Isolation of CD44~+/CD24~(-/low) and side population cells from MDA-MB-453 cells and the analysis of their activation of Wnt and Notch pathway
- VernacularTitle:MDA-MB-453细胞中分离CD44~+/CD24~(-/low)和SP细胞及其Wnt和Notch通路状态分析
- Author:
Xin LI
;
Jun REN
- Publication Type:Journal Article
- Keywords:
Breast neoplasms;
Receptor,epidermal growth factor;
Pluripotent stem cells;
Gene amplification;
Receptor,Notch1
- From:
Journal of Peking University(Health Sciences)
2003;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore stem cells phenotype of human epidermal growth factor receptor 2(HER2)-overexpressing breast cancer cells,MDA-MB-453 by isolation of CD44+CD24-/low and side population(SP).Analyze the relationship between SP cells and the gene amplification levels of the key proteins,?-catenin and Notch-1 in Wnt and Notch signal pathways.And study the correlation of SP cells and the gene amplification of HER2 and human epidermal growth factor receptor 3(HER3).Methods:CD44+CD24-/low and SP phenotype were studied in MDA-MB-453(HER2-overexpressing cell line)and MCF7(HER2 negative cell line)respectively by flow cytometry.The gene amplification of ?-catenin and Notch-1 in Wnt and Notch pathways were also studied by reverse transcriptase polymerase chain reaction(RT-PCR).The gene amplification levels of the proteins mentioned,HER2 and HER3 were roughly compared in MDA-MB-453 cells and their SP by RT-PCR.Results:There was no CD44+CD24-/low subpopulation in MDA-MB-453 cells,However,SP could be observed in MDA-MB-453.The gene amplification levels of ?-catenin and Notch-1 in Wnt and Notch pathways were both observed in MDA-MB-453 and MCF7.Their gene amplification levels were more evident in SP of MDA-MB-453 than in MDA-MB-453 cells.However,the similar gene amplification levels of HER2 and HER3 were observed.Conclusion:SP can be enriched in breast cancer stem cells of MDA-MB-453.Wnt and Notch pathways are activated in MDA-MB-453,which may be related to maintenance and activity of breast cancer stem cells of MDA-MB-453.
