Kerst Gunter, Bergold Nina, Gieseke Friederike, Coustan-Smith Elaine, Lang Peter, Kalinova Marketa, Handgretinger Rupert, Trka Jan, Müller Ingo
Department of Pediatric Cardiology, University Children's Hospital, Tuebingen, Germany.
Am J Hematol. 2008 May;83(5):382-6. doi: 10.1002/ajh.21123.
In patients with acute leukemia, Wilms' tumor gene 1 (WT1) has been used as a target for the detection of minimal residual disease (MRD) by PCR techniques. The expression of WT1 protein, however, has not been extensively studied. To determine the relation between expression of WT1 transcripts and of the encoded protein, we examined leukemic cell lines and primary childhood leukemia samples using both real-time quantitative PCR (RQ-PCR) and flow cytometry. WT1 protein was highly expressed in the leukemic cell lines K562, HL-60, PLB 985, KG-1a and CEM. By contrast, 40 primary samples of acute lymphoblastic leukemia (ALL; B-ALL, n = 15 and T-ALL, n = 10) and acute myeloid leukemia (n = 15) expressed low levels of WT1 protein. RQ-PCR detected WT1 transcript levels in the same range as reported in earlier studies in childhood acute leukemia. The results of this study indicate the following: (i) there are considerable discrepancies between WT1 transcripts and protein expression; (ii) WT1 is not a suitable marker for flow cytometric MRD detection in childhood acute leukemia.
在急性白血病患者中,威尔姆斯瘤基因1(WT1)已被用作通过聚合酶链反应(PCR)技术检测微小残留病(MRD)的靶点。然而,WT1蛋白的表达尚未得到广泛研究。为了确定WT1转录本表达与编码蛋白表达之间的关系,我们使用实时定量PCR(RQ-PCR)和流式细胞术检测了白血病细胞系和儿童原发性白血病样本。WT1蛋白在白血病细胞系K562、HL-60、PLB 985、KG-1a和CEM中高表达。相比之下,40份急性淋巴细胞白血病(ALL;B-ALL,n = 15和T-ALL,n = 10)和急性髓细胞白血病(n = 15)的原发性样本表达低水平的WT1蛋白。RQ-PCR检测到的WT1转录本水平与早期儿童急性白血病研究中报道的范围相同。本研究结果表明:(i)WT1转录本与蛋白表达之间存在相当大的差异;(ii)WT1不是儿童急性白血病流式细胞术MRD检测的合适标志物。