Construction of a Vasculogenic Mimicry-related RiskScore Model to Assess Patient Prognosis and Immunotherapy Response in Head and Neck Squamous Cell Carcinoma.

The functional GRHL3-filaggrin axis maintains a tumor differentiation potential and influences drug sensitivity

Glucose metabolism (GM) plays a crucial role in cancer cell proliferation, tumor growth, and survival. However, the identification of glucose metabolism-related genes (GMRGs) for effective prediction of prognosis in head and neck squamous cell carcinoma (HNSC) is still lacking. We conducted differential analysis between HNSC and Normal groups to identify differentially expressed genes (DEGs). Key module genes were obtained using weighted gene co-expression network analysis (WGCNA). Intersection analysis of DEGs, GMRGs, and key module genes identified GMRG-DEGs. Univariate and multivariate Cox regression analyses were performed to screen prognostic-associated genes. Independent prognostic analysis of clinical traits and risk scores was implemented using Cox regression. Gene set enrichment analysis (GSEA) was used to explore functional pathways and genes between high- and low-risk groups. Immune infiltration analysis compared immune cells between the two groups in HNSC samples. Drug prediction was performed using the Genomics of Drug Sensitivity in Cancer (GDSC) database. Quantitative real-time fluorescence PCR (qRT-PCR) validated the expression levels of prognosis-related genes in HNSC patients. We identified 4973 DEGs between HNSC and Normal samples. Key gene modules, represented by black and brown module genes, were identified. Intersection analysis revealed 76 GMRG-DEGs. Five prognosis-related genes (MTHFD2, CDKN2A, TPM2, MPZ, and DNMT1) were identified. A nomogram incorporating age, lymph node status (N), and risk score was constructed for survival prediction in HNSC patients. Immune infiltration analysis showed significant differences in five immune cell types (Macrophages M0, memory B cells, Monocytes, Macrophages M2, and Dendritic resting cells) between the high- and low-risk groups. GDSC database analysis identified 53 drugs with remarkable differences between the groups, including A.443654 and AG.014699. DNMT1 and MTHFD2 were up-regulated, while MPZ was down-regulated in HNSC. Our study highlights the significant association of five prognosis-related genes (MTHFD2, CDKN2A, TPM2, MPZ, and DNMT1) with HNSC. These findings provide further evidence of the crucial role of GMRGs in HNSC.

Background Head and neck squamous cell carcinoma (HNSCC) is a growing concern worldwide, due to its poor prognosis, low responsiveness to treatment, and drug resistance. Since immunotherapy effectively improves HNSCC patients' survival status, it is important to continuously explore new immune-related predictive factors to accurately predict the immune landscape and clinical outcomes of individuals suffering from HNSCC. Methods The HNSCC transcriptome profiling of RNA-sequencing data was retrieved from TCGA database, and the microarray of GSE27020 was obtained from the GEO database for validation. The differentially expressed genes (DEGs) between HNSCC and normal samples were identified by multiple test corrections in TCGA database. The univariate and multivariate Cox analyses were performed to identify proper immune-related genes (IRGs) to construct a risk model. The Cox regression coefficient was employed for calculation of the risk score (RS) of IRG signature. The median value of RS was utilized as a basis to classify individuals with HNSCC into high- and low-risk groups. The Kaplan-Meier (K-M) survival analysis and receiver operating characteristic (ROC) curves were employed for the identification of the prognostic significance and precision of the IRG signature. The signature was also evaluated based on clinical variables, predictive nomogram, mutation analysis, infiltrating immune cells, immune-related pathways, and chemotherapeutic efficacy. The protein-protein interaction (PPI) network and functional enrichment pathway investigations were utilized to explore possible potential molecular mechanisms. Finally, the hub gene's differential mRNA expression levels were evaluated by means of the Gene Expression Profiling Interactive Analysis (GEPIA), and the Human Protein Atlas (HPA) was utilized for the validation of their translational levels. Results Collectively, 1593 DEGs between HNSCC and normal samples were identified, of which 136 IRGs were differentially expressed. Then, the 136 immune-related DEGs were mostly enriched in the cytokine-related signaling pathways by GO and KEGG analyses. After that, a valuable signature based on seven genes (DKK1, GAST, IGHM, IL12RB2, SLURP1, STC2, and TNFRSF4) was designed. The HNSCC patients into the low-risk group and the high-risk group were divided by using the median RS; the HNSCC patients in the high-risk group had a worse survival than those in the low-risk group. The risk signature was verified to be an independent predictive marker for HNSCC patients. Meanwhile, the RS had the largest contribution to survival of these patients based on the predictive nomogram. In addition, the low-risk HNSCC patients exhibited significantly enriched immune cells, along with an association with high chemosensitivity. Conclusion The constructed gene signature can independently function as a predictive indicator for the clinical features of HNSCC patients. The low-risk HNSCC subjects might benefit from immunotherapy and chemotherapy.

