Development of Quantitative RT-PCR for the Expression of Wilms’ Tumor WT1 Suppressor Gene in Leukemia on the LightCycler

Abstract

Wilms’ tumor (WT) is a childhood neoplasm which arises as a result of inactivation of the WT1 gene. The WT1 suppressor gene encodes a transcription factor of the zinc finger family. Recently, Miwa et al. [1] reported that WT1 was expressed in human leukemia. In addition, Inoue et al. [2] reported that WT1 was a new prognostic factor and a new marker for the detection of minimal residual disease (MRD) in acute leukemia. However, their methods were qualitative, or at best, semi-quantitative. Since then, several quantitative RT-PCR methods using external standards and competitors have appeared but are labor intensive and timecomsuming. We have developed a new technique to quantify WTI expression in bone marrow cells in acute leukemia and myelodysplastic syndrome (MDS) using LightCycler technology.