It has recently been found that the expression of the transglutaminase 3 (TGM3) is significantly reduced in esophageal cancer (EC). However, the potential of TGM3 as a useful biomarker or as a molecular target for EC diagnosis and treatment is unclear. The aim of the present study was to explore the role of the TGM3 in EC. The expression level of TGM3 were measured by real-time polymerase chain reaction and western blot analysis in EC cell lines, including SKGT-4, KYSE-510, OE33, OE21 and the normal esophageal epithelial cell line HEEC; fifty-eight pairs of tissues samples were also measured. through exogenous expression of TGM3 in EC cells it was found that the expression of TGM3 is closely associated with tumor cell growth and apoptosis. TGM3 was found downregulated in EC and closely associated with tumor proliferation and migration. In addition, overexpression of TGM3 apparently induces EC cell proliferation, migration, invasion and promotes cell apoptosis in vitro. Subsequent experiments identified the NF-κB signaling pathway as direct target of TGM3. Our data collectively demonstrate that TGM3 can be a candidate tumor suppressor that is able to induce EC cell proliferation and migration by downregulating the NF-κB signaling pathway, indicating that TGM3 may serve as a useful biomarker and therapeutic target for EC treatment.