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Mammary Epithelial and Endothelial Cell Spheroids as a Potential Functional In vitro Model for Breast Cancer Research
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Development, Functional Characterization, and Matrix Effectors Dynamics in 3D Spheroids of Triple-Negative Breast

Nikolaos E Koletsis1, Sylvia Mangani1, Marco Franchi2

  • 1Biochemistry, Biochemical Analysis & Matrix Pathobiology Research Group, Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece.

Cells
|September 13, 2025
PubMed
Summary

Three-dimensional (3D) breast cancer models reveal metastatic potential independent of extracellular matrix contact. These advanced cultures offer new insights into tumor progression and therapeutic strategies.

Keywords:
3D breast cancer cell modelsbreast cancerepithelial-to-mesenchymal transitionestrogen receptor betaextracellular matrixmicroRNAsspheroids

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Area of Science:

  • Oncology
  • Cell Biology
  • Biotechnology

Background:

  • Breast cancer (BC) is a major cause of cancer mortality in women.
  • Extracellular matrix (ECM) remodeling influences tumor invasion and metastasis.
  • Three-dimensional (3D) cell cultures better mimic the tumor microenvironment (TME) than 2D cultures.

Purpose of the Study:

  • To investigate breast cancer cell metastatic potential and progression in 3D spheroid models compared to 2D cultures.
  • To analyze the role of ECM contact in tumor development using triple-negative breast cancer (TNBC) cell lines.
  • To identify molecular mechanisms underlying metastasis and tumor progression in 3D TNBC models.

Main Methods:

  • Triple-negative breast cancer (TNBC) cell lines were cultured in 3D spheroids using ultra-low adhesion plates.
  • Morphological and functional properties were assessed using phase-contrast microscopy, scanning electron microscopy (SEM), and functional assays.
  • Gene expression profiling and microRNA analysis were performed to compare 3D and 2D cultures.

Main Results:

  • 3D spheroids exhibited mesenchymal-to-epithelial transition (MET) characteristics.
  • Spheroid-derived cells showed enhanced migration and dissemination.
  • 3D cultures displayed increased expression of ECM remodeling enzymes, cell surface receptors, and adhesion molecules.
  • Distinct microRNA regulatory patterns associated with metastasis and epithelial-to-mesenchymal transition (EMT) were observed in 3D.

Conclusions:

  • 3D spheroid models effectively recapitulate TNBC complexity, including ECM dynamics and metastatic behavior.
  • These models provide valuable insights into epigenetic regulation and metastatic potential.
  • Findings may guide the development of improved therapeutic strategies for TNBC.