Abstract: Objective To observe the changes of lncRNA HOTAIR and tumor markers in different pathological stages of lung cancer, and to explore the clinical value of lncRNA HOTAIR combined with tumor markers in the diagnosis of lung cancer stages. Methods Fifty patients with lung cancer in the first affiliated Hospital of Bengbu Medical College were selected as the lung cancer group and 50 healthy volunteers as the control group. According to the T and N stages of lung cancer, 50 patients with lung cancer were divided into T1, T2, T3 and N0, N1, N2 groups. The levels of lncRNA HOTAIR in cancer and paracancerous tissues were detected by fluorescence in situ hybridization, and the levels of NSE, CEA and CYFRA21-1 in serum were detected by immunofluorescence. Results The serum levels of NSE, CEA and CYFRA21-1 in the lung cancer group were higher than those in the control group, and the expression levels of, lncRNA HOTAIR, NSE, CEA and CYFRA21-1 in the lung cancer group were significantly higher than those in the control group(all P<0.05). In the T and N stages of lung cancer, the expression levels of, lncRNA HOTAIR, NSE, CEA and CYFRA21-1 were significantly higher than those in the control group(all P<0.05). LncRNA HOTAIR concentration in paracancerous tissues was lower than that in lung cancer tissues(P<0.001), and the serum levels of NSE, CEA and CYFRA21-1 in lung cancer group were higher than those in control group(all P<0.05). The expression of lncRNA HOTAIR was positively correlated with the expression of NSE, CEA and CYFRA21-1. The expression levels of lncRNA HOTAIR in tissue and serum NSE, CEA and CYFRA21-1 in patients with lung cancer were positively correlated with T stage and N stage of lung cancer(all P<0.001), and were positively correlated with the N stage of lung cancer, and the expression levels of tissue and serum NSE, CEA, CYFRA21-1 in patients with lung cancer were positively correlated with T stage and N stage of lung cancer(all P<0.001). The sensitivity, approximate equal index and positive predictive value of combined detection were higher than those of single index. Conclusion The expression level of lncRNA HOTAIR and serum NSE, CEA, CYFRA21-1 is closely related to the pathological stage of lung cancer. lncRNA HOTAIR combined with serum tumor markers can improve the accuracy of detection of pathological stage of lung cancer.