Santos Ana Rita M P, Kirkpatrick Bruce E, Kim Mirim, Anseth Kristi S, Park Yongdoo
Department of Biomedical Science, College of Medicine, Korea University, Seoul 02841, Republic of Korea.
Department of Chemical and Biological Engineering and the BioFrontiers Institute, University of Colorado, Boulder, Colorado 80303. USA.
Acta Biomater. 2024 Dec;190:264-272. doi: 10.1016/j.actbio.2024.10.031. Epub 2024 Oct 28.
The tumor microenvironment (TME) comprises diverse cell types within an altered extracellular matrix (ECM) and plays a pivotal role in metastasis through intricate cell-cell and cell-ECM interactions. Fibroblasts, as key constituents of the TME, contribute significantly to cancer metastasis through their involvement in matrix deposition and remodeling mechanisms, modulated by their quiescent or activated states. Despite their recognized importance, the precise role of fibroblasts in cancer cell invasion remains incompletely understood. In this study, we investigated the impact of fibroblast activity on cancer cell progression using a 2D co-culture model. Michigan Cancer Foundation-7 (MCF7) breast cancer cells were co-cultured with normal human lung fibroblasts (NHLF), both with and without transforming growth factor β (TGFβ) treatment. Traction force microscopy (TFM) was employed to quantify traction and velocity forces associated with cellular migration. We observed that TGFβ-activated fibroblasts form a distinctive ring around cancer cells in co-culture, with increased traction and tension at the cell island boundary. This force distribution is associated with the localization of force-related proteins at these boundary regions, including vinculin and E-cadherin. Metabolic profiling revealed a strong OXPHOS signal specific to the activated fibroblasts, in contrast to normal fibroblasts, which primarily display migratory behavior and a more heterogeneous pattern of forces and metabolic activity in co-culture. Our findings offer valuable insights into the mechanical forces and metabolic dynamics governing cellular migration in the tumor microenvironment, where our co-culture model could complement in vivo studies and enable researchers to explore specific microenvironmental cues for a deeper understanding of TME mechanisms. STATEMENT OF SIGNIFICANCE: Cancer models mimicking the dynamics of tumor microenvironment (TME) are an ideal tool to study cancer mechanisms and treatment. However, the full understanding of how cancer cells interact with their surroundings and other cells is still unknown. To tackle this, we developed a simple yet effective 2D co-culture model that allows us to control the arrangement of cell cultures precisely and use various imaging techniques to study interactions between cancer cells and fibroblasts. Here we could measure cell movements, force distribution, metabolic activity, and protein localization and interplay those factors in vitro. Our model helps us observe the underlying mechanisms between cancer cells and fibroblasts, contributing to our understanding of the dynamics in the TME.
肿瘤微环境（TME）由细胞外基质（ECM）改变时的多种细胞类型组成，通过复杂的细胞间和细胞与ECM的相互作用在转移中起关键作用。成纤维细胞作为TME的关键组成部分，通过参与由其静止或激活状态调节的基质沉积和重塑机制，对癌症转移有显著贡献。尽管它们的重要性已得到认可，但成纤维细胞在癌细胞侵袭中的确切作用仍未完全了解。在本研究中，我们使用二维共培养模型研究了成纤维细胞活性对癌细胞进展 的影响。密歇根癌症基金会-7（MCF7）乳腺癌细胞与正常人肺成纤维细胞（NHLF）进行共培养，分别在有和没有转化生长因子β（TGFβ）处理的情况下。采用牵引力显微镜（TFM）来量化与细胞迁移相关的牵引力和速度力。我们观察到，在共培养中，TGFβ激活的成纤维细胞在癌细胞周围形成一个独特的环，在细胞岛边界处牵引力和张力增加。这种力的分布与这些边界区域力相关蛋白的定位有关，包括纽蛋白和E-钙黏蛋白。代谢谱分析显示，与正常成纤维细胞相比，激活的成纤维细胞有强烈的氧化磷酸化信号，正常成纤维细胞在共培养中主要表现出迁移行为以及更异质的力和代谢活性模式。我们的研究结果为肿瘤微环境中控制细胞迁移的机械力和代谢动力学提供了有价值的见解，我们的共培养模型可以补充体内研究，并使研究人员能够探索特定的微环境线索，以更深入地了解TME机制。重要性声明：模拟肿瘤微环境（TME）动态的癌症模型是研究癌症机制和治疗的理想工具。然而，对癌细胞如何与其周围环境和其他细胞相互作用的全面理解仍然未知。为了解决这个问题，我们开发了一个简单而有效的二维共培养模型，使我们能够精确控制细胞培养的排列，并使用各种成像技术研究癌细胞与成纤维细胞之间的相互作用。在这里，我们可以测量细胞运动、力的分布、代谢活性和蛋白质定位，并在体外将这些因素相互关联。我们的模型帮助我们观察癌细胞与成纤维细胞之间的潜在机制，有助于我们理解TME中的动态变化。
