Biomarker For Oral Squamous Cell Carcinoma Research Articles

Glycolysis-Related Gene Analyses Indicate That DEPDC1 Promotes the Malignant Progression of Oral Squamous Cell Carcinoma via the WNT/β-Catenin Signaling Pathway.

Increasing evidence suggests that aerobic glycolysis is related to the progression of oral squamous cell carcinoma (OSCC). Hence, we focused on glycolysis-related gene sets to screen for potential therapeutic targets for OSCC. The expression profiles of OSCC samples and normal controls were obtained from The Cancer Genome Atlas (TCGA). Then, the differentially expressed gene sets were selected from the official GSEA website following extraction of the differentially expressed core genes (DECGs). Subsequently, we tried to build a risk model on the basis of DECGs to predict the prognosis of OSCC patients via Cox regression analysis. Furthermore, crucial glycolysis-related genes were selected to explore their biological roles in OSCC. Two active glycolysis-related pathways were acquired and 66 DECGs were identified. Univariate Cox regression analysis showed that six genes, including HMMR, STC2, DDIT4, DEPDC1, SLC16A3, and AURKA, might be potential prognostic factors. Subsequently, a risk formula consisting of DEPDC1, DDIT4, and SLC16A3 was established on basis of the six molecules. Furthermore, DEPDC1 was proven to be related to advanced stage cancer and lymph node metastasis. Moreover, functional experiments suggested that DEPDC1 promoted the aerobic glycolysis, migration, and invasion of OSCC via the WNT/β-catenin pathway. The risk score according to glycolysis-related gene expression might be an independent prognostic factor in OSCC. In addition, DEPDC1 was identified as playing a carcinogenic role in OSCC progression, suggesting that DEPDC1 might be a novel biomarker and therapeutic target for OSCC.

Identification of a polyamine-related signature and six novel prognostic biomarkers in oral squamous cell carcinoma.

Elevated polyamine levels are required for tumor transformation and development; however, expression patterns of polyamines and their diagnostic potential have not been investigated in oral squamous cell carcinoma (OSCC), and its impact on prognosis has yet to be determined. A total of 440 OSCC samples and clinical data were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). Consensus clustering was conducted to classify OSCC patients into two subgroups based on the expression of the 17 polyamine regulators. Polyamine-related differentially expressed genes (PARDEGs) among distinct polyamine clusters were determined. To create a prognostic model, PARDEGs were examined in the training cohorts using univariate-Lasso-multivariate Cox regression analyses. Six prognostic genes, namely, "CKS2," "RIMS3," "TRAC," "FMOD," CALML5," and "SPINK7," were identified and applied to develop a predictive model for OSCC. According to the median risk score, the patients were split into high-risk and low-risk groups. The predictive performance of the six gene models was proven by the ROC curve analysis of the training and validation cohorts. Kaplan-Meier curves revealed that the high-risk group had poorer prognosis. Furthermore, the low-risk group was more susceptible to four chemotherapy drugs according to the IC50 of the samples computed by the "pRRophetic" package. The correlation between the risk scores and the proportion of immune cells was calculated. Meanwhile, the tumor mutational burden (TMB) value of the high-risk group was higher. Real-time quantitative polymerase chain reaction was applied to verify the genes constructing the model. The possible connections of the six genes with various immune cell infiltration and therapeutic markers were anticipated. In conclusion, we identified a polyamine-related prognostic signature, and six novel biomarkers in OSCC, which may provide insights to identify new treatment targets for OSCC.

NCBP2 and TFRC are novel prognostic biomarkers in oral squamous cell carcinoma

There are few prognostic biomarkers and targeted therapeutics currently in use for the clinical management of oral squamous cell carcinoma (OSCC) and patient outcomes remain poor in this disease. A majority of mutations in OSCC are loss-of-function events in tumour suppressor genes that are refractory to conventional modes of targeting. Interestingly, the chromosomal segment 3q22-3q29 is amplified in many epithelial cancers, including OSCC. We hypothesized that some of the 468 genes located on 3q22-3q29 might be drivers of oral carcinogenesis and could be exploited as potential prognostic biomarkers and therapeutic targets. Our integrative analysis of copy number variation (CNV), gene expression and clinical data from The Cancer Genome Atlas (TCGA), identified two candidate genes: NCBP2, TFRC, whose expression positively correlates with worse overall survival (OS) in HPV-negative OSCC patients. Expression of NCBP2 and TFRC is significantly higher in tumour cells compared to most normal human tissues. High NCBP2 and TFRC protein abundance is associated with worse overall, disease-specific survival, and progression-free interval in an in-house cohort of HPV-negative OSCC patients. Finally, due to a lack of evidence for the role of NCBP2 in carcinogenesis, we tested if modulating NCBP2 levels in human OSCC cell lines affected their carcinogenic behaviour. We found that NCBP2 depletion reduced OSCC cell proliferation, migration, and invasion. Differential expression analysis revealed the upregulation of several tumour-promoting genes in patients with high NCBP2 expression. We thus propose both NCBP2 and TFRC as novel prognostic and potentially therapeutic biomarkers for HPV-negative OSCC.

