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Background: Gastric cancer is one of the most common cancers in the world. Programmed death-ligand 1 (PD-L1) is the main ligand of the programmed death-1 receptor that plays a key role in apoptosis in patients with gastric cancer. Materials and Methods: In this study, forty gastric cancer patients and forty healthy controls were enrolled. Serum PD-L1 was measured by enzyme-linked immunosorbent assay. Results: The mean (interquartile range) value of expression level of PD-L1 was 70.6 (36.6–127.3) in patients and 47.7 (29.9–92.1) in controls. There were no significant differences in PD-L1 expression between the two groups and tumor characteristics, but there was a statistically significant difference between the high expression level of PD-L1 and gastric cancer development (P = 0.044, odds ratio = 2.77, 95% confidence interval = 1.12–6.86). Conclusions: PD-L1 level increases in gastric cancer patients, so it could be used as a predictor factor.