Oral Squamous Cell Carcinoma Tissue Samples Research Articles

M6A-Mediated IRS1 Regulates the Development of Oral Squamous Cell Carcinoma through p53/Line-1 Signaling.

The importance of N6-methyladenosine (m6A) modification in tumorigenesis and progression have been highlighted. This study aimed to investigate the modification of insulin receptor substrate 1 (IRS1) by m6A and its role in oral squamous cell carcinoma (OSCC). Bioinformatics was employed to predict differential genes related to epithelial-mesenchymal transition (EMT) in OSCC. Seventeen pairs of OSCC and paracancerous tissue samples were collected. The impact of IRS1 on OSCC cell growth and EMT was evaluated. The fluctuations in IRS1 enrichment and the involvement of p53/Line-1 were investigated. IRS1 was highly expressed in OSCC. IRS1 silencing decreased OSCC cell proliferation and increased apoptosis. IRS1 silencing hindered EMT by regulating related markers. IRS1 silencing upregulated p53 and downregulated Line-1 ORF1p. The p53 inhibition reversed the effects of IRS1 silencing and induced EMT in OSCC cells. Furthermore, the m6A modification of IRS1 was increased in OSCC cells. IRS1 were positively regulated by the m6A regulators methyltransferase-like 14 (METTL14) and YTH domain-containing protein 1 (YTHDC1). IRS1 bound to YTHDC1, and YTHDC1 knockdown inhibited the IRS1 nuclear export. The obesity-associated protein (FTO) negatively regulated IRS1, and FTO overexpression reversed the IRS1-induced OSCC tumor growth. m6A methylation-mediated IRS1 regulated EMT in OSCC through p53/Line-1. These findings provide potential therapeutic strategies for managing OSCC.

PAK4 inhibition augments anti-tumour effect by immunomodulation in oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is one of the most common malignant tumours, warranting novel treatments. Here, we examined the therapeutic efficacy of inhibiting p21 activated kinase 4 (PAK4) in OSCC and determined its immunomodulatory effect by focusing on the enhancement of anti-tumour effects. We examined PAK4 expression in OSCC cells and human clinical samples and analysed the proliferation and apoptosis of OSCC cells following PAK4 inhibition in vitro. We also investigated the effects of in vivo administration of a PAK4 inhibitor on immune cell distribution and T-cell immune responses in OSCC tumour-bearing mice. PAK4 was detected in all OSCC cells and OSCC tissue samples. PAK4 inhibitor reduced the proliferation of OSCC cells and induced apoptosis. PAK4 inhibitor significantly attenuated tumour growth in mouse and was associated with increased proportions of IFN-γ-producing CD8+ T-cells. Furthermore, PAK4 inhibitor increased the number of dendritic cells (DCs) and up-regulated the surface expression of various lymphocyte co-stimulatory molecules, including MHC-class I molecules, CD80, CD83, CD86, and CD40. These DCs augmented CD8+ T-cell activation upon co-culture. Our results suggest that PAK4 inhibition in OSCC can have direct anti-tumour and immunomodulatory effects, which might benefit the treatment of this malignancy.

The Immune Checkpoint BTLA in Oral Cancer: Expression Analysis and Its Correlation to Other Immune Modulators.

In oral squamous cell carcinoma (OSCC) tissues, an immunotolerant situation triggered by immune checkpoints (ICPs) can be observed. Immune checkpoint inhibitors (ICIs) against the PD1/PD-L axis are used with impressive success. However, the response rate is low and the development of acquired resistance to ICI treatment can be observed. Therefore, new treatment strategies especially involving immunological combination therapies need to be developed. The novel negative immune checkpoint BTLA has been suggested as a potential biomarker and target for antibody-based immunotherapy. Moreover, improved response rates could be displayed for tumor patients when antibodies directed against BTLA were used in combination with anti-PD1/PD-L1 therapies. The aim of the study was to check whether the immune checkpoint BTLA is overexpressed in OSCC tissues compared to healthy oral mucosa (NOM) and could be a potential diagnostic biomarker and immunological target in OSCC. In addition, correlation analyses with the expression of other checkpoints should clarify more precisely whether combination therapies are potentially useful for the treatment of OSCC. A total of 207 tissue samples divided into 2 groups were included in the study. The test group comprised 102 tissue samples of OSCC. Oral mucosal tissue from 105 healthy volunteers (NOM) served as the control group. The expression of two isoforms of BTLA (BTLA-1/2), as well as PD1, PD-L1/2 and CD96 was analyzed by RT-qPCR. Additionally, BTLA and CD96 proteins were detected by IHC. Expression levels were compared between the two groups, the relative differences were calculated, and statistical relevance was determined. Furthermore, the expression rates of the immune checkpoints were correlated to each other. BTLA expression was significantly increased in OSCC compared to NOM (pBTLA_1 = 0.003; pBTLA_2 = 0.0001, pIHC = 0.003). The expression of PD1, its ligands PD-L1 and PD-L2, as well as CD96, were also significantly increased in OSCC (p ≤ 0.001). There was a strong positive correlation between BTLA expression and that of the other checkpoints (p < 0.001; ρ ≥ 0.5). BTLA is overexpressed in OSCC and appears to be a relevant local immune checkpoint in OSCC. Thus, antibodies directed against BTLA could be potential candidates for immunotherapies, especially in combination with ICI against the PD1/PD-L axis and CD96.

