Endometrial Cancer Spheres Show Cancer Stem Cells Phenotype and Preference for Oxidative Metabolism
This study aimed to characterize endometrial cancer regarding cancer stem cells (CSC) markers, regulatory and differentiation pathways, tumorigenicity and glucose metabolism. Endometrial cancer cell line ECC1 was submitted to sphere forming protocols. The first spheres generation (ES1) was cultured in adherent conditions (G1). This procedure was repeated and was obtained generations of spheres (ES1, ES2 and ES3) and spheres-derived cells in adherent conditions (G1, G2 and G3). Populations were characterized regarding CD133, CD24, CD44, aldehyde dehydrogenase (ALDH), hormonal receptors, HER2, P53 and β-catenin, fluorine-18 fluorodeoxyglucose ([18F]FDG) uptake and metabolism by NMR spectroscopy. An heterotopic model evaluated differential tumor growth. The spheres self-renewal was higher in ES3. The putative CSC markers CD133, CD44 and ALDH expression were higher in spheres. The expression of estrogen receptor (ER)α and P53 decreased in spheres, ERβ and progesterone receptor had no significant changes and β-catenin showed a tendency to increase. There was a higher 18F-FDG uptake in spheres, which also showed a lower lactate production and an oxidative cytosol status. The tumorigenesis in vivo showed an earlier growth of tumours derived from ES3. Endometrial spheres presented self-renewal and differentiation capacity, expressed CSC markers and an undifferentiated phenotype, showing preference for oxidative metabolism.
KeywordsEndometrial neoplasms Neoplastic stem cells Glucose metabolism
American Type Culture Collection
basic fibroblast growth factor
bovine serum albumin solution
counts per minute
cancer stem cells
human endometrioid carcinoma type I cell line
epidermal growth factor
epithelial to mesenchymal transition
first sphere generation
second sphere generation
third sphere generation
first generation of adherent cells derived from the spheres
second generation of adherent cells derived from the spheres
third generation of adherent cells derived from the spheres
hematoxylin and eosin
mean fluorescence intensity
Rooswell Park Memorial Institute 1640 Medium
Tris-buffered saline Tween-20
uniformly enriched 13C isotopomer glucose
This study was funded by the Foundation for Science and Technology, Portugal, through individual support to Carvalho MJ (SFRH/SINTD/60068/2009), by the Portuguese Society of Gynecology through the 2016 Research Prize and by CIMAGO. CNC.IBILI is supported through the Foundation for Science and Technology, Portugal (UID/NEU/04539/2013), and co-funded by FEDER-COMPETE (POCI-01-0145-FEDER-007440).
The NMR spectrometer is part of the National NMR Network and was purchased as part of the Portuguese National Programme for Scientific Re-equipment (REDE/1517/RMN/2005), with funds from POCI 2010 (European Fund for Regional Development) and from the Foundation for Science and Technology, Portugal. The authors thank to the Pathology Service of the University Hospital Centre of Coimbra for technical support and David Anthony Tucker for the manuscript review.
Compliance with Ethical Standards
Conflits of Interest
Nothing to declare.
The experimental protocol was approved by the Ethics Committee of the Medicine Faculty of Coimbra University (Ref: Of IBB/48/09). All experiments were performed in accordance with guidelines and regulations of the European Union.
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