Worldwide, Head and Neck Squamous Cell Carcinomas (HNSCC) account for about 900,000 cases and 400,000 deaths. In some settings, like Fanconi anemia (FA), patients receive curative treatments (allogeneic stem cell transplantation), only to develop HNSCC in early adulthood at a high rate of incidence. Current treatment strategies for non-FA HNSCC patients include surgery, chemotherapy and radiotherapy. However, these are not viable treatment options for FA HNSCC patients due to their low tolerance for the high toxicity levels of chemotherapy and radiation. Therefore, there is a critical need for novel and targeted therapeutic interventions for the treatment of FA HNSCC patients. B7H3, a checkpoint member of the B7 and CD28 families, is overexpressed on several solid tumors but is absent or not expressed on healthy tissues. It is a promising target for immunotherapy, and recent basket trials, particularly in the prostate cancer, have demonstrated strong clinical signals. Here we developed and tested the ability of GTB-5550, a tri-specific killer engager (TriKE) that includes a B7H3 targeting component, to direct NK cell killing to B7H3-expressing Head and Neck cancer targets. This TriKE molecule includes an NK cell engaging domain containing a humanized camelid nanobody against CD16, a camelid nanobody against B7H3 and a wild type IL-15 sequence between the two engagers. We assessed B7H3 expression by flow cytometry of wild-type HNSCC cells and a paired version with a CRISPER KO of the FANCA gene and determined that the KO had no effect on B7H3 expression. Thus, GTB-5550 activity against HNSCC should be present on both normal HNSCC and FA-HNSCC settings. NK cell responses against HNSCC lines in the presence of GTB-5550 were assessed through either flow cytometry based functional assays, to evaluate NK cell degranulation and cytokine secretion, or IncuCyte imaging assays, to directly assess target killing. NK cell degranulation and IFN-gamma production of GTB-3550-treated samples were higher compared to that of control samples treated with B7H3 single domain or IL-15 alone. GTB-5550 also induced more HNSCC target cell killing by NK cells compared to treatment with the B7H3 single domain or IL-15 alone irrespective of the FANCA gene. Ongoing experiments will evaluate functionality of GTB-5550 on FA patient samples as well as in spheroid assays. Taken together, this data shows that a GTB-5550 is able to drive NK cell activity against B7H3-expressing HNSCC cells, which presents potential for a B7H3-targeted TriKE to be used to be implemented clinically to treat HNSCC or FA-HNSCC patients. Citation Format: Melissa Khaw, Zachary Davis, Nicholas Zorko, Greg Berk, Gavin Choy, Margaret MacMillan, Martin Felices, Jeffrey S. Miller. GTB-5550 (cam16-IL15-camB7H3) trispecific killer engager (TriKE®) drives natural killer cell activation and antibody dependent cellular cytotoxicity against head and neck squamous cell carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3435.

Association of higher transient receptor potential melastatin 8 expression with higher tumor histologic grades, lymph node metastasis, risk factors, and worse survival in patients with head and neck squamous cell carcinoma