Expression levels of microRNA-7110 in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a leading cause of cancer-related deaths worldwide, and it is responsible for more than 95% of head and neck cancers. Despite advancements in research and treatment, patient's survival has not significantly increased in recent years. On the other hand, microRNAs (miRNAs) are a major class of small non-coding RNAs that regulate gene expression of the target mRNAs. Thus, understanding the mechanisms behind OSCC formation and progression may lead to the identification of potential diagnostic biomarkers and therapeutic molecules for the treatment of OSCC. The aim of the current study was to analyze expression levels of miR-7110 in OSCC tissues and adjacent normal tissues as it could provide insights into its potential role in OSCC development or progression as a valuable biomarker. A total of 20 OSCC and adjacent normal tissues were collected from the Department of Oral and Maxillofacial Surgery, Saveetha Dental College and Hospitals (Chennai, India). The tissues were processed for hematoxylin and eosin staining and expression studies. The data were shown as mean±standard deviation and P<0.05 was considered statistically significant. Our histopathological observations revealed an invasive malignant epithelial neoplasm with malignant epithelial cells exhibiting features of severe epithelial dysplasia invading the connective tissue stroma as islands, strands and cords with varying degrees of differentiation. Our results have also revealed that the expression levels of miR-7110 were found to be significantly higher in OSCC samples when compared to the normal tissue. We can preliminarily conclude that based on the increased expression of miR-7110 in OSCC tissue samples, they can be used as an early diagnostic or prognostic biomarker and/or a therapeutic target for the treatment of OSCC even though more focused research in that direction is needed.

Effect of the Protein Kinase B (PKB) Gene on the Carcinogenesis of Oral Squamous Cell Carcinoma in the South Indian Population.

The most common head and neck cancer is oral squamous cell carcinoma (OSCC). It is also one of the most prevalent forms of cancer globally. The current pharmacological treatment strategy for oral cancer lacks specificity and is capable of causing various side effects. This fact highlights the increasing need for targeted therapy. Interestingly, protein kinase B (PKB), commonly referred to as the AKT serine/threonine kinase, is an oncogenic protein that controls cell development, proliferation, apoptosis, and glycogen metabolism. Thus, the present study analyzed the AKT gene expression in OSCC patient samples. A total of 25 OSCC tissue samples and normal tissue samples were collected from the patients who reported to the Department of Oral and Maxillofacial Surgery, Saveetha Dental College and Hospitals in Chennai, India. The tissues were processed for H&E staining for histopathological confirmation, and expression studies of the AKT gene were done on both healthy and proven OSCC tissue samples. The data were shown as mean ± standard deviation, and p<0.05* was considered to be statistically significant. The quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis revealed that the AKT gene had been significantly upregulated in the OSCC tissue samples when compared to normal tissues (p<0.05). Moreover, upregulated AKT is postulated to be involved in increased cell proliferation and reduced apoptosis in OSCC. The gene expression analysis was done in the samples of histologically confirmed OSCC, and it revealed that the AKT gene was significantly upregulated in OSCC tissues. Thus, AKT could be postulated as a potential therapeutic target for OSCC.

