Gastric cancer (GC) is a malignant disease with high incidence and mortality rates worldwide. Nephronectin (NPNT) was found to be dysregulated in some kinds of cancer. The goal of our study was to explore the expression profile of NPNT based on large numbers of GC samples with detailed clinicopathological and prognostic data from our institution and the data from a public database. A total of 117 GC samples and 73 corresponding non-tumorous adjacent tissues (NATs) were obtained from GC patients and used to detect expression of NPNT through immunohistochemistry. Western blot and qRT-PCR were performed to examine expression of NPNT in GC cell lines. Our results found that the positive expression ratio of NPNT in GC tissues is significantly higher than that in NATs (p<0.001). Chi-squared analysis results showed positive expression ratio of NPNT was significantly associated with depth of tumor invasion (p=0.049) and TNM stage (p=0.017). Kaplan-Meier survival and cox analysis results showed that patients with positive NPNT protein expression tend to have poorer prognosis than those with negative NPNT expression (p=0.0032) and NPNT expression was independent prognostic factor. High expression level was seen in GC cell lines. Furthermore, through a series of cancer cell proliferation, invasion and migration associated experiments, we found that NPNT could evidently promote GC cell proliferation, invasion and migration, as well as epithelial-mesenthymal transition. In summary, NPNT was evidently overexpressed in GC and had an oncogenic role. In the future, NPNT could serve as a promising therapeutic target for treating GC patients.