Abstract: Objective To explore the significance and interrelationship the protein expression of cyftoskeletal protein 4.1N, E-cadherin and β-catenin in the metastatic proliferation of gastric cancer. Methods We collected 55 cases of gastric carcinoma (adenocarcinoma) and 30 cases of gastric mucosa (distance from tumor edge ≥10 cm) as control study removed by surgical operation. The expressions of cytoskeletal protein 4.1N,E-cadherin and β-catenin protein were detected by Elivision immunohistochemicaly assay. Results The protein expression of cytoskeletal protein 4.1N, E-cadherin and β-catenin in gastric cancer group were significantly lower than that in control group (P<0.05), moreover, that in poorly-differentiated group was significantly lower than those in well- and moderately-differentiated groups (P<0.05). The protein expression of these three proteins between gastric cancer with and without lymph node metastasis group had significant difference (P<0.05). The IOD values of these three proteins were decreased with the increase of TNM stages in gastric cancer group. Similarly there was significant difference between the two groups (P<0.05). The expression of these proteins were not related with sex or age (P>0.05). There were positive correlation between cytoskeleton protein 4.1N and E-cadherin protein (r=0.381, P<0.05), E-cadherin protein and β-catenin (r=0.571, P<0.05), cytoskeleton protein 4.1N and β-catenin(r=0.602, P<0.05). Conclusion There were positive correlations among the expressions of cytoskeletal protein 4.1N, E-cadherin and β-catenin, indicating that important synergies exist among three proteins in promoting occurrence and development of gastric cancer.