To investigate the localization of HBXIP protein over-expression in gastric adenocarcinoma, and its prognostic significance. HBXIP localization was detected by immunofluorescence in AGS gastric cancer cell line, and by immunohistochemical staining in 97 gastric adenocarcinomas, 41 adjacent non-tumor tissues and 13 gastric adenoma tissues. Correlation between HBXIP expression and clinicopathological features of gastric cancer patients was evaluated by Chi-square and Fisher's exact tests. Overall survival rates were calculated using Kaplan-Meier method. HBXIP was mainly expressed in the cytoplasm of gastric cancer. The positive and strongly positive expression rates of HBXIP protein in gastric cancers were 68.0% (66/97) and 49.5% (48/97) respectively, and were significantly higher than those in adjacent non-tumor tissues(48.8%, 20/41; 36.6%, 15/41) or gastric adenomas(2/13, 1/13; all <0.05). HBXIP expression correlated significantly with tumor differentiation and lymph node status (=0.007; 0.041). Kaplan-Meier survival analysis showed that the overall survival rate was significantly lower in gastric cancer patients with high HBXIP expression (=0.015). HBXIP expression in gastric cancer is mainly expressed in cytoplasmic, and the expression level is closely related to the prognosis. HBXIP expression status may potentially be used as an important prognostic indicator for gastric cancer.