The MAGEA gene family that encodes cancer testis antigens is differentially expressed in many cancers. Though MAGEA3 expression has been detected in gastrointestinal malignancies, its role in pancreatic ductal adenocarcinoma (PDAC) has not been well established. We assessed 57 patients who underwent intent-to-cure surgery for PDAC. Total RNA from paraffin-embedded pancreatic tumors was extracted and assessed forMAGEA3gene expression by an optimized probe-based quantitative real-time RT-PCR (qRT) assay.MAGEA3gene expression was detected by qRT in 25 (44%) patients. For the entire cohort, detection ofMAGEA3expression was associated with significantly decreased overall survival (median, 16vs33 months; log-rank,p= 0.032). When clinicopathologic factors, including age, gender, stage, tumor extent, lymph node metastasis, tumor grade, perineural invasion and lymphovascular invasion were assessed by univariate analysis,MAGEA3gene expression and tumor grade were significant prognostic factors for poor survival (HR 2.1, 95% CI: 1.0–4.4,p= 0.041; and HR 3.7, 95% CI: 1.8–7.6,p= 0.0004, respectively). Immunohistochemistry (IHC) was performed and confirmed MAGEA3 protein in PDAC specimens. In conclusion,MAGEA3is differentially expressed in patients with PDAC; its expression correlates with significantly worse survival. Molecular assessment for MAGEA3 should be considered to improve prognostic evaluation and to identify eligible patients for potential immune-based therapy.