral CD34+ hematopoietic progenitors (n = 6) did not express the wt1 gene at detectable levels. Using the anti-WT1 MoAb 6F-H2 in an immunofluorescence assay on single cell level, we found the translated WT1 protein only in nuclei of leukemia blast cells but not in nuclei of normal CD34+ hematopoietic progenitor cells. Blast cells of 12 of 20 leukemia patients (60%) all tested positive for the wt1 gene expression by RT-PCR displayed a strong nuclear immunofluorescence. Its expression in the majority of human acute leukemias but not in normal mononuclear blood cells and normal CD34+ hematopoietic progenitors qualifies the wt1 gene transcript as a 'pan-acute leukemic' marker probably useful in monitoring minimal residual disease after chemotherapy and in detecting leukemic blast cells in purged or unpurged hematopoietic stem cell preparations intended to be used for autologous bone marrow transplantation.
wt1基因位于11号染色体p13区，编码一种含锌指基序的转录因子，参与生长和分化的调控。其表达在胚胎发育过程中的各种组织以及包括急性白血病在内的一些人类恶性肿瘤中均有显示。通过逆转录聚合酶链反应（RT-PCR），我们在150例急性白血病患者中的大多数原始细胞中发现了wt1基因表达。特别地，在14例前前B淋巴细胞白血病（pre-pre-B-ALL）患者中的12例（86%）、41例普通型急性淋巴细胞白血病（cALL）患者中的33例（80%）、31例T淋巴细胞白血病（T-ALL）患者中的23例（74%）以及57例急性髓系白血病（AML）患者中的53例（93%）中检测到了wt1转录本。此外，慢性粒细胞白血病（CML）患者的单核细胞仅在诊断为急变期时表达wt1基因。与人类急性白血病相反，反应性骨髓（n = 4）、健康志愿者外周血（n = 20）的单核细胞以及正常外周血CD34+造血祖细胞（n = 6）在可检测水平上均不表达wt1基因。在单细胞水平上使用抗WT1单克隆抗体6F-H2进行免疫荧光分析，我们发现翻译后的WT1蛋白仅存在于白血病原始细胞核中，而不存在于正常CD34+造血祖细胞核中。20例白血病患者中的12例（60%）通过RT-PCR检测wt1基因表达均呈阳性，其原始细胞显示出强烈的核免疫荧光。wt1基因在大多数人类急性白血病中表达，但在正常单核血细胞和正常CD34+造血祖细胞中不表达，这使得wt1基因转录本成为一种“泛急性白血病”标志物，可能有助于监测化疗后的微小残留病以及在检测用于自体骨髓移植的净化或未净化造血干细胞制剂中的白血病原始细胞。
