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- Title
A flow cytometric method to detect apoptosis-related protein expression in minimal residual disease in acute myeloid leukemia<sup>1</sup>.
- Authors
van Stijn, A; Kok, A; van der Pol, M A; Feller, N; Roemen, G M J M; Westra, A H; Ossenkoppele, G J; Schuurhuis, G J
- Abstract
Minimal residual disease (MRD) cells are thought to be responsible for the persistence and relapse of acute myeloid leukemia (AML). Flow cytometric MRD detection by the establishment of a leukemia-associated phenotype (LAP) at diagnosis can be used in 80% of AML patients, allowing detection and functional characterization of MRD in follow-up bone marrow. One of the mechanisms contributing to inefficient chemotherapy is apoptosis resistance. Measuring apoptosis parameters in MRD cells will help to unravel the importance of apoptosis resistance in AML. We therefore developed a fourcolor flow cytometry method that enables establishment of apoptosis-related protein expression such as Bcl-2, Bcl-x[sub L], Mcl1 and Bax at diagnosis and in MRD. Firstly, validation of this assay using Western blot analysis in five leukemia cell lines showed a significant correlation (R=0.70: P<0.0001). Secondly, the influence of the permeabilization procedure on LAP expression was investigated in 38 AML samples at diagnosis and in 42 MRD samples. Quantification of the frequency of LAP+ cells with and without permeabilization showed no significant differences (diagnosis: P=0.57, follow-up: P=0.43). The flow cytometric protocol thus enables analysis of apoptosis-related proteins at different stages of the disease, which will lead to a better understanding of the role of apoptosis resistance in the emergence of MRD in AML.
- Subjects
ACUTE myeloid leukemia; PHENOTYPES; FLOW cytometry
- Publication
Leukemia (08876924), 2003, Vol 17, Issue 4, p780
- ISSN
0887-6924
- Publication type
Article
- DOI
10.1038/sj.leu.2402885