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
Dans Chinois
| 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.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Moelle osseuse
/
Cellules de la moelle osseuse
/
Transduction du signal
/
Leucémies
/
Système de signalisation des MAP kinases
/
Biologie cellulaire
/
Protéines proto-oncogènes c-akt
/
Cytométrie en flux
/
Métabolisme
/
Méthodes
Type d'étude:
Guide de pratique
Limites du sujet:
Enfant
/
Enfant d'âge préscolaire
/
Femelle
/
Humains
/
Bébé
/
Mâle
langue:
Chinois
Texte intégral:
Journal of Experimental Hematology
Année:
2011
Type:
Article
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