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Logo of bmcvetresBioMed Centralsearchsubmit a manuscriptregisterthis articleBMC Veterinary Research
 
BMC Vet Res. 2012; 8: 16.
Published online Feb 21, 2012. doi:  10.1186/1746-6148-8-16
PMCID: PMC3315417
Global gene expression profiles of canine macrophages and canine mammary cancer cells grown as a co-culture in vitro
Magdalena Król,corresponding author1 Karol M Pawłowski,1,2 Kinga Majchrzak,1 Małgorzata Gajewska,1 Alicja Majewska,1 and Tomasz Motyl1
1Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences - WULS, Nowoursynowska 159, 02-776 Warsaw, Poland
2Department of Animal Environment Biology, Faculty of Animal Sciences, Warsaw University of Life Sciences - WULS, Ciszewskiego 8, 02-786 Warsaw, Poland
corresponding authorCorresponding author.
Magdalena Król: magdalena_krol/at/sggw.pl; Karol M Pawłowski: karol_pawlowski/at/sggw.pl; Kinga Majchrzak: kinga.majchrzak24/at/wp.pl; Małgorzata Gajewska: malgorzata_gajewska/at/sggw.pl; Alicja Majewska: alamaj/at/poczta.onet.pl; Tomasz Motyl: tomasz_motyl/at/sggw.pl
Received July 28, 2011; Accepted February 21, 2012.
Abstract
Background
Solid tumours comprise various cells, including cancer cells, resident stromal cells, migratory haemopoietic cells and other. These cells regulate tumour growth and metastasis. Macrophages constitute probably the most important element of all interactions within the tumour microenvironment. However, the molecular mechanism, that guides tumour environment, still remains unknown. Exploring the underlying molecular mechanisms that orchestrate these phenomena has been the aim of our study.
A co-culture of canine mammary cancer cells and macrophages was established and maintained for 72 hrs. Having sorted the cells, gene expression in cancer cells and macrophages, using DNA microarrays, was examined. The results were confirmed using real-time qPCR and confocal microscopy. Moreover, their ability for migration and invasion has been assessed.
Results
Microarray analysis showed that the up-regulated genes in the cancer cell lines are involved in 15 highly over-manifested pathways. The pathways that drew our diligent attention included: the inflammation pathway mediated by chemokine and cytokine, the Toll receptor signalling pathway and the B cell activation. The up-regulated genes in the macrophages were involved in only 18 significantly over-manifested pathways: the angiogenesis, the p53 pathway feedback loops2 and the Wnt signalling pathway. The microarray analysis revealed that co-culturing of cancer cells with macrophages initiated the myeloid-specific antigen expression in cancer cells, as well as cytokine/chemokine genes expression. This finding was confirmed at mRNA and protein level. Moreover, we showed that macrophages increase cancer migration and invasion.
Conclusions
The presence of macrophages in the cancer environment induces acquisition of the macrophage phenotype (specific antigens and chemokines/cytokines expression) in cancer cells. We presumed that cancer cells also acquire other myeloid features, such as: capabilities of cell rolling, spreading, migration and matrix invasion (what has also been confirmed by our results). It may, perhaps, be the result of myeloid-cancer cell hybrid formation, or cancer cells mimicking macrophages phenotype, owing to various proteins secreted by macrophages.
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