Defined Extracellular Matrix Contexts Rewire 3D Breast Cancer Cell Models: Effects of Sulfated Hyaluronan in the Expression of Matrix Effector and Matrix-Based Cell Functional Properties
- 1Biochemistry, Biochemical Analysis & Matrix Pathobiology Research Group, Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece
Abstract
Background: Breast cancer progression is strongly influenced by tumor cell interactions with the extracellular matrix (ECM). Three-dimensional (3D) cell culture models provide more relevant systems for studying aspects of the tumor microenvironment (TME), including tumor architecture, matrix remodeling and cell-ECM communication. Collagen type I (Col I), fibronectin (FN) and hyaluronan (HA) are key ECM components involved in breast cancer progression. At the same time, integrins act as major matrix-sensing receptors that translate ECM cues into cellular responses. In this study, we investigated how defined ECM contexts regulate functional cell properties in shERβ MDA-MB-231 and Hs578T advanced 3D breast cancer cell models. Methods: Cells were cultured in 2D and 3D systems under matrix-free conditions or in Col I, FN, and mixed Col I/FN environments, in the presence or absence of HA or sulfated HA (sHA). Spheroid growth, matrix-dependent spheroid organization and dissemination were evaluated. In parallel, selected collagen- and fibronectin-associated integrin subunits, together with HA-related molecules including HAS2, CD44, and RHAMM, were analyzed to assess receptor-mediated matrix- and HA-associated responses. Results: Matrix substrates strongly influenced breast cancer spheroid behavior. Col I promoted outward dissemination, whereas FN-containing environments favored compact organization and restricted spreading. HA supported spheroid-like organization in a matrix-dependent manner, whereas sHA disrupted organized spheroid assembly in ECM-rich conditions, resulting in looser structures or smaller cell colonies, depending on the matrix context. Gene expression analysis revealed model-dependent remodeling of matrix- and HA-associated programs, with broader adhesion-related responses in shERβ MDA-MB-231 cells and stronger dimensionality-driven regulation in Hs578T cells. Conclusions: Overall, these findings demonstrate that defined ECM contexts actively regulate breast cancer spheroid behavior and matrix-associated receptor programs. Col I, FN and mixed Col I/FN environments generated distinct 3D phenotypes, affecting spheroid growth, organization and dissemination, while HA/sHA further modified these matrix-dependent responses. This study supports sHA as a functional matrix molecule with anti-cancer relevance and highlights the value of 3D matrix-based models for investigating tumor–matrix communication in breast cancer.
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BibTeX
@article{vetoulas2026defined,
title = {{Defined Extracellular Matrix Contexts Rewire 3D Breast Cancer Cell Models: Effects of Sulfated Hyaluronan in the Expression of Matrix Effector and Matrix-Based Cell Functional Properties}},
author = {Marios Vetoulas and Maria Drosinou and Nikoleta Alexiou and Angeliki Farmaki and Zoi Piperigkou and Nikos K. Karamanos},
journal = {Cancers},
year = {2026},
month = oct,
doi = {10.3390/cancers18203288},
url = {https://www.mdpi.com/2072-6694/18/20/3288},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Defined Extracellular Matrix Contexts Rewire 3D Breast Cancer Cell Models: Effects of Sulfated Hyaluronan in the Expression of Matrix Effector and Matrix-Based Cell Functional Properties
AU - Marios Vetoulas
AU - Maria Drosinou
AU - Nikoleta Alexiou
AU - Angeliki Farmaki
AU - Zoi Piperigkou
AU - Nikos K. Karamanos
PY - 2026
DA - 2026/10/12
JO - Cancers
DO - 10.3390/cancers18203288
UR - https://www.mdpi.com/2072-6694/18/20/3288
AB - Background: Breast cancer progression is strongly influenced by tumor cell interactions with the extracellular matrix (ECM). Three-dimensional (3D) cell culture models provide more relevant systems for studying aspects of the tumor microenvironment (TME), including tumor architecture, matrix remodeling and cell-ECM communication. Collagen type I (Col I), fibronectin (FN) and hyaluronan (HA) are key ECM components involved in breast cancer progression. At the same time, integrins act as major matrix-sensing receptors that translate ECM cues into cellular responses. In this study, we investigated how defined ECM contexts regulate functional cell properties in shERβ MDA-MB-231 and Hs578T advanced 3D breast cancer cell models. Methods: Cells were cultured in 2D and 3D systems under matrix-free conditions or in Col I, FN, and mixed Col I/FN environments, in the presence or absence of HA or sulfated HA (sHA). Spheroid growth, matrix-dependent spheroid organization and dissemination were evaluated. In parallel, selected collagen- and fibronectin-associated integrin subunits, together with HA-related molecules including HAS2, CD44, and RHAMM, were analyzed to assess receptor-mediated matrix- and HA-associated responses. Results: Matrix substrates strongly influenced breast cancer spheroid behavior. Col I promoted outward dissemination, whereas FN-containing environments favored compact organization and restricted spreading. HA supported spheroid-like organization in a matrix-dependent manner, whereas sHA disrupted organized spheroid assembly in ECM-rich conditions, resulting in looser structures or smaller cell colonies, depending on the matrix context. Gene expression analysis revealed model-dependent remodeling of matrix- and HA-associated programs, with broader adhesion-related responses in shERβ MDA-MB-231 cells and stronger dimensionality-driven regulation in Hs578T cells. Conclusions: Overall, these findings demonstrate that defined ECM contexts actively regulate breast cancer spheroid behavior and matrix-associated receptor programs. Col I, FN and mixed Col I/FN environments generated distinct 3D phenotypes, affecting spheroid growth, organization and dissemination, while HA/sHA further modified these matrix-dependent responses. This study supports sHA as a functional matrix molecule with anti-cancer relevance and highlights the value of 3D matrix-based models for investigating tumor–matrix communication in breast cancer.
SN - 2072-6694
ER -