Background: Radioresistance in head and neck squamous cell carcinoma (HNSCC) patients means response failure to current treatment. In order to screen radioresistant biomarkers and mechanisms associated with HNSCC, differentially expressed genes (DEGs) associated with radioresistance in HNSCC were investigated.Methods: The HNSCC cell line with radioresistance, Hep2-R, was established and detected the radiosensitivity using MTT, colony formation assay and flow cytometry analysis. Clariom™ D chip was applied to compare DEGs between Hep2 and Hep2-R groups and build the differential gene expression profiles associated with radioresistance in HNSCC. Bioinformatic analysis were used to find biological functions and pathways that related to radioresistance in HNSCC, including cell adhesion, cytochrome P450 and drug metabolism. Gene Expression Omnibus (GEO) datasets were selected to verify DEGs between HNSCC radioresistant cells and tissues. The representation of DEGs were validated between HNSCC patients with complete response and post-operative radiation therapy failure. In addition, we evaluated the clinical prognosis of DEGs using The Cancer Genome Atlas (TCGA) database.Results: 2,360 DEGs (|Fold Change|>1.5, p < 0.05) were identified between Hep2 and Hep2-R, including 1,144 upregulated DEGs and 1,216 downregulated DEGs. They were further verified by HNSCC radioresistant cells and tissues in GEO. 13 radioresistant DEGs showed same difference in expression level between cells and tissues. By comparing 13 DEGs with HNSCC patients, upregulations of FN1, SOX4 and ETV5 were found identical with above results. Only FN1 was a prognostic indicator of HNSCC in TCGA.Conclusion: FN1 is the potential novel biomarker for predicting poor prognosis and radioresistance in HNSCC patients. Overexpression of FN1 plays an important role in the tumorigenesis, prognosis and radioresistance of HNSCC.

The fatty acid metabolism (FAM) is known to impact tumorigenesis, tumor progression and treatment resistance via enhancing lipid synthesis, storage and catabolism. However, the role of FAM in head and neck squamous cell carcinoma (HNSCC) has remained elusive. In the present study, we obtained a total of 69 differentially expressed FAM-related genes between 502 HNSCC samples and 44 normal samples from The Cancer Genome Atlas (TCGA) database. The HNSCC samples were divided into 2 clusters according to 69 differentially expressed genes (DEGs) via cluster analysis. Then DEGs in the two clusters were found, and 137 prognostic DEGs were identified by univariate analysis. Subsequently, combined with the clinical information of 546 HNSCC patients from TCGA database, a 12-gene prognostic risk model was established (FEPHX3, SPINK7, FCRLA, MASP1, ZNF541, CD5, BEST2 and ZAP70 were down-regulation, ADPRHL1, DYNC1I1, KCNG1 and LINC00460 were up-regulation) using multivariate Cox regression and LASSO regression analysis. The risk scores of 546 HNSCC samples were calculated. According to the median risk score, 546 HNSCC patients were divided into the high- and low-risk (high- and low score) groups. The Kaplan-Meier survival analysis showed that the survival time of HNSCC patients was significantly shorter in the high-risk group than that in the low-risk group (p < 0.001). The same conclusion was obtained in the Gene Expression Omnibus (GEO) dataset. After that, the multivariate Cox regression analysis indicated that the risk score was an independent factor for patients with HNSCC in the TCGA cohort. In addition, single-sample gene set enrichment analysis (ssGSEA) indicated that the level of infiltrating immune cells was relatively low in the high-risk group compared with the low-risk group. In summary, FAM-related gene expression-based risk signature could predict the prognosis of HNSCC independently.

