Extensive expression of craniofacial related homeobox genes in canine mammary sarcomas
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The global gene expression in three types of canine mammary tumors: carcinoma, fibrosarcoma and osteosarcoma were investigated by Affymetrix gene array technology. Unsupervised clustering analysis revealed a close clustering of the respective tumor types, with fibrosarcomas clustering close to the osteosarcomas and the carcinomas clustering closer to non-malignant mammary tissues (NMTs). A number of epithelial markers were expressed in both carcinomas and NMTs, whereas the sarcomas expressed genes related to mesenchymal differentiation. A comparison of the gene expression profile of the sarcomas versus carcinoma/NMTs revealed that the sarcomas, in particular the osteosarcomas, showed a striking upregulation of a panel of homeobox genes previously linked to craniofacial bone formation. In line with this finding, osteosarcomas showed an upregulation of bone morphogenetic proteins (BMPs) and of genes associated with retinoic acid signaling. Increased homeobox gene expression in sarcomas was also confirmed at the protein level by immunohistochemical analysis of tumor tissue, and in an osteosarcoma cell line after stimulation by BMP-2. These findings suggest that the development of mammary sarcomas specifically involves triggering of a set of homeobox genes related to neural crest and craniofacial bone development.
KeywordsCanine mammary tumors Mammary sarcoma Mammary osteosarcoma Homeobox transcription factor Gene expression profiling Craniofacial
Non-malignant mammary tissues
Bone morphogenetic protein
We would like to thank Maria Rydåker for technical assistance. This work was financed by a Strategic funding from the Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden to E. Hellmén.
H. Wensman, E. Hellmén, H. Göransson, A. Isaksson, G.R. Rutteman and N.E. Heldin designed the study. E. Hellmén did the pathological evaluation of the tumors. H. Wensman prepared the RNA and RNA quality controls. H. Göransson performed the array data normalization, background adjustments and statistical tests. H. Wensman set up the main direction for the manuscript and array data analysis and G. Pejler, H. Göranson, N.E. Heldin, E. Hellmén and A. Isaksson contributed to the data analysis. S. Strömberg and F. Pontén contributed with novel, validated antibodies. H. Wensman performed the IH verification and H. Wensman, E. Hellmén, S. Strömberg and F. Pontén evaluated the IH results and controls. K. Leuchowius performed the immunofluorescence and the evaluation of those results. H. Wensman and G. Pejler wrote the manuscript. All Authors approved the final version of the manuscript.
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