Introduction: The pancreatic ductal adenocarcinoma (PDAC) belongs to the most aggressive malignancies with an overall 5-year survival rate of less than 5%. A characteristic of this particular tumor entity is the early development of distant metastases, the mechanisms of which remain poorly understood. The aim of this study was to identify genes relevant to the process of metastasis in PDAC and analyze them for genetic and epigenetic alterations. Methods: We evaluated the metastatic and invasive potential of 16 PDAC cell lines of varying origin in a series of newly established orthotopic mouse models for PDAC. The resulting data was compiled into scores for metastatic and invasive potential. Methylation Specific PCR (MSP) and Bisulfite Sequencing PCR (BSP) were performed for all cell lines for the promoter region of 12 of the 15 described metastasis suppressor genes. The respective mRNA expression levels were determined by TaqMan Assay based qRT PCR for each gene and cell line as well as for a healthy control sample. Invasion- and metastasis-scores from the mouse models were compared to the data obtained from the epigenetic and mRNA analyses. Results: 5 of the 12 investigated metastasis suppressor genes showed promoter methylation in both BSP and MSP while the remaining ones did not show methylation. Results from both techniques were highly correlated (p<0.01-0.001) with correlation coefficients r>0.7 (except TIMP3). Promoter methylation correlated significantly with loss of mRNA expression in 2 of 5 methylated metastasis suppressor genes (p<0.05). Metastasis suppressor gene mRNA expression did not correspond with a less aggressive phenotype of PDAC: AKAP12, BRMS1, CD44, KAI1 and MASPIN were significantly overexpressed in PDAC cell lines compared to normal pancreatic tissue, whereas only GPR54, MAP2K4, TIMP3 and TXNIP showed expected reduced mRNA expression. Furthermore, a significant correlation between the mRNA expression levels and metastasis scores was detected for AKAP12 and MASPIN (p<0.05) and CD44 mRNA expression levels correlated significantly with invasion scores (p<0.05). Discussion: Metastasis suppressor genes have mostly been identified by analyzing their impact on a distinct and often single tumor entity, such as mammary carcinoma. Our data suggests that not all of these genes may play an according role in PDAC and that in fact some of them may even promote tumor invasion and metastasis. We suggest their function for PDAC be reinvestigated. Furthermore, epigenetic regulation by promoter hypermethylation appears not to be a key regulator for most metastasis suppressor genes in PDAC.