〔Abstract〕 Objective To investigate the expression of forkhead box-3 (FOXI3) in the head and neck squamous cell carcinoma （HNSC）tissue and its impact on the prognosis of the HNSC patients, and to clarify the role of FOXI3 in the development and progression of HNSC. Methods Gene expression data and clinical information of HNSC were downloaded from The Cancer Genome Atlas (TCGA). The expression of FOXI3 in HNSC was verified using TIMER 2.0, GEPIA and UALCAN online databases. WilCoxon rank sum test was used to analyze the expression differences of FOXI3 in unpaired samples and paired samples. R software ggplot2 package was used to analyze the correlation between FOXI3 expression and HNSC clinical factors in TCGA data sets. The results were verified by Cox regression analysis of HNSC patients with prognostic information in TCGA, Kaplan Meier and GEPIA databases. R software was used for univariate and multivariate Cox analysis. GeneMania and STRING database were used to identify genes and proteins interacting with FOXI3, and clusterProfiler package of R software was used to enrich and analyze genes interacting with FOXI3. TIMER 2.0 was used to analyze the correlation between the expression of FOXI3 in HNSC and the invasion level of cancer-associated fibroblasts. The ssGSEA algorithm of R software GSVA package was used to estimate the correlation between FOXI3 expression and immune infiltration in HNSC patients in TCGA database. Results The expression of FOXI3 in HNSC tissues was significantly higher than that in normal tissues (P < 0.001). The overall survival (OS) and disease-specific survival (DSS) of HNSC patients with increased FOXI3 gene expression were worse than those of the low-expression group, and the differences were statistically significant (P < 0.05). Univariate Cox regression showed that FOXI3 was an independent prognostic factor. GeneMania database and STRING database showed that there were many genes associated with FOXI3, and enrichment analysis showed that they were involved in complex cellular components, molecular functions, biological processes and signaling pathways. Based on MCPCOUNTER, TIDE and EPIC algorithms, the expression of FOXI3 in HNSC was positively correlated with the invasion level of cancer-related fibroblasts. In HNSC, FOXI3 was positively correlated with NK CD56bright cell infiltration level, but negatively correlated with various immune cells infiltration level. Conclusion The expression of FOXI3 in HNSC tissues is higher than that in the normal tissues, and its high expression may be related to the occurrence, development and poor prognosis of HNSC. FOXI3 can be used as a candidate indicator for the diagnosis and prognosis of HNSC.