Objective:To observe the efficacy and safety of anlotinib combined with chemotherapy in patients with advanced non-small cell lung cancer (NSCLC) who failed second-line chemotherapy.Methods:A retrospective analysis was performed on 80 patients with advanced NSCLC who had failed second-line chemotherapy admitted in the Department of Oncology of Chaohu Hospital of Anhui Medical University from January 2017 to October 2019, and the patients were divided into control group ( n=36) and observation group ( n=44) according to the different treatment regimens. The control group was given pemetrexed + cisplatin, and the observation group adopted pemetrexed + cisplatin + anlotinib. The objective response rate (ORR), disease control rate (DCR), median progression-free survival (PFS), overall survival (OS), changes in levels of serum vascular endothelial growth factor (VEGF), carcinoembryonic antigen (CEA) and carbohydrate antigen 199 (CA199) and treatment-related adverse reactions were compared between the two groups. Results:After 2 cycles of treatment, the ORR in the control group and observation group were 5.56% (2/36) and 18.18% (8/44), with no statistically significant difference ( χ2=1.85, P=0.174). The DCR in the two groups were 58.33% (21/36) and 81.82% (36/44), and the DCR in the observation group was significantly higher than that in the control group, with a statistically significant difference ( χ2=5.33, P=0.021). The median PFS in the two groups were 4.0 months and 6.0 months, and the median PFS in the observation group was longer than that in the control group, with a statistically significant difference ( χ2=28.47, P<0.001). The median OS in the two groups were 13.0 months and 14.8 months, with no statistically significant difference ( χ2=1.56, P=0.212). The levels of serum VEGF [(21.72±5.42) ng/L vs. (36.97±7.53) ng/L, t=14.13, P<0.001; (16.61±4.14) ng/L vs. (38.85±8.61) ng/L, t=23.09, P<0.001], CEA [(4.91±1.58) ng/ml vs. (6.62±2.84) ng/ml, t=4.64, P<0.001; (3.07±1.32) ng/ml vs. (7.08±3.31) ng/ml, t=11.50, P<0.001] and CA199 [(16.83±5.23) U/ml vs. (20.95±7.94) U/ml, t=3.75, P<0.001; (13.37±5.85) U/ml vs. (21.66±8.72) U/ml, t=7.55, P<0.001] in the control group and observation group after 2 cycles of treatment were significantly decreased compared with those before treatment, and the levels of serum VEGF, CEA and CA199 in the observation group were significantly lower than those in the control group ( t=4.78, P<0.001; t=5.68, P<0.001; t=2.76, P=0.007). The incidence of elevated blood pressure in the observation group was significantly higher than that in the control group [25.00% (11/44) vs. 2.78% (1/36), χ2=7.67, P=0.006]. Conclusion:Pemetrexed + cisplatin + anlotinib regimen for patients with advanced NSCLC who failed second-line chemotherapy can improve DCR, prolong PFS and improve the levels of serum tumor-related markers, with controllable adverse reactions.