Objective To investigate the expression of urokinase-type plasminogen activator (uPA) mRNA in gastric cancer tissues and cancer-adjacent tissues and the relationship between its expression and biologic behavior of tumor. Methods Fourty-eight cases with gastric cancer were detected for the expression of uPA mRNA by fluorogenic probe quantitative reverse transcription polymerase chain reaction (RTPCR). Results The positive expression rate of uPA mRNA was 83.3%, 25.0%, 93.8% and 62.5% in gastric cancer tissues,cancer-adjacent tissues, gastric cancer tissues with lymph node metastasis and with non-lymph node metastasis respectively. Expression of uPA mRNA was positively related with the invasion depth of gastric cancer. Conclusion Expression of uPA mRNA is significantly increased in gastric cancer and it can be used as an indicator to judge the metastasis and prognosis of tumor.