Abstract 1727: Silencing of novel m6A reader PRRC2A as a therapeutic strategy for oral cancer

Abstract Background: Oral squamous cell carcinoma (OSCC) poses a substantial global health challenge characterized by limited treatment options and unfavorable prognoses. Recent research has illuminated the pivotal role of N6-methyladenosine (m6A) modification and its regulators in cancer biology. However, the precise involvement of a novel m6A regulator (m6A reader), PRCC2A in OSCC remains inadequately understood. Objective: This study aims to unravel the role of PRCC2A in OSCC and ascertain its potential as both a diagnostic biomarker and a therapeutic target. Methods: We collected 76 OSCC tumor samples and 38 adjacent non-tumor samples from OSCC patients. Comprehensive analyses, including RT-qPCR, western blotting, histopathological examination, and associations with clinicopathological features, were performed to assess PRRC2A expression in cancer tissues. Furthermore, we explored its links with key factors like metastasis and its implications for patient prognosis. In vitro experiments were conducted using OSCC cell lines and human normal oral keratinocytes (NOKs). PRCC2A knockdown in OSCC cell lines was assessed for its effects on cell proliferation, apoptosis, cell cycle, migration, invasion, and sphere-forming abilities. The study also delved into its impact on immune cell infiltration and involvement in biological pathways. Results: Our investigation unveiled significant overexpression of PRCC2A in OSCC tissue samples and cell lines, relative to adjacent non-tumor tissues and NOKs (P &amp;lt; 0.001). These findings were further validated by the Cancer Genome Atlas - Head and Neck Squamous Cell Carcinoma (TCGA-HNSC) dataset. PRCC2A expression displayed significant correlations with nodal metastasis, higher tumor grades, and advanced cancer stages in OSCC samples, suggesting its crucial role in driving cancer progression and aggressiveness. Silencing PRCC2A in vitro led to reduced metastasis, diminished tumor colony formation, and increased apoptosis in oral cancer cell lines. Furthermore, the study uncovered PRCC2A intricate connections with immune regulatory genes, highlighting its potential to influence the tumor immune microenvironment. Conclusion: PRCC2A emerges as a promising candidate for both diagnosis and therapeutic intervention in OSCC. Its pronounced overexpression in oral cancer tissues, coupled with its associations with metastasis and intricate interactions with immune regulatory networks, underscores its significance in the context of oral cancer malignancies. This research offers valuable insights into the multifaceted role of PRCC2A in OSCC and lays the groundwork for future investigations and potential clinical applications. Citation Format: Balachander Kannan, Vijayashree Priyadharsini Jayaseelan, Senthil MuruganM, Paramasivam Arumugam. Silencing of novel m6A reader PRRC2A as a therapeutic strategy for oral cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1727.

Investigating the Association Between Fusobacterium nucleatum and Oral Squamous Cell Carcinoma: A Pilot Case-Control Study on Tissue Samples.

Background Fusobacterium nucleatum (F. nucleatum)has been increasingly linked to oral squamous cell carcinoma (OSCC), prompting this study to explore its presence using polymerase chain reaction (PCR) and evaluate its clinical significance. Methods In this pilot case-control study, 12 OSCC tissue samples and 12 non-cancerous oral mucosal tissue samples were analyzed. Total RNA extraction and complementary DNA (cDNA) synthesis were performed using Trizol-based methods, followed by PCR amplification and gel electrophoresis. The clinical characteristics of participants and PCR results were recorded. Results Among the OSCC tissue samples, three out of 12 tested positive forF. nucleatum,while none of the control samples showed its presence. The detection rate ofF. nucleatumin OSCC was 25%. Gel analysis confirmed specific amplicon amplification, and ImageJ software enabled copy number quantification. Discussion Our findings support previous research indicating a potential association betweenF. nucleatumand OSCC. Understanding the etiological significance ofF. nucleatumin OSCC has clinical implications, including early detection, risk stratification, and prognostication. However, the limited sample size and the need for further research to elucidate underlying mechanisms are acknowledged. Conclusion This pilot study provides initial evidence ofF. nucleatum'spresence in a subset of OSCC samples, supporting its potential association with oral cancer. DetectingF. nucleatumin OSCC tissues holds promise for future research and clinical applications as a diagnostic and prognostic biomarker. Understanding its role in oral carcinogenesis will facilitate the development of targeted therapeutic strategies. Larger studies are warranted to validate these findings and investigate the precise mechanisms involved.

Circular RNA hsa-circ-0006203 - hsa-circ-0004872 as novel detecting biomarkers in oral cancer.

