Background: Tumor markers CEA, CYFRA21-1, NSE, proGRP, and SCC-Ag were routinely used for lung cancer. The objective of this study was to evaluate the diagnostic and prognostic value of these markers.Methods: The levels of 5 serum tumor markers were measured in 255 patients with lung cancer (44 squamous cell carcinoma, 183 adenocarcinoma, 28 small cell lung cancer), 103 patients with benign lung disease, and 120 healthy controls. The relevant clinical data of the patients were collected to analyze the expression of CEA, CYFRA21-1, NSE, proGRP, and SCC-Ag in the serum of lung cancer and their correlation with prognosis.Results: The positive rates and expression levels of CEA, CYFRA21-1, NSE, proGRP, and SCC-Ag in the lung cancer group were higher than those in patients with benign lung diseases and healthy controls. CYFRA21-1 and SCC-Ag were highly expressed in lung squamous cell carcinoma, and NSE and proGRP were highly expressed in small cell lung cancer. The expression levels of CYFRA 21-1, NSE, and proGRP in small cell lung cancer were higher than those in non-small cell lung cancer. The levels of CEA, CYFARA21-1, NSE, and proGRP were linearly correlated with the occurrence of lung cancer, while the level of SCC-Ag was not significantly correlated with the occurrence of lung cancer. What’s more, the levels of CYFAR21-1 and NSE in the death group of lung cancer patients were higher than those in the survival group.Conclusion: CEA, CYFRA21-1, NSE, proGRP, and SCC-Ag played a good role in diagnosing lung cancer. CYFRA21-1 and SCC-Ag can be used as markers for the diagnosis of squamous cell carcinoma, and NSE and proGRP can be used as markers for small cell lung cancer. Meanwhile, the combined detection of CEA, CYFARA21-1, NSE, and proGRP had the highest diagnostic accuracy for lung cancer. Furthermore, CYFAR21-1 and NSE can be used to evaluate the prognosis of lung cancer patients.