Identification of a CircRNA-miRNA-mRNA Network and Integrated Analysis of Immune Infiltration in Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is a highly invasive type of head and neck cancer. Circular RNA (circRNA) acts as a competing endogenous RNA (ceRNA) and involves in pathogenesis of many diseases. However, the circRNA-miRNA-mRNA network and relationship between ceRNA and immune infiltration in OSCC remain unknown. In this study, we established a ceRNA network, including 89 circRNAs, 43 miRNAs and 223 mRNAs, and found that 233 genes are mainly related to malignant signalling pathways (including "Integrin family cell surface interactions" and "Epithelial-to-mesenchymal transition" pathways) and five potential biomarkers (SLC20A1, PITX2, hsa-mir-135b, hsa-mir-377 and hsa-let-7c). Meanwhile, we established a prognostic model based on clinical risk, and revealed the relationship between immune infiltrating cells and biomarkers in OSCC. Taken together, our study is helpful to reveal the pathogenesis of oral squamous cell carcinoma.

Vascular Endothelial Growth Factor A (VEGF-A) and Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) as Potential Biomarkers for Oral Squamous Cell Carcinoma: A Sri Lankan Study.

The incidence of oral squamous cell carcinoma (OSCC) is very high in South Asia and Vascular endothelial growth factor (VEGF) is one of the key factors essential for cancer growth. The importance of VEGF-A and VEGF Receptor 2(VEGFR-2) in oral cancer pathophysiology is yet to be decided. Vascular Endothelial Growth Factor A (VEGF-A) is the main factor concerned in angiogenesis in tumors, but its role in Oral Squamous Cell Carcinoma (OSCC) is still debatable. Our study aimed to determine the role of VEGF-A and VEGFR-2 in OSCC. Blood from 30 patients with primary OSCC and 1:1 age-sex-matched controls was subjected to qPCR and ELISA to detect VEGF-A gene expression and serum level. Tumors of the 30 patients were investigated for VEGF Receptor-2 (VEGFR-2) expression and were analyzed using Image J software version 1.52 for DAB percentage (DAB-P) area and optical density (OD). VEGF-A relative gene expression among patients was 2.43-fold higher compared to the healthy control group. Well-differentiated had a 1.98-fold increment, while poorly differentiated had a 3.58-fold increment. Serum VEGF-A was significantly elevated among the patients compared to controls (458.7 vs 253.2, p=0.0225). Poorly differentiated had a higher serum VEGF concentration (1262.0±354.7pg/ml) compared with other two. Mean VEGFR-2 DAB-P level in OSCC was 42.41±5.61(p=0.15). Well-differentiated had a DAB-P of 41.20±5.32 while poorly differentiated had DAB-P 46.21±3.78. The mean OD in OSCC was 0.54±0.16. VEGFR-2 OD in well and poorly differentiated OSCC were 0.48±0.12 and 0.68±0.17, respectively. VEGF-A gene expression, serum levels, and tissue VEGFR-2 levels correlated linearly with the stage and grade of the tumor. This study justifies the value of VEGF-A as a potential biomarker in OSCC in early detection of OSCC. More studies are needed to accept the use of VEGF-A.

MICRORNA-31 AS A POTENTIAL THERAPEUTIC BIOMARKER FORORAL SQUAMOUS CELL CARCINOMA: CURRENT EVIDENCE AND FUTURE PROSPECTS.

