Haartman Institute, POB 21, FI-00014 University of Helsinki, Finland.
Exp Cell Res. 2010 Jun 10;316(10):1739-47. doi: 10.1016/j.yexcr.2010.01.020. Epub 2010 Jan 22.
The role of paracrine tumor-stroma regulation in the progression of cancer is under intense investigation. Activated fibroblasts are key components of the tumor microenvironment providing the soluble factors mediating the regulation. Nemosis is an experimental model to study these parameters: formation of a multicellular spheroid activates fibroblasts and leads to increased production of soluble factors involved in the promotion of growth and motility. Role of nemosis was investigated in the tumorigenesis of HaCaT derivatives representing skin carcinoma progression. Conditioned medium from fibroblast spheroids increased proliferation rate of HaCaT derivatives. Expression of proliferation marker Ki-67 increased significantly in benign A5 and low-grade malignant II-4 cells, but did not further increase in the metastatic RT3 cells. Expression of p63, keratinocyte stem cell marker linked to cancer progression, was augmented by medium from nemotic fibroblasts; this increase was also seen in RT3 cells. Scratch-wound healing of the keratinocytes was enhanced in response to fibroblast nemosis. Neutralizing antibodies against growth factors inhibited wound healing to some extent; the response varied between benign and malignant keratinocytes. Migration and invasion were enhanced by conditioned medium from nemotic fibroblasts in benign and low-grade malignant cells. RT3 keratinocyte migration was further augmented, but invasion was not, indicating their intrinsic capacity to invade. Our data demonstrate that fibroblast nemosis increases proliferation and motility of HaCaT keratinocyte derivatives, and thus nemosis can be used as a model to study the role of soluble factors secreted by fibroblasts in tumor progression.
旁分泌肿瘤-基质调节在癌症进展中的作用正在受到深入研究。激活的成纤维细胞是肿瘤微环境的关键组成部分，提供了介导调节的可溶性因子。细胞融合是研究这些参数的实验模型：形成多细胞球体激活成纤维细胞，并导致参与促进生长和运动的可溶性因子的产生增加。细胞融合在代表皮肤癌进展的 HaCaT 衍生物的肿瘤发生中被研究。来自成纤维细胞球体的条件培养基增加了 HaCaT 衍生物的增殖率。增殖标志物 Ki-67 的表达在良性 A5 和低度恶性 II-4 细胞中显著增加，但在转移性 RT3 细胞中没有进一步增加。与癌症进展相关的角蛋白干细胞标志物 p63 的表达被融合成纤维细胞的培养基增强；在 RT3 细胞中也观察到这种增加。角蛋白细胞对成纤维细胞融合的划痕愈合反应增强。针对生长因子的中和抗体在一定程度上抑制了伤口愈合；良性和恶性角蛋白细胞之间的反应不同。来自融合成纤维细胞的条件培养基增强了良性和低度恶性细胞的迁移和侵袭。RT3 角蛋白细胞的迁移进一步增强，但侵袭没有增强，表明它们具有内在的侵袭能力。我们的数据表明，成纤维细胞融合增加了 HaCaT 角蛋白细胞衍生物的增殖和运动性，因此融合可以用作研究成纤维细胞分泌的可溶性因子在肿瘤进展中的作用的模型。
