Thymidylate synthase (TS) is a prognostic marker in various tumors. However, the results of previous investigations in gastric cancer (GC) were controversial. The objective of this article is to investigate whether TS expression is associated with clinical outcome in advanced GC receiving capecitabine alone chemotherapy. The study reviewed 58 cases of advanced GC in patients aged ≥65 years between December 2008 and June 2012. All patients were treated with capecitabine alone chemotherapy. Immunohistochemical staining for TS protein expression was performed. The relationships between TS expression and clinicopathological characteristics (included age, gender, number of metastatic sites, Eastern Cooperative Oncology Group (ECOG) score, differentiation, and lymph node metastatic status), chemotherapy response, progression-free survival (PFS), and overall survival (OS) were evaluated. There was no association between TS expression and age, gender, number of metastatic sites, ECOG score, differentiation, and lymph node metastatic status (P > 0.05). The chemotherapy response rates among patients with low- and high-level expression of TS protein were 52.0 % (13/25) and 21.2 % (7/33), respectively (χ (2) = 5.968, P = 0.015). The median PFS and OS in patients with low-level TS expression were significantly longer than those with high-level TS expression (PFS 8.0 vs 2.8 m, P = 0.001; OS 13.3 vs 7.9 m, P = 0.002, respectively). Multivariate Cox regression analysis revealed that TS expression was independent risk factor for OS (hazard ratio (HR) 0.237; 95 % confidence interval (CI) 0.108 to 0.520; P = 0.000). The present study demonstrates that TS expression is associated with chemotherapy response, PFS, and OS in advanced GC patients treated with capecitabine alone chemotherapy.