Pancreatic　cancer　(PC)　is　recognized　as　the　most　aggressive　and　deadliest　malignancy　because　it　has　the　highest　mortality　of　all　cancers　in　humans.　Mutations　in　multiple　tumor　suppressors　and　oncogenes　have　been　documented　to　be　involved　in　pancreatic　cancer　progression　and　metastasis.　The　upregulation　of　tetraspanin　1　(TSPAN1),　a　transmembrane　protein,　has　been　reportedly　observed　in　many　human　cancers.　However,　the　role　of　TSPAN1　and　its　underlying　molecular　mechanisms　in　PC　progression　have　not　been　fully　elucidated.　In　this　study,　we　validated　the　oncogenic　role　of　TSPAN1　in　PC,　showing　that　TSPAN1　reinforces　cell　proliferation,　migration,　invasion　and　tumorigenesis.　To　investigate　the　upregulation　of　TSPAN1　in　PC,　we　showed　that　miR-216a　is　the　upstream　negative　regulator　of　TSPAN1　via　direct　binding　to　the　TSPAN13＇-untranslated　region.　Through　RNA-Seq　analysis,　we　for　the　first　time　revealed　that　TSPAN1　expression　transcriptionally　regulates　ITGA2,　which　is　involved　in　the　actin　cytoskeleton　pathway.　The　stimulated　cell　proliferation　and　invasion　initiated　by　TSPAN　1　overexpression　could　be　abolished　by　knockdown　of　ITGA2　in　PC　cells.　Furthermore,　TSPAN1　epigenetically　regulates　the　expression　of　ITGA2　by　modulating　the　levels　of　TET2　DNMT3B　and　DNMT1,　resulting　in　hypomethylation　of　the　CpG　island　of　the　ITGA2　promoter.　In　conclusion,　the　newly　identified　miR-216a/TSPAN1/ITGA2　axis　is　involved　in　the　modulation　of　PC　progression　and　represents　a　novel　therapeutic　strategy　for　future　pancreatic　cancer　treatment.