The Study Of FOXD1 Expression And Prognosis In Head And Neck Squamous Carcinoma Based On Bioinformatics

Background: Head and Neck Cancer is a common malignant tumor with high morbidity and mortality,the vast majority of them are squamous cell carcinoma.The difficulty in precision therapy for patients with head and neck squamous carcinoma(HNSC)is due to the high heterogeneity of tumor site and stage.It is very urgent to find some specific biomarkers by elucidating the occurrence and development mechanism of its molecular biology for accurate clinical treatment.Forkhead box D1(FOXD1),located on 5q13.2,is a new member of FOX transcription factor family.FOXD1 has been demonstrated multi-level roles during development,adult physiology and several diseases’ pathogenesis.However,little is known about the role of FOXD1 in the progression of HNSC.Methods: Firstly,we used online bioinformatics analysis tools to predict the expression of FOXD1 in a variety of tumors,the interacting proteins,and the functions of related genes.The expression difference of FOXD1 in HNSC and tissues adjacent to tumor was analyzed by sequencing and CHIP data from TCGA database and GEO database,and verified in cell lines and clinical samples.We also analyzed the relationship between FOXD1 expression and clinical parameters.And then,we evaluated the prognostic value of FOXD1.Moreover,the relationship between FOXD1 expression and tumor microenvironment,and also immune cell infiltration was evaluated using Estimate and CIBERSORT algorithms,respectively.Finally,gene enrichment analysis(GSEA)was used to predict the cell signaling pathways and biological functions associated with FOXD1.Results: The expression of FOXD1 in HNSC was higher than that in tissues adjacent to tumor predicted by Timer,an online bioinformatics analysis tool.The proteins interacting with FOXD1 were predicted using Gene MANIA online analysis tools.The online analysis tool of Linked Omics was used to predict the genes related to the expression of FOXD1,and the prediction of these genes through the STRING database showed that these genes were involved in the biological functions and cell signaling pathways related to the occurrence and development of cancer.Through the analysis of the TCGA datasets,the FOXD1 expression is higher in HNSC(P<0.001).11 GEO datasets were collected and analyzed,the datasets of GSE6631、GSE19089、GSE25099、GSE30784、GSE31056、GSE74530、GSE78060、GSE85195 have been found high FOXD1 expression in HNSC tissues compared with the tissues adjacent to tumor(P <0.05).FOXD1 expression was significantly associated with tumor site(P=0.010)and T stage(P=0.041).Then we validated in cell lines and clinical samples,we found that the expression of FOXD1 in oral squamous cell lines and clinical samples was higher than that in the normal oral cell lines and tissues adjacent to oral carcinoma.Kaplan-Meier survival curves showed that HNSC patients with low expression of FOXD1 in the TCGA dataset had higher overall survival than those with high expression of FOXD1(P<0.001).Univariate and multivariate Cox regression analyses showed that FOXD1 expression was an independent prognostic factor for OS(P=0.001).By the ESTIMATE algorithm,we found that the stroma(P=0.026)and immune scores(P=0.020)were significantly reduced in the group of high expression of FOXD1.Using CIBERSORT algorithm,we found that the expression of FOXD1 was negatively correlated with the proportion of naive B cells(r=-0.180,P<0.001)and plasma cells(r=-0.133,P=0.026),and positively correlated with the proportion of activated mast cells(r=0.152,P=0.002).GSEA predicted that FOXD1 was mainly involved in cancer-related signaling pathway including Bladder cancer,P53 signaling pathways,Cell cycle,Focal adhesion,Renal cell carcinoma.Metabolismrelated pathways including Protein folding,Regulation of anoikis,Positive regulation of intracellular protein transport,Regulation of cyclin dependent protein kinase activity and Nuclear envelope organization.Conclusions: Upregulation of FOXD1 is a potential prognostic marker for HNSC and is correlated with HNSC location and T stage.FOXD1 plays an important role in tumor microenvironment and immune cell infiltration.