Objective: To study the clinical pathologic characteristics of β-catenin, Ki67 and Her-2/neu in gastric cancer and the correlation of β-catenin and Ki67 to the protein expression and gene conditions of Her-2/Neu. Methods: The protein expression of β-catenin, Ki67 and Her-2/Neu was detected by immunohistochemistry in 101 cases of gastric cancer and the gene conditions of Her-2/Neu by fluorescence in situ hybridization (FISH). Results: The protein expression ofβ-catenin, Ki67 and Her-2/Neu had close relationship with the clinical pathologic characteristics of gastric cancer. The β-catenin and Ki67 had obvious correlation to the differentiation, infiltration and lymphatic metastasis of the gastric cancer (P<0.05). The Ki67 had close relationship with the tumor-node-metastasis staging staging of gastric cancer (P<0.05). Her-2/Neu had close relationship with the differentiation and tumor-node-metastasis staging of gastric cancer (P<0.05) but had no relationship with the infiltration and lymphatic metastasis of the gastric cancer (P<0.05). The protein expression of Ki67 had significantly positive correlation to the protein expression and gene amplification conditions of Her-2/Neu (r=0.567, P<0.05 for protein; r=0.304, P<0.05 for gene). Conclusions: Combined detection of β-catenin, Ki67 and Her-2/Neu can be used as a reliable method to help the observation of biological behavior, diagnosis and prognosis of gastric cancer, and Ki67 can be used to serve the preliminary screening of Her-2/Neu gene state.