The Suvar, Enhancer of zeste, and Trithorax (SET), myeloid-Nervy-DEAF-1 (MYND) domain-containing protein 3 (SMYD3), was reported to be upregulated in various tumors. However, its role in pancreatic cancer progression remains unclear.To explore the role of SMYD3 in the pancreatic cancer.The expressions of SMYD3, caspase-3, and matrix metallopeptidase-2 (MMP-2) were detected in pancreatic cancer and non-tumor tissues by immunohistochemistry. The CCK-8 and transwell assays were performed to test proliferation, migration, and invasion ability in short hairpin RNA (shRNA-SMYD3) pancreatic cancer cell line SW1190. RT-PCR and Western blot were used to detect the expressions of SMYD3, caspase-3, and MMP-2 in SW1990 cell line and shRNA-SMYD3 SW1990 cell line.The expressions of SMYD3, caspase-3, and MMP-2 were upregulated in pancreatic cancer. The SMYD3 was positively associated with caspase-3 and MMP-2 expressions in pancreatic cancer tissues. SMYD3, TNM stages, histological differentiation, and lymph node metastasis were identified as an independent prognostic factor. Moreover, interfered SMYD3 expression in SW1990 cell line significantly reduced the cell proliferation, migration, and invasion. RT-PCR and Western blot showed the expression of MMP-2 decreased in shRNA-SMYD3 SW1990 cell line, but no significant change was observed in the caspase-3 expression.The overexpression of SMYD3 promoted proliferation, migration, and invasion of pancreatic cancer, and SMYD3 may affect the pancreatic cancer progression by regulating MMP-2 rather than caspase-3.