Objective To analyze the value of 64-detector-row CT combined with neuron-specific enolase (NSE) and progastrin-releasing peptide (ProGRP) in the differential diagnosis and TNM staging of lung cancer.Methods A total of 130 patients with highly suspected lung cancer who were present in the Department of Imaging of the Chuzhou First People's Hospital from May 2017 to October 2021 were included. All the patients underwent 64-detector-row spiral CT and the serum levels of NSE and ProGRP were detected. The histopathological diagnosis was set as the gold standard. The CT perfusion parameters and serum levels of NSE and ProGRP were compared between patients with lung cancer and those with benign pulmonary diseases. The receiver operating characteristic (ROC) curve was applied to analyze the diagnostic efficacy of CT perfusion parameters and serum levels of NSE and ProGRP for lung cancer. The CT perfusion parameters and serum levels of NSE and ProGRP were compared among lung cancer patients with different TNM stages. The diagnostic coincidence rate of TNM stages of lung cancer via CT perfusion parameters, serum levels of NSE and ProGRP and their combination was analyzed.Results There was no difference in BF between the lung cancer group and the benign pulmonary diseases group (P >0.05), while BV, PS and MTT were higher in the lung cancer group than those in the benign pulmonary diseases group (P < 0.05). The preoperative serum levels of NSE and ProGRP were higher in the lung cancer group than in the benign pulmonary diseases group (P < 0.05). The ROC curve analysis showed that the areas under the ROC curves (AUCs) of BV, PS, MTT, NSE, ProGRP and their combination for diagnosing lung cancer were 0.865 (95% CI: 0.775, 0.955), 0.861 (95% CI: 0.766, 0.956), 0.770 (95% CI: 0.602, 0.937), 0.875 (95% CI: 0.737, 0.962), 0.901 (95% CI: 0.843, 0.982), and 0.958 (95% CI: 0.916, 0.999), respectively. There was no difference in BF among patients with different TNM stages (P >0.05), while BV, PS and MTT were higher in patients with stage Ⅳ lung cancer than in others (P < 0.05). Besides, the serum levels of NSE and ProGRP were also higher in patients with stage Ⅳ lung cancer than in others (P < 0.05). The diagnostic coincidence rates of TNM stages of lung cancer via the combined detection were the highest, and were 91.67%, 88.89%, 100.00% and 100.00%, respectively for stage Ⅰ to Ⅳ.Conclusions The 64-detector-row spiral CT perfusion parameters combined with serum levels of NSE and ProGRP are effective in diagnosing lung cancer, and they improve the diagnostic coincidence rate of TNM stages of lung cancer.