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Objective: To analyze the changes in tumor necrosis factor receptor superfamily (TNFRSF) 4 expression levels in laryngeal squamous cell carcinoma (LSCC) patients through multiple database data combined with clinical samples, and to study the role of TNFRSF4 in the occurrence and development of LSCC. Method: Based on Gene Expession Omnibus (GEO) database and The Cancer Genome Atlas (TCGA) database, the differentially expressed gene was screened out. The expression level prediction and survival analysis of TNFRSF4 were performed using Gene Expression Profiling Interactive Analysis (GEPIA) database. Western blot and qRT-PCR were performed on LSCC and adjacent tissue samples from three patients. K-M curve was drawn based on TCGA clinical data, and Cox regression analysis was performed. Finally, gene set enrichment analysis was performed to explore signaling pathways related to the role of TNFRSF4 in LSCC. Results: In the database and clinical samples, the expression of TNFRSF4 in the LSCC group was higher than that in the normal group (P＜0.05), and the survival rate of the high TNFRSF4 expression group was higher than that of the low TNFRSF4 expression group (P＜0.01). Multivariate Cox regression analysis showed that the expression level of TNFRSF4 (HR: 0.430, 95%CI: 0.229-0.806, P=0.009), gender (HR: 0.424, 95%CI: 0.204-0.882, P=0.022), lymph node staging (HR:2.010, 95%CI: 1.055-3.831, P=0.034) and distant metastasis (HR: 3.706, 95%CI: 1.152-11.922, P=0.028) collectively affect the overall survival time of patients. The results of gene set enrichment analysis showed that the most significantly correlated signaling pathways with TNFRSF4 expression included cell adhesion molecule, JAK-STAT signaling pathway, T cell receptor signaling pathway, B cell receptor signaling pathway, primary immunodeficiency, and autoimmune thyroid disease. Conclusion: High expression of TNFRSF4 may be a biomarker for good prognosis of patients with LSCC.