Dear Editor,
 Cancer is a foremost death cause in each country and a substantial hindrance to improving life expectancy, accounting for more than 10 million fatalities in 2020, or around one in every six fatalities [1]. Oral cancer is among the leading causes of cancer-related mortality in developing countries. Oral squamous cell carcinoma (OSCC) is a widespread type of malignancy associated with tobacco usage and human papilloma virus infection. The majority of people are diagnosed with cancer at an advanced stage, the 5-year survival rate for OSCC patients is only around 50%. Despite biological and technological advances, in recent decades, the prognosis for OSCC has not improved, and its occurrence has increased. Identification of biomarkers that can be utilized to establish an early diagnosis or predict tumor growth is enabled by determining which molecular pathways are disrupted [2].
 MicroRNAs, which can serve as both oncogenes and tumor-inhibiting factors, are misaligned in a wide range of cancers [3]. These are short, noncoding RNAs that affect protein expression following transcription at the post-transcriptional level. MicroRNAs play a vital role in proliferation, differentiation, apoptosis, immune response, and angiogenesis, among other things in the pathogenesis of cancer. Multiple researchers have documented that specific microRNAs are frequently down regulated in cancers, implying that they operate as tumor suppressors by targeting many oncogenes. According to research conducted by O’Brien et al, altering levels of microRNAs expression can improve chemotherapeutic efficacy [4]. In recent decades, several researches have linked metastasis, survival, and recurrence of OSCC patients to miRNA expression [5].
 MicroRNA-31, a gene on chromosome 9p21.3, was one of the first mammalian microRNAs discovered. Several studies have been conducted to evaluate the functions of miR-31 in carcinogenesis and the progression of OSCC. A prior clinical investigation conducted by Liu et al. [6] revealed that miR-31 levels in plasma were considerably increased in patients with OSCC when compared to age and gender-matched control participants. Several studies show that miR-31 performs a role in cancer etiology and aggressiveness via altering target genes. According to reports, miR-31 regulates a number of genes, including fibronectin type III domain containing 5, special AT-rich sequence-binding protein-2, large tumor suppressor kinase 2 (LATS2), tensin 1, AT-rich interaction domain 1A, and hypoxia-inducible factor-1 [7]. The concentration of miR-31 expression in serum, urine, saliva and organ tissue can be utilized as a diagnostic and predictive biomarker for a variety of malignancies [8].
 A recent investigation by Chou et al. [9] found that miR-31 levels were significantly raised in OSCC cells and tumor tissues, and that the miR-31 gene locus was necessary to initiate oncogenesis in OSCC. Upregulation of miR-31 contributes to the formation of OSCC by interacting with certain genes and signaling pathways that impact cancer cell cycle, epithelial-mesenchymal transition and cell proliferation. miR-31 forms a complex network with direct target genes (such as RHOA, FIH, ACOX1, VEGF, SIRT3, LATS2, KANK1, and NUMB) and initiates several signaling pathways including the ERK-MMP9 cascade, the Hippo pathway, Wnt signaling, and the MCT1/MCT4 regulatory cascade [10]. MiR-31 interacts with a number of proteins and pathways that are crucial in OSCC and it is overexpressed in plasma, saliva, and OSCC tumor tissue. However, more research is needed to fully comprehend the molecular mechanisms by which miR-31 drives OSCC malignancy, paving the way for the development of miR-31 based diagnostic, prognostic, and predictive biomarkers for OSCC.

A novel panel of clinically relevant miRNAs signature accurately differentiates oral cancer from normal mucosa.

Although a considerable body of knowledge has been accumulated regarding the early diagnosis and treatment of oral squamous cell carcinoma (OSCC), its survival rates have not improved over the last decades. Thus, deciphering the molecular mechanisms governing oral cancer will support the development of even better diagnostic and therapeutic strategies. Previous studies have linked aberrantly expressed microRNAs (miRNAs) with the development of OSCC. We combined bioinformatical and molecular methods to identify miRNAs with possible clinical significance as biomarkers in OSCC. A set of 10 miRNAs were selected via an in silico approach by analysing the 3'untranslated regions (3'UTRs) of cancer-related mRNAs such as FLRT2, NTRK3, and SLC8A1, TFCP2L1 and etc. RT-qPCR was used to compare the expression of in silico identified miRNAs in OSCC and normal tissues (n=32). Among the screened miRNAs, miR-21-5p (p < 0.0001), miR-93-5p (p < 0.0197), miR-146b-5p (p <0.0012), miR-155-5p (p < 0.0001), miR-182-5p (p < 0.0001) were significantly overexpressed, whereas miR-133b (p < 0.05) was significantly downregulated in OSCC tissues, a scenario confirmed in two additional OSCC validation cohorts: Regina Elena National Cancer Institute (IRE cohort, N=74) and The Cancer Genome Atlas Data Portal (TCGA cohort, N=354). Initial stage tumors (T1, T2) expressed significantly higher levels of miR-133b (p < 0.0004) compared to more advanced ones (T3, T4). Also, we identified miR-93-5p (p < 0.0003), miR-133b (p < 0.0017) and miR-155-5p (p < 0.0004) as correlated with HPV-induced OSCC. The high expression of these 6 miRNAs as a signature predicted shorter disease-free survival (DFS) and could efficiently distinguish OSCC cases from healthy controls with areas under the curve (AUC) of 0.91 with sensitivity and specificity of 0.98 and 0.6, respectively. Further target identification analysis revealed enrichment of genes involved in FOXO, longevity, glycan biosynthesis and p53 cancer-related signaling pathways. Also, the selected targets were underexpressed in OSCC tissues and showed clinical significance related to overall survival (OS) and DFS. Our results demonstrate that a novel panel consisting of miR-21-5p, miR-93-5p, miR-133b, miR-146b-5p, miR-155-5p and miR-182-5p could be used as OSCC-specific molecular signature with diagnostic and prognostic significance related to OS and DFS.

