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A more accurate cut-off value of maximum standardized uptake value (SUVmax) in [18F]fluorodeoxyglucose positron emission tomography/computed tomography ([18F]FDG-PET/CT) is necessary to improve preoperative nodal staging in patients with lung cancer. Overall, 223 patients with lung cancer who had undergone [18F]FDG-PET/CT within 2 months before surgery were enrolled. The expression of glucose transporter-1, pyruvate kinase-M2, pyruvate dehydrogenase-E1α (PDH-E1α), and carbonic anhydrase-9 was evaluated by immunohistochemistry. Clinicopathological background was retrospectively investigated. According to PDH-E1α expression in primary lesion, a significant difference (p = 0.021) in SUVmax of metastatic lymph nodes (3.0 with PDH-positive vs 4.5 with PDH-negative) was found, but not of other enzymes. When the cut-off value of SUVmax was set to 2.5, the sensitivity and specificity were 0.529 and 0.562, respectively, and the positive and negative predictive values were 0.505 and 0.586, respectively. However, when the cut-off value of SUVmax was set according to PDH-E1α expression (2.7 with PDH-positive and 3.2 with PDH-negative), the sensitivity and specificity were 0.441 and 0.868, respectively, and the positive and negative predictive values were 0.738 and 0.648, respectively. The SUVmax cut-off value for metastatic lymph nodes depends on PDH-E1α expression in primary lung cancer. The new SUVmax cut-off value according to PDH-E1α expression showed higher specificity for [18F]FDG-PET in the diagnosis of lymph node metastasis. [ABSTRACT FROM AUTHOR]