Head and neck squamous cell carcinomas (HNSCC) are the sixth most common cancer type worldwide and result in 400,000 deaths annually. Interleukin-9 (IL-9) is a pleiotropic cytokine largely produced by CD4+ Th9 cells. IL-9 is known to function in allergic inflammatory processes and is involved in both positive and negative regulation of immune responses. The role of IL-9 in cancer, however, differs depending on the cancer type and the tumor microenvironment, harboring both tumorigenic and anti-tumorigenic effects. The role of IL-9 in HNSCC specifically is poorly understood. We begin here to elucidate the possible functions of IL-9 on the immune system in the context of HNSCC.IL-9 plasma levels were determined by ELISA and cell proliferation was measured using a metabolic absorbance assay. Kaplan-Meier Plotter was used to determine if tumor IL-9 mRNA expression affects survival probability in HNSCC patients. Immune cell infiltration was determined by a novel infiltration assay designed by our laboratory. Briefly, we co-cultured Cal27 spheroids, a 3D model made with the HNSCC cell line Cal27, with peripheral blood mononuclear cells (PBMCs) isolated from HNSCC patients. The spheroids were then dissociated and the percentage of live infiltrating PBMCs was analyzed by flow cytometry. To assess the influence of IL-9 on immune cell killing, NK and CD8+ T cell cytotoxicity was measured by co-culturing CFDA-SE-labeled HNSCC cell lines with HNSCC patient-derived NK or T cells treated with recombinant IL-9 or a neutralizing antibody to IL-9 followed by flow cytometry analysis to determine the percentage of dead target cells.IL-9 plasma levels were found to be significantly increased in HNSCC patients as compared to healthy subjects. Additionally, high tumoral mRNA expression of IL-9 was associated with decreased survival in HNSCC cohorts. IL-9 did not affect tumor cell proliferation in vitro. However, higher concentrations of IL-9 significantly decreased immune cell infiltration into Cal27 spheroids, while a neutralizing antibody to IL-9 reversed these effects. Additionally, while studies are ongoing, IL-9 may decrease NK and CD8+ T cell cytotoxicity against HNSCC cell lines.Since IL-9 did not directly enhance or inhibit HNSCC cell proliferation in vitro, we extended our studies to explore IL-9’s potential effects on the immune system. Importantly, IL-9 significantly decreased immune cell infiltration into HNSCC spheroids, suggesting that IL-9 inhibits tumor immune cell infiltration. IL-9 may also decrease immune cell cytotoxicity against HNSCC cells. Taken together, this data suggests that IL-9 negatively regulates the immune system in HNSCC, allowing for tumor growth. Future studies will focus on the effects of IL-9 on immune cell cytotoxicity and immune infiltration in vivo and will utilize single-cell transcriptional profiling to delineate the multifaceted role of IL-9 in the HNSCC immune microenvironment. Citation Format: Samantha L. Nusbaum, Maria Lehn, Vinita Takiar, Dalia El-Gamal, Trisha Wise-Draper. The role of IL-9 on NK and T cell infiltration and immune response to promote tumorigenesis in HNSCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5851.

Telomere dysfunction has been identified as a biological marker of cancer progression in several types of cancer, including Head and Neck Squamous Cell Carcinoma (HNSCC). This study aimed to characterize the telomere maintenance genes (TMG)-related signature in prognosis and treatment response in HNSCC. The transcriptome and clinical data of HNSCC were obtained from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus databases, respectively. Non-negative matrix factorization (NMF) was used to identify molecular subtypes derived from TMG. Gene set enrichment analysis (GSEA) was performed to analyze the differentially expressed pathways between subtypes, and a risk score model derived from TMG was established. Kaplan-Meier survival analysis was used to evaluate inter-group prognostic features, and the correlation between TMG-derived molecular subtypes and risk score model with immune infiltration, immunotherapy, and chemosensitivity was assessed. Two HNSCC subtypes were identified based on 59 TMG-related genes, which exhibit significant heterogeneity in prognosis, immune cell infiltration, and treatment response. Additionally, a TMG-derived risk signature containing 9 genes was developed to assess the prognosis of HNSCC patients. The signature had significant predictive ability for HNSCC prognosis and was significantly correlated with immune cell infiltration and immunotherapy response. A nomogram integrating the risk signature, N stage and radiotherapy was constructed to predict 1-, 3-, and 5-year overall survival (OS) of HNSCC patients, which had better performance than other prognostic models and included TMG-derived risk score, radiotherapy, and N stage. This study identified TMG-derived molecular subtypes in HNSCC and developed a novel prognostic score model, highlighting the potential value of TMG in HNSCC prognosis and immunotherapy.

