Derlin-1, stanniocalcin-1, epithelial glycoprotein-2 (EGP-2) and maspin are associated with the metastasis of human breast cancer cells. This study reports the potential role of these molecules in metastasis of canine mammary tumours. Laser microdissected tissue samples were prepared from normal canine mammary gland and from simple adenomas, adenocarcinomas and their lymph node metastases. The expression of genes encoding the molecules of interest in these tissues was determined by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Most adenomas displayed reduced expression of derlin-1 mRNA. Few adenocarcinomas overexpressed derlin-1 mRNA, but all lymph node metastases overexpressed this gene product. Stanniocalcin-1 mRNA was not expressed within adenomas and was reduced in adenocarcinomas and their lymph node metastases. EGP-2 gene expression did not differ between normal, benign and malignant neoplastic tissues. Maspin gene expression varied markedly among the tumours with reduced or increased expression compared with normal mammary gland. Taken together, these results suggest that malignant behaviour of canine mammary adenocarcinoma is associated with reduced transcription of the stanniocalcin-1 gene and overexpression of the derlin-1 gene.