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The aim of this study was to investigate the expression and clinical significance of miR-155 and miR-181 in gastric cancer. Methods? Sixty-eight patients with gastric cancer and 80 healthy volunteers were selected as subjects. Serum samples of patients and volunteers were collected to detect miR-155 and miR-181 expression levels in serum and tumor tissues. ResuIts? miR-155 and miR-181 serum levels were significantly higher in patients with gastric cancer than those in the control group (P < 0.05). miR-155 and miR-181 expression levels in gastric cancer tissues were 16.74 ± 4.29 and 12.17 ± 3.26, respectively and 3.42 ± 0.39) and 3.06 ± 0.69 in paracancerous tissues, respectively, and they were 1.22 ± 0.21 and 1.08 ± 0.35 in normal tissues, respectively. miR-155 and miR-181 expression levels in cancer tissues were significantly higher than adjacent tissues, and they were lowest in normal tissues (P < 0.05). High miR-155 and miR-181 expression had significant effects on differentiation, T stage, lymph node metastasis, and clinical stage (P < 0.05). Cox multivariate analysis showed that onset, T stage, clinical stage, differentiation degree, lymph node metastasis, miR-155, and miR-181 were independent risk factors for prognosis of patients with gastric cancer. ConcIusion? Serum miR-155 and miR-181 have a diagnostic value in gastric cancer, and they are strong signals of poor prognosis.