A flow cytometric method to detect apoptosis-related protein expression in minimal residual disease in acute myeloid leukemia*
A flow cytometric method to detect apoptosis-related protein expression in minimal residual disease in acute myeloid leukemia*
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流式细胞术检测急性髓系白血病微小残留病中凋亡相关蛋白表达*
DOI：
10.1038/sj.leu.2402885
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发表时间：
2003
期刊：
影响因子：
11.4
通讯作者：
G. Schuurhuis
中科院分区：
文献类型：
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作者：
A. van Stijn;A. Kok;M. A. van der Pol;N. Feller;G M J M Roemen;A. Westra;G. J. Ossenkoppele;G. Schuurhuis
微小残留病（MRD）细胞被认为是导致急性髓性白血病（AML）持续存在和复发的原因。通过在诊断时建立白血病相关表型（AML）的流式细胞术MRD检测可用于80%的AML患者，允许在随访骨髓中检测MRD并进行功能表征。导致化疗无效的机制之一是凋亡抵抗。测量MRD细胞中的凋亡参数将有助于阐明AML中凋亡抵抗的重要性。因此，我们开发了一种四色流式细胞术方法，该方法能够在诊断和MRD时建立凋亡相关蛋白表达，如Bcl-2，Bcl-xL，Mcl-1和Bax。首先，在五种白血病细胞系中使用蛋白质印迹分析验证该测定法显示出显著相关性（R=0.70：P<0.0001）。其次，在诊断时的38个AML样品和42个MRD样品中研究了透化过程对BMP 1表达的影响。透化和未透化的PBMC+细胞频率的定量显示无显著差异（诊断：P= 0.57，随访：P= 0.43）。因此，流式细胞术方案能够在疾病的不同阶段分析凋亡相关蛋白，这将导致更好地理解细胞凋亡抵抗在AML中出现MRD中的作用。
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 four-color flow cytometry method that enables establishment of apoptosis-related protein expression such as Bcl-2, Bcl-xL, Mcl-1 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.
DOI：
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发表时间：
2000-05
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
I. Tamm;S. Kornblau;H. Segall;S. Krajewski;K. Welsh;S. Kitada;D. Scudiero;G. Tudor;Y. Qui;A. Monks;M. Andreeff;John Calvin Reed
通讯作者：
I. Tamm;S. Kornblau;H. Segall;S. Krajewski;K. Welsh;S. Kitada;D. Scudiero;G. Tudor;Y. Qui;A. Monks;M. Andreeff;John Calvin Reed
影响因子：
15.9
作者：
Milella, M;Kornblau, SM;Andreeff, M
通讯作者：
Andreeff, M