Rama-Esendagli Dorina, Esendagli Gunes, Yilmaz Guldal, Guc Dicle
Department of Basic Oncology, Hacettepe University Cancer Institute, Sihhiye, 06100, Ankara, Turkey.
Mol Biol Rep. 2014 May;41(5):2885-92. doi: 10.1007/s11033-014-3144-3. Epub 2014 Jan 28.
Interactions with stromal components influence the growth, survival, spread, and colonization capacities of tumor cells. Fibroblasts and macrophages which are responsible for the stroma production and maintenance are of the basic elements found in tumor microenvironment. Cellular density and ratio of stromal cells to tumor cells can also have modulatory effects in cancer. Here, the contribution of fibroblast and/or macrophage cells on the malignant behavior of breast cancer cells was modeled in co-culture systems. Co-cultures were established at different cell densities and ratios with 4T1 breast cancer, NIH/3T3 or 3T3-L1 fibroblast, and J774A.1 monocyte/macrophage cell lines. Flow cytometry-based proliferation, 3D growth on alginate matrix, and matrigel invasion assays were performed to determine the change in the malignant assets of tumor cells. The data were also supported by immunocytochemical and morphological analyses. Co-culturing with fibroblasts (especially, NIH/3T3 cells) significantly supported the proliferation, scattering, and invasiveness of 4T1 cells whereas inclusion of macrophages disrupted this positive influence. On the other hand, the invasion capacity of 4T1 cells was not enhanced in the co-cultures with fibroblasts whose motility were inhibited with pertussis toxin pretreatment. Particularly at low-density seeding in 3D cultures, 4T1 cells could form substantially more spheroids than that of in the co-cultures with fibroblasts. Only, increasing the amount of fibroblasts could restore the 3D-growth. Intriguingly, co-existence of macrophage, fibroblast, and tumor cells in 3D cultures provided a convenient stroma sustaining the spheroid formation and growth. In conclusion, fibroblasts can form a favorable environment for tumor cells' spread and motility whereas restricting their 3D-growth capacity. On the other hand, presence of macrophages may disrupt the influence of fibroblasts and enhance the spheroid formation by the tumor cells.
与基质成分的相互作用会影响肿瘤细胞的生长、存活、扩散和定植能力。负责基质产生和维持的成纤维细胞和巨噬细胞是肿瘤微环境中的基本组成部分。基质细胞的细胞密度以及与肿瘤细胞的比例在癌症中也可能具有调节作用。在此，在共培养系统中模拟了成纤维细胞和/或巨噬细胞对乳腺癌细胞恶性行为的影响。以不同的细胞密度和比例，将4T1乳腺癌细胞、NIH/3T3或3T3-L1成纤维细胞以及J774A.1单核细胞/巨噬细胞系建立共培养体系。进行基于流式细胞术的增殖分析、在藻酸盐基质上的三维生长分析以及基质胶侵袭分析，以确定肿瘤细胞恶性特性的变化。免疫细胞化学和形态学分析也支持了这些数据。与成纤维细胞（尤其是NIH/3T3细胞）共培养显著促进了4T1细胞的增殖、分散和侵袭能力，而巨噬细胞的加入则破坏了这种积极影响。另一方面，在与经百日咳毒素预处理而运动能力受到抑制的成纤维细胞共培养时，4T1细胞的侵袭能力并未增强。特别是在三维培养中低密度接种时，4T1细胞形成的球体比与成纤维细胞共培养时要多得多。只有增加成纤维细胞的数量才能恢复三维生长。有趣的是，在三维培养中巨噬细胞、成纤维细胞和肿瘤细胞的共存提供了一个有利于球体形成和生长的基质。总之，成纤维细胞可以为肿瘤细胞的扩散和运动形成一个有利的环境，同时限制其三维生长能力。另一方面，巨噬细胞的存在可能会破坏成纤维细胞的影响，并增强肿瘤细胞的球体形成。
