Endometrial Cancer Spheres Show Cancer Stem Cells Phenotype and Preference for Oxidative Metabolism.

Pathology oncology research : POR (2018-12-01)
Maria João Carvalho, Mafalda Laranjo, Ana Margarida Abrantes, João Casalta-Lopes, Daniela Sarmento-Santos, Tânia Costa, Beatriz Serambeque, Nuno Almeida, Telmo Gonçalves, Catarina Mamede, João Encarnação, Rui Oliveira, Artur Paiva, Rui de Carvalho, Filomena Botelho, Carlos Oliveira
RESUMO

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.

MATERIAIS
Número do produto
Marca
Descrição do produto

Sigma-Aldrich
Fetal Bovine Serum, non-USA origin, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Antibiotic Antimycotic Solution (100×), Stabilized, with 10,000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
hEGF, EGF, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
hBFGF, FGF-Basic, recombinant, expressed in E. coli, suitable for cell culture
Sigma-Aldrich
Poly(2-hydroxyethyl methacrylate), BioReagent, powder, suitable for cell culture
Sigma-Aldrich
Ponceau S, BioReagent, suitable for electrophoresis
Sigma-Aldrich
Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-74, ascites fluid
Sigma-Aldrich
Methyl cellulose, viscosity: 15 cP, BioReagent, suitable for cell culture
Sigma-Aldrich
Putrescine dihydrochloride, ≥98% (TLC)
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium/Nutrient Mixture F-12 Ham, With L-glutamine and 15 mM HEPES, without sodium bicarbonate, powder, suitable for cell culture