Previous studies have shown that miRNAs participate in a wide range of biological functions and play important roles in various human diseases including cancer. We found miR-146b-5p significantly dysregulated in human pancreatic cancer cells by qRT-PCR. To demonstrate its function and regulation mechanism, we overexpressed miR-146-5p by transfecting the mimics. Our data showed that miR-146b-5p overexpression significantly reduced the abilities of migration and invasion of MIA PaCa-2 pancreatic cancer cells. Furthermore, we found that matrix metalloproteinase 16 (MMP16) was a downstream target of miR-146b-5p by dual-luciferase reporter assay. Altogether, our findings suggest that miR-146b-5p may be involved in pancreatic cancer cell migration and invasion by targeting MMP16, and miR-146b-5p may be a potential therapeutic target for the pancreatic cancer.