Detecting phospho-signaling protein of bone marrow leukemia cells by phospho-signaling flow cytometry / 中国实验血液学杂志
Journal of Experimental Hematology
;
(6): 1176-1179, 2011.
Article
in Chinese
| WPRIM
| ID: wpr-261906
ABSTRACT
The purpose of this study was to establish the phospho-specific flow cytometry (phospho-flow) to detect the phosphorylated signaling proteins of leukemia cells and to evaluate its useful value in leukemia study. The bone marrow of leukemia children was collected, and treated by phospho-flow of extracted mononuclear cells (MNC) and phospho-flow of directly fixed bone marrow (BM) respectively. In phospho-flow of extracted MNC, the MNC extracted from BM were fixed and permeabilized, then were cultured with P-AKT and P-ERK1/2, finally were analyzed by flow cytometry. In phospho-flow of directly fixed BM, the BM was treated with fixation/lysis buffer and 90% methanol, then were incubated with the surface CD antibody, P-AKT and P-ERK1/2, finally the treated BM cells were analyzed by flow cytometry. The results showed that the positive rates of P-AKT and P-ERK1/2 in MNC treated by phospho-flow of extracted MNC of 26 leukemia children were 46.2% and 30.8% respectively, while the positive rates of P-AKT and P-ERK1/2 in BM treated by phospho-flow of directly fixed BM were 50.0% and 38.5% respectively. The comparison of positive rates of P-AKT and P-ERK1/2 between the 2 treatment protocol showed no difference (p > 0.05). It is concluded that the phospho-flow of directly fixed BM established by our laboratory can be used to analyze the signaling proteins of leukemia cells.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Bone Marrow
/
Bone Marrow Cells
/
Signal Transduction
/
Leukemia
/
MAP Kinase Signaling System
/
Cell Biology
/
Proto-Oncogene Proteins c-akt
/
Flow Cytometry
/
Metabolism
/
Methods
Type of study:
Practice guideline
Limits:
Child
/
Child, preschool
/
Female
/
Humans
/
Infant
/
Male
Language:
Chinese
Journal:
Journal of Experimental Hematology
Year:
2011
Type:
Article
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