Objective: To investigate the efficacy and safety of anlotinib as third-line and further-line treatment for advanced non-small cell lung cancer (NSCLC). Methods: A total of 25 patients with advanced NSCLC who received anlotinib for the third-line and further-line treatment from October 2018 to April 2020 at Department of Oncology of our hospital, were collected. The clinical data were analyzed retrospectively to investigate the short-term efficacy, survival, adverse events and prognostic factors of anlotinib. Results: The objective response rate (ORR) was 12% (2/25), and the disease control rate (DCR) was 80% (20/25). The median progression free survival (mPFS) was 4.00 months (95%CI: 2.78 to 5.22), and the median overall survival (mOS) was 6.50 months (95%CI: 4.05 to 8.95). Additionally, we also found that the mOS of adenocarcinoma patients was longer than that of squamous cell carcinoma patients. Univariate analysis showed that gender, age, pathology, Eastern Cooperative Oncology Group (ECOG) score, surgery, radiotherapy, history of smoking, and epidermal growth factor receptor (EGFR) mutation were not significantly correlated with OS (all P>0.05). Conclusion: Anlotinib showed favorable efficacy and safety in third-line or further-line treatment of NSCLC. To a certain extent, it can control the disease progression and prolong the survival time of NSCLC patients.