Objective To establish a subcutaneous tumor model in nude mice using patient-derived tissues (PDTs) of lung squamous cell carcinoma (LSCC) and determine the sensitization effect of anlotinib on radiofrequency ablation (RFA)-mediated inhibition of subcutaneous LSCC growth. Methods Patient-derived LSCC tissue was collected and qPCR was used to detect the expression of the target of anlotinib. Then, the tissue was subcutaneously implanted into nude mice to form tumor tissues. Oral administration of anlotinib and RFA treatment of subcutaneous tumors were performed, and qPCR was used to detect epithelial-mesenchymal transition of LSCC. Results Subcutaneous tumor tissue in nude mice was generated from patient-derived LSCC tissues. The target of anlotinib was clearly expressed. RFA effectively killed subcutaneous LSCC tumor tissues. Anlotinib affected RFA by inhibiting EMT in tumor tissue to sensitize LSCC tissues to killing. Conclusions Anlotinib increases the killing effect of RFA on patient-derived LSCC tissues in a nude mouse model, which may facilitate the diagnosis and treatment of LSCC.