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Open Access Highly Accessed Research article

The gene expression profiles of canine mammary cancer cells grown with carcinoma-associated fibroblasts (CAFs) as a co-culture in vitro

Magdalena Król1*, Karol M Pawłowski12, Katarzyna Szyszko1, Henryk Maciejewski3, Izabella Dolka4, Elisabetta Manuali5, Michał Jank1 and Tomasz Motyl1

Author affiliations

1 Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences - WULS, Nowoursynowska 159, 02-776 Warsaw, Poland

2 Department of Animal Environment Biology, Faculty of Animal Sciences, Warsaw University of Life Sciences - WULS, Ciszewskiego 8, 02-786 Warsaw, Poland

3 Institute of Computer Engineering, Control and Robotics I-6, Wroclaw University of Technology, Wybrzeże Wyspiańskiego 27, 50-320 Wroclaw, Poland

4 Department of Pathology and Veterinary Diagnostics, Faculty of Veterinary Medicine, Warsaw University of Life Sciences - WULS, Nowoursynowska 159, 02-776 Warsaw, Poland

5 Area Diagnostica Integrata Istologia e Microscopia Elettronica Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche, Via G. Salvemini 1, 06126 Perugia, Italy

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Citation and License

BMC Veterinary Research 2012, 8:35  doi:10.1186/1746-6148-8-35

Published: 27 March 2012

Abstract

Background

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.

Results

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.

Conclusion

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.