The aim of this study is to explore the expression and significance of urokinase-type plasminogen activator (u-PA) and vascular endothelial growth factor (VEGF) in gastric cancer, providing a novel insight for the diagnosis and treatment of gastric cancer. The gastric cancer specimens, which were excised from 87 patients and confirmed during July, 2012–July, 2014, were selected as observation group, and the normal tissue next to the tumor (more than 5 cm from the edge of the tumor) from 45 patients were randomly selected as control. u-PA and VEGF were detected by immunohistochemistry for the analysis of the correlation of u-PA and VEGF in two groups. The positive rates of u-PA and VEGF in gastric cancer tissue were 81.61 and 79.31 %, respectively, which were 6.67 and 8.89 % in the control group, respectively. The positive rates in the observation group were obviously higher than those in the control group, and the difference was statistically significant (P < 0.05). Among the 87 gastric cancer tissue samples from the observation group, the positive rates of u-PA and VEGF in the gastric cancer with poor differentiation, lymphatic metastasis, invasion up to serosal layer, and TNM stage III + IV were obviously higher than those in the gastric cancer with high differentiation, non-lymphatic metastasis, invasion not up to the serosal layer, and TNM stage I + II, and the difference was statistically significant (P < 0.05). Among the 87 gastric cancer tissue samples from the observation group, u-PA and VEGF were found to be positive in 60 cases and negative in 7 cases. By comparing the two groups, u-PA and VEGF were positively correlated in gastric cancer tissue (P < 0.05). u-PA and VEGF were highly expressed in gastric cancer tissue, which could be used as the molecular biological indicators to predict the invasion and metastasis potential of gastric cancer. The combination of two factors plays a guiding role in early diagnosis and treatment of gastric cancer.