Silencing of Tropomyosin 1 suppresses the proliferation, invasion and metastasis of oral squamous cell carcinoma in vitro

We focused on the expression of Tropomyosin 1 (TPM1) in oral squamous cell carcinoma (OSCC) on the basis of proteomic differential display analysis data. TPM1 is an actin binding and microfilament-associated protein, which is thought to affect the tumor behavior of various malignancies. In this study, we examined the role of TPM1 in proliferation, invasion and metastasis of OSCC cells in vitro as we tried to understand its usefulness as a potential biomarker for OSCC. Human OSCC cell lines were transfected by TPM1-siRNA. MTT assay, cell migration assay and wound healing assay were carried out to understand the characteristics of the TPM1-siRNA-transfected cells. Our data showed that suppression of TPM1 increased the proliferative ability, wound healing ability, and migratory ability of OSCC cells. These findings suggest that low expression of TPM1 could contribute to cancer cell invasion and metastasis in OSCC patients, and that TPM1 may have potential as a useful biomarker for OSCC. Availability of data and materialsAll data generated or analyzed during the present study are included in this published article.

Salivary Biomarkers in Oral Squamous Cell Carcinoma: A Proteomic Overview.

Oral squamous cell carcinoma (OSCC) is one of the most frequent cancers worldwide. Endoscopic methods may be useful in the evaluation of oral injuries even though the diagnostic gold standard is a biopsy. Targeted screenings could be considered the best way to prevent the occurrence of oral cancer. Aimed to elucidate the potential identification of specific biomarkers of OSCC, the use of saliva is convenient and noninvasive. Many studies reported more than a hundred putative saliva biomarkers for OSCC, and proteogenomic approaches were fundamental to disclosing this issue. Relevant literature published in the last few years was systematically searched on PubMed and we focused on articles about the use and study of salivary biomarkers in the diagnostics of head and neck cancer (n = 110). Thereafter, we performed a selection focusing on diagnosis with salivary proteomics in OSCC (n = 8). Saliva proteomics can be a source of biomarkers for OSCC. We reviewed literature of biomarker proteins in saliva that could also be evaluated as probable targets for non-invasive screening of oral neoplasm such as cytokines, matrix metalloproteinases, and acute-phase response proteins. The measurement of salivary biomarkers is a highly hopeful technique for the diagnosis of OSCC. Proteogenomic approaches could permit an accurate and early diagnosis of OSCC. This review seeks to generate an up-to-date view on translational OSCC issues by raising awareness of researchers, physicians, and surgeons. Renewed clinical studies, which will validate the sensitivity and specificity of salivary biomarkers, are necessary to translate these results into possible strategies for early diagnosis of OSCC, thus improving patient outcomes.

The Occurrence of Activated Leukocyte Cell Adhesion Molecule (ALCAM) and Its Predictive Factors in Patients with Oral Squamous Cell Carcinoma.

