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Objective: To study the relationship between circulating tumor cells （CTC） and serum scc-ag and keratin 19 soluble fragment（cy-fra21-1） levels in patients with non-small cell lung cancer （NSCLC）. Methods: 100 NSCLC patients admitted to our hospital from February 2017 to February 2019 were included in the study, which was referred to as NSCLC group. Another 100 patients with benign lung lesions admitted to our hospital at the same time were taken as the benign lesion group. Another 100 healthy volunteers who underwent physical examination in our hospital at the same time were selected as the control group. The CTC and serum scc-ag and cy-fra21-1 levels of the three groups were detected and compared, and the correlation between CTC and serum scc-ag and cy-fra21-1 levels was analyzed by Pearson correlation. The diagnostic efficacy of CTC and serum scc-ag and cy-fra21-1 levels in NSCLC was analyzed by ROC curve. In addition, the relationship between CTCS, serum scc-ag, cy-fra21-1 and lymph node metastasis in NSCLC patients was analyzed. Results: The levels of CTC, serum scc-ag and cy-fra21-1 in the NSCLC group and the benign lesion group were higher than those in the control group, and the levels of CTC, serum scc-ag and cy-fra21-1 in the NSCLC group were higher than those in the benign lesion group （all P<0.05）. Pearson correlation analysis showed that there was a positive correlation between CTC and serum scc-ag and cy-fra21-1 levels in NSCLC patients （r=0.642、0.601, P<0.05）. According to ROC curve analysis, the combined detection of CTC, serum scc-ag and cy-fra21-1 levels in the diagnosis of NSCLC showed that the area under the curve, the sensitivity and the specificity were all higher than those measured separately （all P<0.05）. The levels of CTC and serum scc-ag and cy-fra21-1 in patients with NSCLC lymph node metastasis were higher than those in patients without lymph node metastasis （all P<0.05）. Conclusion: There is a significant positive correlation between CTCS and serum scc-ag and cy-fra21-1 levels in NSCLC patients, and the combined detection of the three can improve the diagnostic value of NSCLC, and the above three indicators are closely related to lymph node metastasis. [ABSTRACT FROM AUTHOR]