期刊
STEM CELLS
卷 33, 期 7, 页码 2114-2125出版社
WILEY
DOI: 10.1002/stem.2021
关键词
Cancer stem cell; Triple-negative breast cancer; NANOG; Junctional adhesion molecule-A; Fluorescent reporter system
资金
- NIH [R21 CA191263, GM50009, R25CA148052]
- Cleveland Clinic Research Program Committee
- Cleveland Clinic Foundation
- Case Comprehensive Cancer Center Pilot Grant
- Special Funds in Aging Cancer Energy Balance Research [P30 CA043703]
- American Cancer Society [IRG-91-022-15, IRG-91-022-18]
- Sam and Salma Gibara Fund
- Lerner Research Institute
- Case Comprehensive Cancer Center
- Sontag Foundation
- Voices Against Brain Cancer
- Blast GBM
- Ohio Cancer Research Associates
- NIH K99/R00 Pathway to Independence Award [CA157948, R01 (NS083629)]
- V Scholar Award from the V Foundation for Cancer Research
Advanced cancers display cellular heterogeneity driven by self-renewing, tumorigenic cancer stem cells (CSCs). The use of cell lines to model CSCs is challenging due to the difficulty of identifying and isolating cell populations that possess differences in self-renewal and tumor initiation. To overcome these barriers in triple-negative breast cancer (TNBC), we developed a CSC system using a green fluorescent protein (GFP) reporter for the promoter of the well-established pluripotency gene NANOG. NANOG-GFP+ cells gave rise to both GFP+ and GFP(-) cells, and GFP+ cells possessed increased levels of the embryonic stem cell transcription factors NANOG, SOX2, and OCT4 and elevated self-renewal and tumor initiation capacities. GFP+ cells also expressed mesenchymal markers and demonstrated increased invasion. Compared with the well-established CSC markers CD24(-)/CD44(+), CD49f, and aldehyde dehydrogenase (ALDH) activity, our NANOG-GFP reporter system demonstrated increased enrichment for CSCs. To explore the utility of this system as a screening platform, we performed a flow cytometry screen that confirmed increased CSC marker expression in the GFP+ population and identified new cell surface markers elevated in TNBC CSCs, including junctional adhesion molecule-A (JAM-A). JAM-A was highly expressed in GFP+ cells and patient-derived xenograft ALDH+ CSCs compared with the GFP(-) and ALDH(-) cells, respectively. Depletion of JAM-A compromised self-renewal, whereas JAM-A overexpression induced self-renewal in GFP(-) cells. Our data indicate that we have defined and developed a robust system to monitor differences between CSCs and non-CSCs in TNBC that can be used to identify CSC-specific targets for the development of future therapeutic strategies. Stem Cells. Stem Cells 2015;33:2114-2125
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