To determine the occurrence of Activated Leukocyte Cell Adhesion Molecule (ALCAM) and its predictive factors in patients with oral squamous cell carcinoma (OSCC). This cross sectional study was concocted on 102 patients with OSCC referred to Imam Khomeini Hospital of Tehran during 1997-2015. The data collection tool a checklist consisted of demographic and pathologic (lymph node involvement, differentiation, tumor size and tumor location) characteristics which extracted from patients' medical records. To evaluate ALCAM, a new sample of tumor tissue was prepared from archive. Finally, the multivariable logistic regression model was used to determine the predictive factors of ALCAM by STATA14. the number (%) of men and women were 70 (68.6) and 32 (31.4%), respectively. The mean age (S.D) of participants was 61.7 (15.6) years. Of the total samples, 32 (38.2), 19 (18.6), 36 (35.3) and 8 (7.8%) samples were related to the tongue, oral mucosa, skin and lips, respectively. More than half of the tumors had good differentiation and lymph node involvement and 74.5% were ≥20 mm. Also, 79.41% of the samples were positive for the overall incidence of ALCAM. The most important predictors of the overall incidence of ALCAM were tumor size (OR: 3.46, 95% CI: 1.71 - 7.01) and tumor location (OR: 3, 95% CI: 1.03 - 8.72). Similarly, for incidence of cytoplasmic ALCAM were age (OR: 2.56, 95% CI: 1.38 - 4.76) and location of the tumor (OR: 3.23, 95% CI: 1.08 - 9.64). However, the only predictor of membranous ALCAM incidence was lymph node involvement (OR: 0.36, 95% CI: 0.19 - 0.66). The results of our study suggest preliminary evidence for the potential clinical application of ALCAM as a prognostic biomarker for OSCC which may be the basis for future clinical application, however further studies are recommended.

Unravelling the roles of Autophagy in OSCC: A renewed perspective from mechanisms to potential applications.

Oral squamous cell carcinoma (OSCC) is associated with a low survival rate and a high disability rate, making it a serious health burden, particularly in Southeast Asian countries. Therefore, improvements in the diagnosis, treatment, and prognosis prediction of OSCC are highly warranted. Autophagy has a significant impact on cancer development. Studies on autophagy in various human cancers have made outstanding contributions; however, the relationship between autophagy and OSCC remains to be explored. This review highlights the roles of autophagy in OSCC and discusses the relationship between autophagy and Epithelial–mesenchymal transition. Considering the lack of OSCC biomarkers, we focus on the studies involving OSCC-related bioinformatics analysis and molecular targets. Based on some classical targets, we summarize several key autophagy-related biomarkers with a considerable potential for clinical application, which may become the hotspot of OSCC research. In conclusion, we elaborate on the interrelationship between autophagy and OSCC and highlight the shortcomings of current studies to provide insights into the potential clinical strategies.

TREM2 Is Associated with Advanced Stages and Inferior Prognosis in Oral Squamous Cell Carcinoma.

Simple SummaryTriggering receptor expressed on myeloid cells 2 (TREM2) is an appealing candidate for immunotherapy. However, results regarding its role in multiple cancers are rare or even contradictory. We showed that TREM2 expression in immune cells, among which were macrophages in peripheral blood mononuclear cells and tumor-associated macrophages, was associated with advanced tumor stages and reduced survival rates in oral squamous cell carcinoma (OSCC) patients. Our findings implicate that TREM2 could serve as an effective target for prognostic and therapeutic approaches in OSCC patients. Anti-TREM2 immunotherapy might greatly improve patients’ prognosis. High levels of sTREM2 were associated with advanced UICC stages, indicating that sTREM2 might be a biomarker in OSCC patients.Triggering receptor expressed on myeloid cells 2 (TREM2) is suggested to hamper antitumor immune response in multiple cancers. However, the role of TREM2 in oral squamous cell carcinoma (OSCC) and its expression in tumor-associated macrophages (TAMs) are unknown. In this study, TREM2 expression was analyzed in the primary tumors and corresponding lymph-node metastases of OSCC patients via immunohistochemistry on tissue microarrays. Human peripheral blood mononuclear cells (PBMCs) and single-cell suspensions of tumor and healthy adjacent tissues were analyzed for the presence of TREM2+ macrophages and TAMs using flow cytometry. The serum levels of soluble TREM2 (sTREM2) were quantified using an enzyme-linked immunosorbent assay. High TREM2 expression was associated with advanced UICC stages (Spearman’s rank correlation (SRC), p = 0.04) and significantly reduced survival rates in primary tumors (multivariate Cox regression, progression-free survival: hazard ratio (HR) of 2.548, 95% confidence interval (CI) of 1.089–5.964, p = 0.028; overall survival: HR of 2.17, 95% CI of 1.021–4.613, p = 0.044). TREM2 expression was significantly increased in the PBMCs of OSCC patients in UICC stage IV compared with healthy controls (ANOVA, p < 0.05). The serum levels of sTREM2 were higher in advanced UICC stages, but they narrowly missed significance (SRC, p = 0.059). We demonstrated that TREM2 was multi-factorially associated with advanced stages and inferior prognosis in OSCC patients and that it could serve as a prognostic biomarker in OSCC patients. Targeting TREM2 has the potential to reshape the local and systemic immune landscape for the potential enhancement of patients’ prognosis.

