European Biophysics Journal

, Volume 48, Issue 7, pp 673–684 | Cite as

Phenotypic, structural, and ultrastructural analysis of triple-negative breast cancer cell lines and breast cancer stem cell subpopulation

  • Milene Pereira Moreira
  • Fábio André Brayner
  • Luiz Carlos Alves
  • Geovanni Dantas Cassali
  • Luciana Maria SilvaEmail author
Original Article


Triple negative breast cancer (TNBC) is a highly heterogeneous disease, which influences the therapeutic response and makes difficult the discovery of effective targets. This heterogeneity is attributed to the presence of breast cancer stem cells (BCSCs), which determines resistance to chemotherapy and subsequently disease recurrence and metastasis. In this context, this work aimed to evaluate the morphological and phenotypic cellular heterogeneity of two TNBC cell lines cultured in monolayer and tumorsphere (TS) models by fluorescence and electron microscopy and flow cytometry. The BT-549 and Hs 578T analyses demonstrated large phenotypic and morphological heterogeneity between these cell lines, as well as between the cell subpopulations that compose them. BT-549 and Hs 578T are heterogeneous considering the cell surface marker CD44 and CD24 expression, characterizing BCSC mesenchymal-like cells (CD44+/CD24), epithelial cells (CD44/CD24+), hybrid cells with mesenchymal and epithelial features (CD44+/CD24+), and CD44/CD24 cells. BCSC epithelial-like cells (ALDH+) were found in BT-549, BT-549 TS, and Hs 578T TS; however, only BT-549 TS showed a high ALDH activity. Ultrastructural characterization showed the heterogeneity within and among BT-549 and Hs 578T in monolayer and TS models being formed by more than one cellular type. Further, the mesenchymal characteristic of these cells is demonstrated by E-cadherin absence and filopodia. It is well known that tumor cell heterogeneity can influence survival, therapy responses, and the rate of tumor growth. Thus, molecular understanding of this heterogeneity is essential for the identification of potential therapeutic options and vulnerabilities of oncological patients.


Cancer stem cell Cell line Cellular heterogeneity Triple negative breast cancer 



This work was financially supported through grants from Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) [grant number CBB-APQ-00054–12], Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© European Biophysical Societies' Association 2019

Authors and Affiliations

  1. 1.Serviço de Biologia Celular, Diretoria de Pesquisa e DesenvolvimentoFundação Ezequiel DiasBelo HorizonteBrazil
  2. 2.Laboratório de Imunopatologia Keizo Asami (LIKA)Universidade Federal de Pernambuco (UFPE)RecifeBrazil
  3. 3.Laboratório de Patologia Comparada, Departamento de Patologia Geral, Instituto de Ciências BiológicasUniversidade Federal de Minas GeraisBelo HorizonteBrazil
  4. 4.Instituto Aggeu Magalhães (IAM), Fundação Oswaldo Cruz (FIOCRUZ)RecifeBrazil

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