This review article explores phosphatase and tensin homolog (PTEN)’s role in head and neck squamous cell carcinoma (HNSCC) through comprehensive expression and methylation examinations, genetic mutation investigation, and prognostic evaluation. Using the UALCAN informational collection, PTEN expression examination uncovered a critical over-expression in HNSCC cells isolated from normal control samples, proposing its role in HNSCC multiplication. Further, analysis of PTEN expression across various clinical limits has shown critical up-regulation in different cancer development stages, racial groups, gender, and age classes within the context of HNSCC patients, suggesting its major role in cancer duplication. PTEN expression was validated by utilizing the GEPIA2.0 online tool, which showed PTEN expression was particularly significantly expressed in HNSCC cancer improvement when it appeared differently from normal control samples. Accordingly, examining PTEN validation across different phases of cancer advancement showed dysregulation in each of the four phases with the most raised expression in stage I and the least expression in stage IV. Thus, this study investigated the promoter methylation level of PTEN, figuring out a basic relationship between HNSCC samples and normal control samples. Analyzing promoter methylation across various clinical limits uncovered massive variations, with specific methylation patterns seen across malignant growth stages, race groups, gender, and age groups. Overall survival and disease-free survival (OS and DFS) utilizing the KM plotter tool showed a critical relationship between PTEN expression levels in HNSCC patients, showing high PTEN expression exhibited good overall survival when showed up distinctively comparable to low PTEN expression levels. In addition, in disease-free survival (DFS) evaluation HNSCC patients showing low PTEN expression experienced great DFS relative to HNSCC patients with high PTEN expression. Moreover, to validate PTEN expression against survival, the study examined the HNSCC patients into low and high-expression groups of PTEN. In HNSCC, low PTEN expression was connected with great overall survival (OS) when it appeared contrastingly relative to the high PTEN expression. In like manner, the study found that low PTEN expression level was connected with great DFS in HNSCC when it appeared contrastingly related to the high PTEN expression group. Genetic mutation analysis via cBioPortal identifies a minimal proportion of PTEN mutations in HNSCC, predominantly in-frame mutation, missense mutation, splice mutation, truncating mutation, and structural variant, indicating their basal significance in PTEN dysregulation within HNSCC. Further investigation of PTEN molecular components and their exchange inside the HNSCC microenvironment might disclose novel roads for designated treatment and accurate medication approaches in battling this harmful disease.

BackgroundRecent research has highlighted pyroptosis as a key factor in cancer progression. This study aims to explore the association between pyroptosis-related signatures and overall survival (OS) in head and neck squamous cell carcinoma (HNSC) and develop a pyroptosis-related long non-coding RNA (lncRNA) risk model to predict prognosis and response to immunotherapy in HNSC.MethodsWe extracted expression data for 18 pyroptosis-related genes and identified lncRNA probes specific to HNSC by using datasets from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). Consensus clustering was performed to categorize HNSC patients into distinct subtypes. A six-lncRNA risk score model was constructed through univariate and least absolute shrinkage and selection operator (LASSO) Cox regression analyses. We evaluated the predictive ability of the lncRNA model for patients’ survival and immunotherapy response. Gene expression was evaluated using immunohistochemistry (IHC) and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR).ResultsOur analysis revealed two distinct pyroptosis-related subtypes in HNSC patients, Cluster A and Cluster B. Notably, patients in Cluster B exhibited significantly poorer overall survival compared to those in Cluster A. Through differential expression analysis, we identified six lncRNAs (AC002331.1, CTA-384D8.35, RP11-291B21.2, AC006262.5, RP1-27K12.2, and RP11-54H7.4) that were differentially expressed between these clusters. A 6-lncRNA risk score model was developed, which successfully stratified patients into high- and low-risk groups with distinct overall survival outcomes. Validation using RT-qPCR confirmed the differential expression of these six lncRNAs in HNSC tumor tissues compared to adjacent normal tissues, we found that the expression of CTA-384D8.35 was significantly increased in the tumor group (t=-6.203, P<0.001). Furthermore, the 6-lncRNA risk score model demonstrated a significant association with patient response to immunotherapy, with the low-risk group exhibiting a higher objective response rate to immune checkpoint blockade (ICB) therapy and longer survival compared to the high-risk group.ConclusionOur study underscores the role of pyroptosis signatures in HNSC prognosis and identifies two distinct pyroptosis subtypes with differing survival outcomes. The six-lncRNA risk score model offers a valuable tool for predicting patient prognosis and potential benefits from ICB therapy. These findings highlight the importance of pyroptosis and associated lncRNAs in the tumor microenvironment, paving the way for novel targeted therapies in HNSC.

