Koletsis Nikolaos E, Mangani Sylvia, Franchi Marco, Piperigkou Zoi, Karamanos Nikos K
Biochemistry, Biochemical Analysis & Matrix Pathobiology Research Group, Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece.
Department for Life Quality Studies, University of Bologna, 47921 Rimini, Italy.
Cells. 2025 Aug 30;14(17):1351. doi: 10.3390/cells14171351.
Breast cancer (BC) remains a leading cause of cancer-related mortality in women. Extracellular matrix (ECM) remodeling is a critical modulator of tumor invasion and metastasis. Three-dimensional (3D) cell culture models have been proposed as advanced systems better mimicking the tumor microenvironment (TME), potentially offering enhanced insights into underlying mechanisms compared to conventional two-dimensional (2D) cultures. This study highlights how BC cells develop metastatic potential and tumor progression independently from ECM contact using advanced 3D spheroid culture models compared to traditional 2D cultures in triple-negative breast cancer (TNBC) cell lines. Spheroids were formed using ultra-low adhesion plates, and their morphological and functional properties were assessed via phase-contrast and scanning electron microscopy (SEM), along with functional assays. Both cell lines formed compact spheroids exhibiting mesenchymal-to-epithelial transition (MET) characteristics. Functional assays showed enhanced cell migration and dissemination of spheroid-derived cancer cells. Gene expression profiling revealed increased expression of ECM remodeling enzymes, cell surface receptors, and adhesion molecules in 3D cultures compared to 2D. MicroRNA analysis highlighted distinct regulatory patterns specifically associated with metastasis and epithelial-to-mesenchymal transition (EMT). These findings demonstrate that 3D spheroid models effectively recapitulate the complexity of TNBC, providing valuable insights into ECM dynamics, epigenetic regulation, and metastatic behavior and potentially guiding improved therapeutic strategies.
乳腺癌（BC）仍然是女性癌症相关死亡的主要原因。细胞外基质（ECM）重塑是肿瘤侵袭和转移的关键调节因子。三维（3D）细胞培养模型已被提出作为更能模拟肿瘤微环境（TME）的先进系统，与传统的二维（2D）培养相比，可能能更深入地洞察潜在机制。本研究强调了与三阴性乳腺癌（TNBC）细胞系中的传统2D培养相比，使用先进的3D球体培养模型，BC细胞如何独立于ECM接触而产生转移潜能和肿瘤进展。使用超低吸附板形成球体，并通过相差显微镜和扫描电子显微镜（SEM）以及功能测定来评估其形态和功能特性。两种细胞系均形成了表现出间充质向上皮转化（MET）特征的紧密球体。功能测定显示球体来源的癌细胞的细胞迁移和扩散增强。基因表达谱分析显示，与2D培养相比，3D培养中ECM重塑酶、细胞表面受体和粘附分子的表达增加。微小RNA分析突出了与转移和上皮向间充质转化（EMT）特别相关的独特调控模式。这些发现表明，3D球体模型有效地概括了TNBC的复杂性，为ECM动态、表观遗传调控和转移行为提供了有价值的见解，并可能指导改进治疗策略。
