We assessed factors affecting fluorine F 18 fluorodeoxyglucose (F-FDG) uptake of metastatic lymph nodes (LNs) in advanced gastric cancer (AGC) due to low F-FDG uptake of metastatic LNs in gastric cancer. Retrospective analyses were performed on 31 patients with AGCs who underwent preoperative F-FDG positron emission tomography (PET) and subsequent gastrectomy. Metastatic LNs were compared with primary tumors (on a one-to-one basis) with respect to maximum standardized uptake values, glucose transporter type 1 (GLUT-1) expression, proliferation indices (using Ki-67), microvessel density, and lymphatic vessel density. Maximum standardized uptake values of metastatic LNs were significantly correlated with % GLUT-1 expression (ρ = 0.80, P < 0.0001) and Ki-67 labeling index (ρ = 0.57, P = 0.001) in LNs. These uptake values were also significantly correlated with SUVs (ρ = 0.54; P = 0.002), % GLUT-1 expression (ρ = 0.71, P < 0.0001), and Ki-67 labeling index (ρ = 0.43, P = 0.019) in primary tumors. In multiple regression analysis, only % GLUT-1 expression in primary tumors and metastatic LNs were significant factors in predicting maximum standardized uptake value of metastatic LNs. In AGCs, GLUT-1 expression and Ki-67 labeling index are important factors in predicting F-FDG uptake by metastatic LNs.