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Mammospheres of hormonal receptor positive breast cancer diverge to triple-negative phenotype

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Mammospheres of hormonal receptor positive breast cancer diverge to triple-negative phenotype

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Wednesday, 10.01.2018

Authors and Affiliations:

Laranjo M1, Carvalho MJ2, Costa T3, Alves A3, Oliveira RC4, Casalta-Lopes J5, Cordeiro P3, Botas F3, Abrantes AM6, Paiva A7, Oliveira C8, Botelho MF6

1 Biophysics Institute, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; CIMAGO, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; CNC.IBILI, University of Coimbra, Coimbra, Portugal. 

2 Biophysics Institute, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; CIMAGO, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; Gynecology A Service, Coimbra Hospital and Universitary Centre, Coimbra, Portugal.

3 Biophysics Institute, Faculty of Medicine, University of Coimbra, Coimbra, Portugal.

4 Biophysics Institute, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; Pathology Service, Coimbra Hospital and Universitary Centre, Coimbra, Portugal.

5 Biophysics Institute, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; CIMAGO, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; Radiation Oncology Department, Coimbra Hospital and Universitary Centre, Coimbra, Portugal.

6 Biophysics Institute, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; CIMAGO, Faculty of Medicine, University of Coimbra, Coimbra, Portugal; CNC.IBILI, University of Coimbra, Coimbra, Portugal.

7 Cytometry Operational Management Unit, Clinical Pathology Service, Coimbra Hospital and Universitary Centre, Coimbra, Portugal.

8 CIMAGO, Faculty of Medicine, University of Coimbra, Coimbra, Portugal.

 

Abstract:

Objectives: This study aimed to characterize mammospheres from hormonal receptor (HR) positive and triple-negative breast cancer (TNBC), hypothesizing a differential profile of CSC and differentiation markers, and a stemness enrichment when successive sphere forming-protocols are performed.

Methods: Breast cancer cells MCF-7 and HCC1806 were submitted to sphere-forming protocols. The first sphere generation (MS1) was cultured in adherent conditions (G1). This procedure was repeated and generations of mammospheres (MS1, MS2, and MS3) and sphere-derived cells in adherent conditions (G1, G2, and G3) were obtained. The mammosphere forming capacity, self-renewal, area and doubling time were evaluated. Flow cytometry regarding CD133, CD24, and CD44 and western-blot regarding aldehyde dehydrogenase (ALDH), hormonal receptors and P53 expression was performed. Results: Breast cancer cell lines harboured the capacity to form spheres, which originated derived adherent populations. The sphere-forming capacity was enhanced in HCC1806-MS3 compared to MS1. Self-renewal was higher in MCF-7 mammospheres, which also had an increased area. The putative CSC markers CD133 showed tendency to be enhanced in mammospheres but the CD44þ/CD24-/low phenotype was not identified. The expression of ALDH was greater in mammospheres from MCF-7 and HCC1806 than in the respectively derived adherent cells. The expression of oestrogen receptor (ER)-a, progesterone receptor (PR) and P53 decreased in MCF-7 spheres. ER-b expression was lower in mammospheres from both cell lines compared with parental and derived adherent populations. Conclusions: Loss of HR and P53 expression in HR-positive mammospheres evidences the minor population of CSC which shares characteristics with the TNBC phenotype.

 

Journal: The Breast

 

Link: http://www.thebreastonline.com/article/S0960-9776(17)30779-8/abstract

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