Expression patterns of WT-1 and Bcr-Abl measured by TaqMan quantitative real-time RT-PCR during follow-up of leukemia patients with the Ph chromosome

( views:74, downloads:0 )
Jaspal Kaeda()
Sue Saunders()
John M Goldman()
Journal Title:
Volume , Issue 07, 2004
Key Word:
WT-1;Bcr-Abl;real-time RT-PCR;chronic myelogenous leukemia

Abstract´╝Ü Background This study was designed to quantitatively measure WT-1 expression levels in patients with chronic myelogenous leukemia (CML) and acute lymphoblastic leukemia (ALL) during follow-up and to clarify the value of WT-1 as a molecular marker in minimal residual disease monitoring.Methods The TaqMan quantitative real-time RT-PCR method was established by using cloned WT-1 cDNA or synthesized oligonucleotides resembling WT-1 cDNA fragments in limit dilution as template until a stable and reliable standard curve was obtained. In a 25-month follow-up, the transcriptional levels of WT-1, Bcr-Abl, and Abl gene, were quantitatively measured in bone marrow cells from 25 CML or acute lymphoblastic leukemia (ALL) patients with the Ph chromosome. In addition, the expression of these genes in 40 samples of normal peripheral blood was also examined using the same method. The ratios of WT-1/Abl and Bcr-Abl/Abl were both plotted, and the two expression patterns were compared as well as their clinical significance.Results The levels of WT-1 expression in normal peripheral blood were detectable. In CML and Ph positive ALL patients, WT-1 expression levels changed in parallel with the Bcr-Abl expression pattern as the disease progressed or responded to effective treatment.Conclusion WT-1 expression provides a novel molecular marker in addition to Bcr-Abl for monitoring minimal residual disease (MRD) and targeting therapy in Ph chromosome-positive leukemia patients.

  • [1]Chen ZX. The possible role and application of WT-1 in human leukemia. Int J Hematol 2001;73:39-46.
  • [2]Brieger J, Weidmann E, Fenchel K, et al. The expression of the Wilm's tumor gene in acute myelocytic leukemias as a possible marker for leukemic blast cells. Leukemia 1994;8:2138-2143.
  • [3]Inoue K, Sugiyama H, Ogawa H, et al. WT-1 as a new prognostic factor and a new marker for the detection of minimal residual disease in acute leukemia. Blood 1994;84:3071-3979.
  • [4]Menssen HD, Renkl HJ, Rodeck U, et al. Presence of Wilms' tumor gene (wt1) transcripts and the WT-1 nuclear protein in the majority of human acute leukemias. Leukemia 1995;9:1060-1067.
  • [5]Bergmann L, Miething C, Maurer U, et al. High levels of Wilms' tumor gene (wt1) mRNA in acute myeloid leukemias are associated with a worse long-term outcome. Blood 1997;90:1217-1225.
  • [6]Schmid D, Heinze G, Linnerth B, et al. Prognostic significance of WT-1 gene expression at diagnosis in adult de novo acute myeloid leukemia. Leukemia 1997;11:639-643.
  • [7]Maurer U, Brieger J, Weidmann E, et al. The Wilms' tumor gene is expressed in a subset of CD34+ progenitors and downregulated early in the course of differentiation in vitro. Exp Hematol 1997;25:945-950.
  • [8]Gaiger A, Schmid D, Heinze G, et al. Detection of the WT1 transcript by RT-PCR in complete remission has no prognostic relevance in de novo acute myeloid leukemia. Leukemia 1998;12:1886-1894.
  • [9]Trka J, Kalinova M, Hrusak O, et al. Real-time quantitative PCR detection of WT1 gene expression in children with AML: prognostic significance, correlation with disease status and residual disease detection by flow cytometry. Leukemia 2002;16:1381-1389.
  • [10]Cilloni D, Gottardi E, De Micheli D, et al. Quantitative assessment of WT-1 expression by real-time quantitative PCR may be a useful tool for monitoring minimal residual disease in acute leukemia patients. Leukemia 2002;16:2115-2121.
  • [11]Cilloni D, Saglio G. Usefulness of quantitative assessment of WT-1 expression in CML patients undergoing Imatinib therapy. Semin in Hematol 2002;40(suppl):37-41.
WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615