Objectives: Head and neck squamous cell carcinoma (HNSCC) is the most common malignancy of the head and neck. However, the molecular mechanisms governing the development of HNSCC have not been fully elucidated.Materials and Methods: Differentially expressed genes (DEGs) were screened out from The Cancer Genome Atlas (TCGA) and GSE23036 datasets. Weighted gene coexpression network analysis (WGCNA) was used to reveal the correlations among genes and to search for significantly correlated gene modules. The expression levels of genes in HNSCC and normal samples according to antibody-based detected methods was assessed by utilizing the Human Protein Atlas (HPA). The impact of the selected hub genes on the prognosis of HNSCC patients was assessed by analysing immunohistochemistry (IHC) and immunofluorescence (IF) expression levels and clinical data.Results: Twenty-four genes positively correlated with tumour status and 15 genes negatively correlated with tumour status were screened out by WGCNA. PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC and were finally screened out and verified by GEPIA and HPA database analysis. Immunohistochemistry of samples collected from 175 patients with HNSCC and subsequent statistical analysis also showed that PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC, and the levels of these two factors were positively correlated. The expression and co-localization of PLAU and LAMC2 in HNSCC tissues were confirmed by double immunofluorescence labeling.Conclusions: There was a positive correlation between PLAU and LAMC2 expression in HNSCC samples, and PLAU and LAMC2 might be independent prognostic biomarkers for HNSCC.

Head and neck squamous cell carcinoma (HNSCC) is notorious for poor prognoses, and effective biomarkers are urgently needed for early diagnosis of HNSCC patients. We investigate the role of alkaline ceramidase 1 (ACER1) and its relationship with immune infiltration in HNSCC. The differential expression and clinical prognostic significance of ACER1 in HNSCC patients are explored using bioinformatics methods and verified in human HNSCC samples. Genetic mutation, DNA methylation and drug sensitivity linked with ACER1 are examined. The potential biological function of ACER1 co-expression genes is assessed, and a series of functional assays are performed on ACER1in vitro. The results comprehensively reveal a relationship between ACER1 and immune infiltration in HNSCC patients. ACER1 expression is significantly downregulated in HNSCC tissues and closely correlated with better prognoses for HNSCC patients, and this prognostic significance is determined by distinct clinical characteristics. Genetic alteration and promoter hypomethylation of ACER1 are involved in progression of HNSCC, and ACER1 expression is significantly related to several drug sensitivities. Functional analysis shows that ACER1 co-expression genes are mainly enriched in the sphingolipid signaling pathway associated with inhibition of tumorigenesis, leading to better prognoses for HNSCC patients. In vitro, ACER1 overexpression inhibits proliferation and migration, induces apoptosis, and promotes adhesion of Fadu and SCC9 cells. In addition, high ACER1 expression is closely linked with infiltration levels of immune cells, and strongly associated with biomarkers of immune cells in HNSCC, suggesting the important role of ACER1 in regulating tumor immunity in HNSCC patients. In summary, ACER1 may be a useful indicator for diagnosis and prognosis, and may regulate immune infiltration in HNSCC patients, thus promising targeted immunotherapy for HNSCC.

The prognostic value of TMB and the relationship between TMB and immune infiltration in head and neck squamous cell carcinoma: A gene expression-based study.

BackgroundHead and neck squamous cell carcinoma (HNSCC) is one of the most common cancers worldwide, exhibiting high morbidity and mortality. The prognosis of HNSCC patients has remained poor, though considerable efforts have been made to improve the treatment of this cancer. Therefore, identifying significant differentially expressed genes (DEGs) involved in HNSCC progression and exploiting them as novel biomarkers or potential therapeutic targets for HNSCC is highly valuable.MethodsOverlapping differentially expressed genes (DEGs) were screened out from three independent gene expression omnibus (GEO) datasets and subjected to GO and kyoto encyclopedia of genes and genomes pathway enrichment analyses. The protein–protein interactions network of DEGs was constructed in the STRING database, and the top ten hub genes were selected using cytoHubba. The relative expression of hub genes was detected in GEPIA, Oncomine, and human protein atlas (HPA) databases. Furthermore, the relationship of hub genes with the overall survival and disease‐free survival in HNSCC patients was investigated using the cancer genome atlas data.ResultsThe top ten hub genes (SPP1, POSTN, COL1A2, FN1, IGFBP3, APP, MMP3, MMP13, CXCL8, and CXCL12) could be utilized as potential diagnostic indicators for HNSCC. The relative levels of FN1, APP, SPP1, and POSTN could be associated with the prognosis of HNSCC patients. The mRNA expression of APP and COL1A2 was validated in HNSCC samples.ConclusionThis study identified effective and reliable molecular biomarkers for diagnosis and prognosis by integrated bioinformatics analysis, suggesting novel and essential therapeutic targets for HNSCC.