This study was aimed to investigate miR-216a expression in pancreatic cancer and determine its effects on proliferation. miR-216a was found downregulated in pancreatic cancer tissues as compared to benign pancreatic lesions. JAK2 was identified as a miR-216a gene target. Further, in vivo treatment of PANC-1 tumors with miR-216a reduced JAK2 protein levels in the tumor and reduced tumor volume. In conclusion, miR-216a may function as a tumor suppressor regulating pancreatic cancer cells by targeting the JAK/STAT pathway. Further studies with a larger number of patient samples are necessary to fully explore the diagnostic and therapeutic potential of miR-216a for pancreatic cancer.