Oral squamous cell carcinoma (OSCC) is the most common cancer of the head and neck region. The circular RNA (circRNA) is known to serve an important role in the carcinogenesis of different types of cancer. However, the circRNA role of OSCC remains unclear. OSCC tissues and adjacent normal tissues were obtained to detect circRNAs expression by the next generation sequencing (NGS), and OSCC tissues were selected to verify the differentially significant circRNAs by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). To further investigate the role of hsa-circ-0006203 - hsa-circ-0004872, the primer design and RT-PCR were performed. The expression levels were detected by RT-qPCR. The NGS results demonstrated that circRNAs were abundantly expressed in OSCC, and two circRNAs were significantly differentially expressed. hsa-circ-0006203 - hsa-circ-0004872 were significantly downregulated in OSCC tissue samples and was statistically correlated with pathological differentiation. In summary, the results of the present study revealed that OSCC tissues have abundant circRNAs and, to the best of our knowledge, it was our team who firstly explore the regulatory role of the hsa-circ-0006203 - hsa-circ-0004872 network in OSCC. The results indicated that hsa-circ-0006203 - hsa-circ-0004872 may be a potential biomarker for OSCC.

Oral Squamous Cell Carcinoma-Derived Cell-Free DNA Modulates Stemness and Migration of Oral Squamous Cell Carcinoma Cell Line by Inducing M2 Macrophage Polarization

To investigate the effect of oral squamous cell carcinoma (OSCC)-derived cell-free DNA (cfDNA) on the polarization of macrophages and the regulatory effect of polarized macrophages on the stemness and migration of OSCC cells. A total of 30 OSCC tissue samples, 10 dysplastic oral tissue samples, and 10 normal oral tissue samples were collected. The status of all tissue samples was confirmed by pathology analysis. Immunohistochemical (IHC) staining and immunofluorescence (IF) staining were performed to examine the cell count and location of M2 macrophages in different types of oral tissue samples. The conditioned medium (CM) of OSCC cell line CAL-27 from the human tongue was collected and the cfDNA was concentrated and isolated for identification. The macrophages were treated by cfDNA and their morphological characteristics were observed under microscope. The expression levels of polarization-related indicators were determined by RT-qPCR. CAL-27 cell line was treated with macrophage CM induced by cfDNA and the expression levels of stemness-related genes were determined by RT-qPCR. Scratch-wound assay was conducted to verify that the migration ability of CAL-27 was modulated by macrophages induced by cfDNA. There were more M2 macrophages in the deep connective tissue of dysplastic oral epithelium and the stroma of OSCC compared with those in the normal oral tissues ( P<0.05). OSCC cell line CAL-27 could secret cfDNA of 10000-15000 bp in length. cfDNA secreted by CAL-27 could induced in macrophages significantly higher expression of M2-macrophage-related genes ( P<0.05). cfDNA-treated macrophages induced significantly increased expression of stemness-related genes in CAL-27 cell line ( P<0.05) and promoted the migration ability of CAL-27 cell line ( P<0.05). OSCC-derived cfDNA promotes stemness and migration of OSCC cell line by inducing M2 macrophage polarization.

DJ-1 Oncogene as a Potential Diagnostic and Prognostic Biomarker for Head and Neck Cancer.

Background Current methods used to diagnose and prognosticate oropharyngeal cancer have contributed to unfavorable patientsurvival rates that have not significantly improved for the last several decades. Precision medicine oncology relies on molecular diagnostics and biomarkers to supplement existing methods of detecting and prognosticating cancers. This study evaluated the expression of DJ-1, an oncogene that is implicated in the pathogenesis of oral squamous cell carcinoma (OSCC), the most common type of head and neck cancer, to determine its utility as a diagnostic and prognostic biomarker. Methodology Immunohistochemistry (IHC) was performed on 13 normal oral mucosa tissue samples and 143 OSCC tissue samples of varying histopathological grades. Computer-assisted image analysis was performed using the Aperio ImageScope software from Leica Biosystems (Buffalo Grove, IL), which utilizes an algorithm of positive pixel counting for the quantification of immunoreactivity and the percentage of positive cell staining, generating a histo-score (H-score). The comparisons of the average H-scores of the different groups were made using a two-tailed T-test with P ≤ 0.05 set as the level of significance. Results The study found a significant increase in DJ-1 expression in oral squamous cell carcinoma tissue samples in comparison to the normal oral mucosa tissue samples. Additionally, the study documented a significant upregulation in DJ-1 expression in the OSCC tissue samples with high histopathological grades compared to the OSCC tissue samples with low histopathological grades. Conclusions DJ-1 expression patterns were able to reliably differentiate between oral squamous cell carcinoma and the normal counterpart tissues of the oral mucosa, thereby highlighting its role as a potential diagnostic biomarker. Moreover, DJ-1 expression significantly correlates with the OSCC histological grade, which serves as an indicator of the differentiation status and a predictor of the biological behavior of malignant neoplasms, adding to DJ-1's potential utility as a prognostic biomarker for this common type of head and neck cancer.