Diagnostic Strategies for Early Detection of Oral Squamous Cell Carcinoma: A Review Article

Background: Oral squamous cell carcinoma (OSCC) is the sixth most common cancer with high morbidity and mortality rates. Late diagnosis and high incidence rate of OSCC have become global healthcare issues. The purpose of this review was to provide an overview of the diagnostic and prognostic biomarkers in OSCC. Methods: A literature search in PubMed, ScienceDirect, and Google Scholar was performed. The keywords "early detection", "oral cancer", and "oral squamous cell carcinoma" were searched in the title and abstract of the articles published in English from 2000 to mid-2021. The full texts of 250 articles were retrieved and only 63 articles met the inclusion criteria. Results: In summary, all selected papers discussed the importance of early detection along with different factors and techniques to detect oral cancer. The biomarkers were divided into three groups as follows: salivary biomarkers, circulating biomarkers, and tissue biomarkers. Conclusions: In this review article, salivary biomarkers along with the circulating and tissue biomarkers were reviewed. Besides, some detection techniques were explained.

Identifying Drug Targets of Oral Squamous Cell Carcinoma through a Systems Biology Method and Genome-Wide Microarray Data for Drug Discovery by Deep Learning and Drug Design Specifications.

In this study, we provide a systems biology method to investigate the carcinogenic mechanism of oral squamous cell carcinoma (OSCC) in order to identify some important biomarkers as drug targets. Further, a systematic drug discovery method with a deep neural network (DNN)-based drug–target interaction (DTI) model and drug design specifications is proposed to design a potential multiple-molecule drug for the medical treatment of OSCC before clinical trials. First, we use big database mining to construct the candidate genome-wide genetic and epigenetic network (GWGEN) including a protein–protein interaction network (PPIN) and a gene regulatory network (GRN) for OSCC and non-OSCC. In the next step, real GWGENs are identified for OSCC and non-OSCC by system identification and system order detection methods based on the OSCC and non-OSCC microarray data, respectively. Then, the principal network projection (PNP) method was used to extract core GWGENs of OSCC and non-OSCC from real GWGENs of OSCC and non-OSCC, respectively. Afterward, core signaling pathways were constructed through the annotation of KEGG pathways, and then the carcinogenic mechanism of OSCC was investigated by comparing the core signal pathways and their downstream abnormal cellular functions of OSCC and non-OSCC. Consequently, HES1, TCF, NF-κB and SP1 are identified as significant biomarkers of OSCC. In order to discover multiple molecular drugs for these significant biomarkers (drug targets) of the carcinogenic mechanism of OSCC, we trained a DNN-based drug–target interaction (DTI) model by DTI databases to predict candidate drugs for these significant biomarkers. Finally, drug design specifications such as adequate drug regulation ability, low toxicity and high sensitivity are employed to filter out the appropriate molecular drugs metformin, gefitinib and gallic-acid to combine as a potential multiple-molecule drug for the therapeutic treatment of OSCC.

DDX59-AS1 is a prognostic biomarker and correlated with immune infiltrates in OSCC.

