Objective To investigate the relationship of serum levels of carcinoembryonic antigen (CEA), carbohydrate antigen 199 (CA199), squamous cell carcinoma antigen (SCC-Ag), carbohydrate antigen 125 (CA125) and cytokeratin fragment antigen 21-1 (CYFRA21-1) with different pathological types and stages of non-small cell lung cancer(NSCLC) in elderly patients. Methods A total of 90 patients with age over 65 years and pathologically identified NSCLC hospitalized in our department from August 2009 to December 2012 were subjected in this study. Electrochemiluminescence method was applied to detect the expression of above-mentioned 5 serum tumor markers in these patients. Results The expression levels of the five serum tumor markers were significantly higher in the cohort than in healthy volunteers (P＜0.05). But no significant difference was found in the levels between different genders, or among those of different age groups (P＞0.05). The levels of CEA, CA199, and CA125 were significantly higher in patients with adenocarcinoma than those of squamous cell cancer (P＜0.05), but levels of SCC-Ag and CYFRA21-1 were significantly higher in patients with squamous cell cancer than in those of adenocarcinoma. (P＜0.05). The expression of CYFRA21-1 and CA125 was significantly higher in the patients at N2 and N3 stages than those at N0 and N1 stages (P＜0.05). Furthermore, serum level of CEA was obviously higher in the patients with distant metastasis than those without (P＝0.041). The serum levels of CEA and SCC-Ag were increased along with more metastases the patients had (P＜0.05). And the expression of CEA was remarkably higher in those at the stage of Ⅳb than at the stage of Ⅰ, Ⅱ, Ⅲ and Ⅳa (P＝0.046). Conclusions Serum tumor markers, CEA, CA199 and CA125 are valuable in the diagnosis of late-stage adenocarcinoma of the lung, and SCC-Ag and CYFRA21-1 are more valuable for the diagnosis of squamous cell carcinoma. There is no correlation of the five tumor markers with gender and age. Serum level of CEA is positively correlated to clinical stages of lung cancer. Namely, high level predicts an advanced stage, but low level contributes little to staging. High levels of CEA and SCC-Ag may indicate multiple metastatic sites.