CCNB2 as a potential biomarker of bladder cancer via the high throughput technology.

Bladder cancer and oral squamous cell carcinoma (OSCC) seriously affect people's health. However, the relationship between bladder cancer and OSCC remains unclear. Got GSE138206, GSE146483, GSE184616, and bladder cancer datasets GSE65635, GSE100926 from Gene Expression Omnibus database. Weighted gene co-expression network analysis was used to identify the significant module. Functional enrichment analysis was performed via the Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes. Furthermore, the Gene Set Enrichment Analysis was also used to complete the enrichment analysis. Comparative Toxicogenomics Database found most relevant diseases to core genes. TargetScan is used to forecast analysis of microRNA and target genes. In Gene Ontology analysis, differentially expressed genes were mostly concentrated in cell differentiation, extrallular region, structural molecule activity, and actin binding. In Kyoto Encyclopedia of Genes and Genomes analysis, the differentially expressed genes were mainly enriched in PI3K-Akt signaling pathway, pathway in cancer, and extracellular matrix-receptor interaction. Seven hub genes (cyclin B2 [CCNB2], TK1, CDC20, PCNA, CKS1B, CDCA5, MCM4) were obtained. Hub genes (CCNB2, CDC20) are highly expressed in OSCC and bladder cancer samples. CCNB2 was one common oncogene of bladder cancer and OSCC.

Pathogenetic model of survivin-dependent molecular signalling pathways in tumorigenesis of oral cancer and precursor lesions.

p53 tumour suppressor gene limits unchecked cellular growth in response to DNA damage, by causing G1 arrest and the activation of apoptosis. Inhibitors of apoptosis include survivin which acts by inhibition of caspases. Survivin has a significant role as a cell cycle modulator and is only minimally present in mature tissues. Aberrant expression of p53 and survivin has been evaluated in various carcinomas. Thus, the objective of this research was to elucidate the co-expression of p53 and survivin in tissue samples of Oral Potentially Malignant Disorders (OPMDs) and Oral Squamous Cell Carcinoma (OSCCs). Thirty tissue samples of OPMDs and 30 tissue samples of OSCCs taken from department archives were used in the study. Expression of p53 and survivin was analyzed in the study groups by the help of immunohistochemistry. Also, co-expression of both the markers was evaluated. The expression of p53 and survivin in the oral epithelium of patients with OSCCs was significantly higher than that in patients with OPMDs (P value ≤0.05). Our results provide insights into the altered survivin and p53 co-expression with significant immunoexpression within the study groups. Therefore, survivin and p53 could be better markers for identifying cell proliferation and apoptotic pathway. Also, malignant transformation rate of OPMD increases with increased expression of these markers.

Prognostic Value of WNT1, NOTCH1, PDGFRβ, and CXCR4 in Oral Squamous Cell Carcinoma.

The biological behaviour of oral squamous cell carcinoma (OSCC) might be an effect of aberrant activation of several signalling pathways, like WNT and NOTCH pathways. The purpose of this study was to investigate the prognostic significance of WNT1 and NOTCH1 in patients with OSCC in relation to pro-angiogenic factors (PDGFRβ, CXCR4). Immunohistochemistry was performed in 60 OSCC tissue samples to compare WNT1, NOTCH1, PDGFRβ, and CXCR4 expression and correlate it with clinicopathological parameters. Immunoreactivity of WNT1, NOTCH1, PDGFRβ, and CXCR4 was found in 51.7%, 25.0%, 63.3%, and 70.0% of the patients, respectively. WNT1 expression was positively correlated with NOTCH1 (r=0.269, p=0.037) and CXCR4 (r=0.268, p=0.038). Positive correlation was observed also between NOTCH1 and CXCR4 (r=0.493, p<0.001). WNT1 and PDGFRβ expression was significantly associated with shorter overall survival (p=0.042, p=0.033, respectively). Multivariate Cox regression model revealed that WNT1 and CXCR4 were independent prognostic factors (p=0.009, p=0.021, respectively). WNT1 and NOTCH1 expression is associated with CXCR4 expression in OSCC, suggesting that WNT and NOTCH pathways are important in oral tumour angiogenesis. Moreover, WNT1 and CXCR4 could serve as independent prognostic factors for patients with OSCC.