Background: lncRNAs play a critical role in multiple steps of gene regulation associated with tumor progression. However, the engagement of DDX59-AS1, a lncRNA, remains equivocal, particularly in oral squamous cell carcinoma (OSCC). In this study, the expression of DDX59-AS1 and its association with immune infiltration were investigated, and its prognostic value in OSSC was evaluated. Methods: OSCC patients were collected from The Cancer Genome Atlas (TCGA) database. The expression of DDX59-AS1 in OSCC and healthy tissue was compared using Wilcoxon rank sum test. The relationship between DDX59-AS1 and clinicopathological features was analyzed using Logistic regression. Gene ontology (GO) terminology analysis, gene set enrichment analysis (GSEA), and single sample GSEA (ssGSEA) were utilized to interpret the enrichment pathway and functionality and to quantify the immune cell infiltration of DDX59-AS1. The correlation between survival and DDA59-AS1 was evaluated by Kaplan-Meier analysis and Cox regression. The prognostic impact of DDX59-AS1 was predicted by the nomogram based on Cox multivariate analysis. Results: High expression of DDX59-AS1 was significantly correlated with T stage, clinical stage, race, and age (p < 0.05). Multivariate survival analysis demonstrated that the high expression of DDX59-AS1 was associated with lower overall and specific survival rates. The prognosis prediction was validated by the nomogram and calibration curves. The expression of DDX59-AS1 was negatively correlated with Mast cells, Tfh, T cells, Treg, and B cells, and positively related with the Tgd infiltration level. Conclusion: DDX59-AS1 played a crucial role in the progression and prognosis of OSCC and was potentially a predictive biomarker for OSCC.

INVESTIGATION OF METHYLATION AND EXPRESSION STATUS OF PAX1 GENE VIA EPIGENOMIC PROFILING IN ORAL MALIGNANT LESIONS

Introduction: Our previously study (TUBITAK-SBAG-114S497) showed hypermethylation of the PAX1 gene in Turkish oral squamous cell carcinoma (OSCC) patients via methylation-array results. In this study, we aimed to validate its potential as an epigenetic biomarker for OSCC by analyzing the methylation status and the changes in methylation-related gene expression of the PAX1 gene in Turkish OSCC patient and healthy groups. Material and Methods: Expression and methylation levels of PAX1 gene were analyzed by Quantitative Real Time-PCR and Quantitative Methylation Specific PCR methods respectively, in tissues and body-fluid samples of 30 OSCC patients and 30 healthy individuals. Results: The methylation rates of the PAX1 gene in matched-normal tissue, tumors, serum and saliva samples of OSCC patients were 50%, 73%, 40%, and 66%, respectively. The methylation rates of the PAX1 gene in normal mucosa, serum and saliva samples of healthy individuals were found to be 6%, 0 and 10%, respectively. In the malignant group, 59% patients in whom methylation was observed in the promoter region of the PAX1 gene in the tumor tissue was found to have decreased expression levels in the tumor tissues compared to the matched-normal tissues. Conclusion: DNA hypermethylation of PAX1 gene may play a role in oral carcinogenesis and may be used as an epigenetic biomarker.&nbsp;

Expression pattern and clinical significance of beta 2-adrenergic receptor in oral squamous cell carcinoma: an emerging prognostic indicator and future therapeutic target.

Beta 2-AdrenergicReceptor (β2-AR) is significantly overexpressed in various types of malignancies, which is associated with the worst prognosis. However, the role of β2-AR in oral cancer is not well identified. The present study aimed at investigating the β2-AR gene expression and its significance in relation withthe clinicopathological features and overall survival of oral squamous cell carcinoma (OSCC) patients. Immunohistochemistry, western blot and quantitative real-time PCR techniques were used to analyze β2-AR protein and mRNA levels in a total of 65 histopathologically confirmed OSCC tissues (case group) and 65 normal tissues (control group) from the oral cavity. Out of the total of 65 OSCC tissues, 41 tissues (63.1%) exhibited high expression for β2-AR protein. Percent positivity and relative density (mean ± SD) of protein were higher in the case group ascompared to the control group (positivity 40.31 ± 3.01 vs. 20.46 ± 1.93, p < 0.001; density 2.77 ± 1.17 vs. 1.28 ± 0.37, p < 0.001). In addition, β2-AR mRNA level was also upregulated in patients comparedtothe controls (2.36 ± 1.30 vs. 1.09 ± 0.42, p < 0.001) and showed a positive correlation with immunostaining of protein in OSCC (r = 0.48, p = 0.011). High β2-AR protein expression was significantly associated with multiple risk habits (p = 0.045), histological differentiation (p = 0.013), clinical TNM stages (p = 0.014), and poor survival (p = 0.006) of patients. In the Cox proportional hazards model, β2-AR was identified as a prognostic biomarker of OSCC (p = 0.047). β2-AR protein level is identified as an independent significant prognostic factor in patients with oral carcinoma.

Prognostic Role of Combined EGFR and Tumor-Infiltrating Lymphocytes in Oral Squamous Cell Carcinoma.

