Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences-WULS, Nowoursynowska 159, 02-776 Warsaw, Poland.
BMC Vet Res. 2012 Mar 27;8:35. doi: 10.1186/1746-6148-8-35.
It is supposed that fibroblasts present in tumour microenvironment increase cancer invasiveness and its ability to metastasize but the mechanisms have not been clearly defined yet. Thus, the current study was designed to assess changes in gene expression in five various cancer cell lines grown as a co-culture with the carcinoma-associated fibroblasts (CAFs) in vitro.
A carcinoma-associated fibroblast cell line was isolated from a canine mammary cancer. Then, a co-culture of cancer cells with the CAFs was established and maintained for 72 hrs. Having sorted the cells, a global gene expression in cancer cells using DNA microarrays was examined. The analysis revealed an up-regulation of 100 genes and a down-regulation of 106 genes in the cancer cells grown as a co-culture with the CAFs in comparison to control conditions. The PANTHER binomial statistics tool was applied to determine statistically over-manifested pathways (p < 0.05). Bulk of the up-regulated genes are involved in the adhesion, the angiogenesis, the epithelial-mesenchymal transition (EMT) and generally take part in the developmental processes. These results were further confirmed using real-time qPCR. Moreover, a wound-healing assay and growth characteristics on Matrigel matrix showed that CAFs increase cancer cell migration and matrix invasion.
The results of the current study showed that the co-culturing of cancer cells and the CAFs caused significant changes to the cancer gene expression. The presence of the CAFs in a microenvironment of cancer cells promotes adhesion, angiogenesis and EMT.
据推测，肿瘤微环境中的成纤维细胞会增加癌症的侵袭性及其转移能力，但目前尚未明确其机制。因此，本研究旨在评估五种不同的癌细胞系在体外与癌相关成纤维细胞（CAFs）共培养时基因表达的变化。
从犬乳腺肿瘤中分离出一种癌相关成纤维细胞系。然后，建立了癌细胞与 CAFs 的共培养物，并维持 72 小时。对细胞进行分选后，使用 DNA 微阵列检查了癌细胞的全基因表达。与对照条件相比，在与 CAFs 共培养的癌细胞中，有 100 个基因上调，106 个基因下调。应用 PANTHER 二项式统计工具确定了统计学上明显表现的途径（p < 0.05）。大量上调的基因参与细胞黏附、血管生成、上皮-间充质转化（EMT），并普遍参与发育过程。这些结果通过实时 qPCR 进一步得到证实。此外，划痕愈合实验和在 Matrigel 基质上的生长特性表明，CAFs 可促进癌细胞迁移和基质侵袭。
本研究结果表明，癌细胞与 CAFs 的共培养会导致癌细胞基因表达发生显著变化。CAFs 在癌细胞微环境中的存在促进了细胞黏附、血管生成和 EMT。
