O-linked　N-acetylglucosamine　transferase　(OGT)　catalyzes　O-linked　glycosylation　(O-GlcNAcylation).　O-GlcNAcylation　is　a　post-translational　carbohydrate　modification　of　diverse　nuclear　and　cytosolic　proteins　by　the　addition　of　O-linked　beta-N-acetylglucosamine.　It　was　recently　demonstrated　that　OGT　and　the　level　of　O-GlcNAcylation　are　upregulated　in　esophageal　cancer;　however,　the　physiological　consequences　of　this　upregulation　remain　unknown.　The　current　study　reports　that　OGT　knockdown　by　short　hairpin　RNA　(shRNA)　did　not　affect　cell　viability;　however,　cell　migration　in　esophageal　cancer　Eca-109　cells　was　significantly　reduced.　OGT-specific　shRNA　vectors　efficiently　decreased　the　protein　and　mRNA　levels　of　OGT　and　the　RL2　level　(a　marker　of　O-GlcNAcylation　levels)　in　Eca-109　esophageal　cancer　cells.　In　addition,　colony　formation　and　cell　proliferation　assays　demonstrated　that　OGT-specific　shRNA　decreased　the　proliferation　of　Eca-109　cells;　however,　there　was　no　significant　statistical　difference　between　OGT-specific　shRNA　and　control　shRNA.　Notably,　transwell　assays　demonstrated　that　the　migratory　ability　of　Eca-109　cells　was　significantly　suppressed　following　knockdown　of　the　OGT　gene.　Correspondingly,　western　blot　analyses　demonstrated　that　OGT　knockdown　significantly　downregulated　the　expression　of　matrix　metalloproteinase　9　(MMP9)　in　Eca-109　cells.　These　results　suggest　that　OGT　may　promote　the　migration,　invasion　and　metastasis　of　esophageal　cancer　cells　by　enhancing　the　stability　or　expression　of　MMP9.