BackgroundEpidermal growth factor receptor (EGFR) is well known as a general prognostic biomarker for head and neck tumors, however the specific prognostic value of EGFR in oral squamous cell carcinoma (OSCC) is controversial. Recently, the presence of tumor-infiltrating T cells has been associated with significant survival advantages in a variety of disease sites. The present study will determine if the inclusion of T cell specific markers (CD3, CD4 and CD8) would enhance the prognostic value of EGFR in OSCCs.MethodsTissue microarrays containing 146 OSCC cases were analyzed for EGFR, CD3, CD4 and CD8 expression using immunohistochemical staining. EGFR and T cell expression scores were correlated with clinicopathological parameters and survival outcomes.ResultsResults showed that EGFR expression had no impact on overall survival (OS), but EGFR-positive (EGFR+) OSCC patients demonstrated significantly worse progression free survival (PFS) compared to EGFR-negative (EGFR-) patients. Patients with CD3, CD4 and CD8-positive tumors had significantly better OS compared to CD3, CD4 and CD8-negative patients respectively, but no impact on PFS. Combined EGFR+/CD3+ expression was associated with cases with no nodal involvement and significantly more favorable OS compared to EGFR+/CD3- expression. CD3 expression had no impact on OS or PFS in EGFR- patients. Combinations of EGFR/CD8 and EGFR/CD4 expression showed no significant differences in OS or PFS among the expression groups.ConclusionAltogether these results suggest that the expression of CD3+ tumor-infiltrating T cells can enhance the prognostic value of EGFR expression and warrants further investigation as prognostic biomarkers for OSCC.

Identification of prognostic genes in oral squamous cell carcinoma microenvironment.

Numerous studies reveal the clinical significance of tumor microenvironment (TME) in multiple cancers. The association between TME in oral squamous cell carcinoma (OSCC) and clinical outcomes remains unsolved. This study aims to exhibit the TME of OSCC and identified the prognostic marker. Gene expression profile and clinical data OSCC patients were from the TCGA database. The validated stage data was from the Gene Expression Omnibus (GSE65858). Immune/stromal scores of each patient were calculated by ESTIMATE algorithm. Biological functional prediction was conducted. Prognostic genes identified by survival analysis. Nomogram and Receiver operating characteristic curves were employed to test the predicting power. TIMER database was applied to evaluated the immune infiltrates. Lower immune scores were observed in male patients (P= 0.0107) and different primary tumor sites of oral cavity with different stromal scores (P= 0.0328). The Differentially expressed genes (DEGs) were involved in immune related pathways. HGF gene (hepatocyte growth factor) was prognostic related and with a better prognostic performance when combined with clinical features (AUC=TCGA 0.638, AUC=GEO 0.714). HGF was significantly related with B cell, CD4ï⁢¼⁢T cell, CD8+T cell, macrophage, neutrophils, and dendritic cell infiltration. The current study analyzed the TME and presented immune related prognostic biomarkers for OSCC.

Poor Prognosis of Oral Squamous Cell Carcinoma Correlates With ITGA6

ObjectivesOral cancer is the ninth most common cancer worldwide and a leading cause of cancer-related death. Oral squamous cell carcinoma (OSCC) accounts for 90% of all oral cancers. Autophagy is a conserved essential catabolic process related to OSCC. The aim of this study was to elucidate diagnostic and prognostic autophagy-related biomarkers in OSCC. MethodsThe OSCC gene expression data set was obtained from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) between the OSCC samples and adjacent healthy tissues were identified by R software. The Human Autophagy Database was screened, which revealed 222 autophagy-related genes. The autophagy-related DEGs were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were applied. Protein–protein interaction network analysis was performed in the STRING database. cytoHubba in the Cytoscape software was applied to determine the top 10 hub genes. The data set of patients with OSCC from The Cancer Genome Atlas (TCGA) was used to evaluate the prognostic value of the 10 hub genes. The association between prognosis-related hub genes and immune infiltrates was explored. ResultsTwenty-seven autophagy-related DEGs were identified. The top 10 hub genes were CCL2, CDKN2A, CTSB, CTSD, CXCR4, ITGA6, MAP1LC3A, MAPK3, PARP1, and RAB11A. ITGA6 was identified as the most efficient biomarker. Receiver operating characteristic curve analysis indicated that ITGA6 had the highest diagnostic accuracy for OSCC (area under the curve = 0.925). ITGA6 expression was significantly related to immune infiltrates. ConclusionsThe autophagy-related gene ITGA6 might be an efficient diagnostic and prognostic biomarker in OSCC.