Department of Public Health, University of Naples Federico II, 80131 Naples, Italy.
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, Italy.
Cells. 2019 Nov 14;8(11):1435. doi: 10.3390/cells8111435.
Melanoma is one of the most aggressive solid tumors and includes a stromal microenvironment that regulates cancer growth and progression. The components of stromal microenvironment such as fibroblasts, fibroblast aggregates and cancer-associated fibroblasts (CAFs) can differently influence the melanoma growth during its distinct stages. In this work, we have developed and studied a stromal microenvironment model, represented by fibroblasts, proto-myofibroblasts, myofibroblasts and aggregates of inactivated myofibroblasts, such as spheroids. In particular, we have generated proto-myofibroblasts from primary cutaneous myofibroblasts. The phenotype of proto-myofibroblasts is characterized by a dramatic reduction of α-smooth muscle actin (α-SMA) and cyclooxygenase-2 (COX-2) protein levels, as well as an enhancement of cell viability and migratory capability compared with myofibroblasts. Furthermore, proto-myofibroblasts display the mesenchymal marker vimentin and less developed stress fibers, with respect to myofibroblasts. The analysis of crosstalk between the stromal microenvironment and A375 or A2058 melanoma cells has shown that the conditioned medium of proto-myofibroblasts is cytotoxic, mainly for A2058 cells, and dramatically reduces the migratory capability of both cell lines compared with the melanoma-control conditioned medium. An array analysis of proto-myofibroblast and melanoma cell-conditioned media suggests that lower levels of some cytokines and growth factors in the conditioned medium of proto-myofibroblasts could be associated with their anti-tumor activity. Conversely, the conditioned media of melanoma cells do not influence the cell viability, outgrowth, and migration of proto-myofibroblasts from spheroids. Interestingly, the conditioned medium of proto-myofibroblasts does not alter the cell viability of both BJ-5ta fibroblast cells and myofibroblasts. Hence, proto-myofibroblasts could be useful in the study of new therapeutic strategies targeting melanoma.
黑色素瘤是最具侵袭性的实体肿瘤之一，包括调节癌症生长和进展的基质微环境。基质微环境的组成部分，如成纤维细胞、成纤维细胞聚集体和癌相关成纤维细胞（CAF），可以在黑色素瘤的不同阶段以不同的方式影响肿瘤的生长。在这项工作中，我们开发并研究了一种基质微环境模型，该模型由成纤维细胞、原肌成纤维细胞、肌成纤维细胞和失活肌成纤维细胞的聚集体（如球体）组成。特别是，我们从原代皮肤肌成纤维细胞中生成了原肌成纤维细胞。原肌成纤维细胞的表型特征是α-平滑肌肌动蛋白（α-SMA）和环氧化酶-2（COX-2）蛋白水平显著降低，与肌成纤维细胞相比，细胞活力和迁移能力增强。此外，与肌成纤维细胞相比，原肌成纤维细胞表达间充质标志物波形蛋白和发育不良的应力纤维。基质微环境与 A375 或 A2058 黑色素瘤细胞之间相互作用的分析表明，原肌成纤维细胞的条件培养基具有细胞毒性，主要针对 A2058 细胞，与黑色素瘤对照条件培养基相比，显著降低了两种细胞系的迁移能力。原肌成纤维细胞和黑色素瘤细胞条件培养基的基因表达谱分析表明，原肌成纤维细胞条件培养基中某些细胞因子和生长因子的水平较低可能与其抗肿瘤活性有关。相反，黑色素瘤细胞的条件培养基不会影响球体中原肌成纤维细胞的细胞活力、生长和迁移。有趣的是，原肌成纤维细胞的条件培养基不会改变 BJ-5ta 成纤维细胞和肌成纤维细胞的细胞活力。因此，原肌成纤维细胞可用于研究针对黑色素瘤的新治疗策略。