Inverse correlation of CD3 & vascular endothelial growth factor in incisional oral squamous cell carcinoma biopsies predicts nodal metastasis & poor survival of patients.

Oral squamous cell carcinoma (OSCC) is widely prevalent in the Indian subcontinent mainly due to habit-associated aetiologies. Immune regulation and angiogenesis are the part of tumourigenesis that play a crucial role in metastasis and survival. However, the concurrent expression of vascular endothelial growth factor (VEGF) and CD3 (immune regulator receptor on T-lymphocyte) in the same OSCC tissue samples has not been reported in the Indian population. The present study evaluated the expression of CD3+ T-cells and VEGF in OSCC tissue samples and studied the clinicopathological correlation and survival analysis in an Indian population. This was a retrospective study conducted on 30 formalin-fixed and paraffin embedded sections which were histologically diagnosed as OSCC cases comprising of 15 metastatic OSCC and 15 non-metastatic OSCC with available clinical data and survival status. Reduced expression of CD3+ T-cells and increased VEGF expression were observed in metastatic OSCC samples. The correlation of expression of CD3+ T-cells and VEGF with clinicopathological parameters showed a significant association between these markers with age, nodal status, site of the lesion and survival. Reduced expression of CD3+ T-cells in OSCC was found to be associated with a significantly poor survival. VEGF was found to be over expressed in metastatic OSCC as compared to that in non-metastatic OSCC. The study findings suggest that the evaluation of CD3 and VEGF in incisional OSCC biopsies can be considered for predicting the survival outcome and metastasis.

Evaluation of Superoxide Dismutase, an Antioxidant Enzyme, in Oral Squamous Cell Carcinoma: A Systematic Review and Meta-Analysis

Objective: The objective is to qualitatively and quantitatively review the published literature on superoxide dismutase (SOD) levels in oral squamous cell carcinoma (OSCC). Materials and Methods: MEDLINE (via PubMed), SCOPUS, Science Direct, Cochrane Library, ProQuest, Google Scholar, and gray literature were comprehensively searched. Human clinical studies published between 2000 and 2021 were included. The Newcastle–Ottawa Scale was used to evaluate the risk of bias in the studies. Results: Of the 873 articles screened, six studies were included, evaluating 640 patients (554 OSCC cases and 113 controls). SOD levels in OSCC subjects were significantly lesser (P &lt; 0.01) compared to healthy controls in spectrophotometry analysis, whereas its immuno-expression was significantly higher in the OSCC tissue samples in immunohistochemistry. In the meta-analyses related to SOD analysis by spectrophotometry, the overall effect size of tissue SOD in OSCC and control group were 8.51 and 11.62 U/mg protein, respectively. In the meta-analyses related to SOD analysis by immunohistochemistry, the overall effect size of tissue SOD in OSCC was 55.55% and 3.61% for cell staining, respectively. Conclusion: Decreased SOD levels in OSCC tissue samples in spectrophotometry and high immunoexpression in immunohistochemistry accurately reflect increased oxidative stress (OS) levels. Further studies are required to understand the increase in SOD levels in OSCC tissue. Correlation analysis between SOD levels and established clinicopathological prognostic markers could aid in formulation of OS-based prognostication and treatment planning.

Exosome-mediated long noncoding RNA (lncRNA) PART1 suppresses malignant progression of oral squamous cell carcinoma via miR-17-5p/SOCS6 axis.

Exosomes derived from oral squamous cell carcinoma (OSCC) could modulate OSCC development. This study aimed to explore effects of exosome-mediated lncRNA PART1 on OSCC cells. This study was performed in Tianjin Medical University Cancer Institute from February 2021 to March 2022. Bioinformatic analysis was performed on the public database GEPIA (http://gepia.cancer-pku.cn/). Exosomes isolated from cell lines squamous cell carcinoma 9 (SCC9) and Centre Antoine Lacassagne-27 (CAL27) were identified by transmission electron microscope and western blot. Exosome-mediated lncRNA PART1, microRNA-17-5p(miR-17-5p) and suppressor of cytokine signaling 6(SOCS6) RNA expressions were assessed by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Cell counting kit8(CCK-8), caspase-3 activity, and flow cytometry were applied to evaluate OSCC cell viabilities and apoptosis. Meanwhile, OSCC cell migratory ability and invasiveness were evaluated using transwell assay. Bindings between miR-17-5p and lncRNA PART1 or SOCS6 were validated using the luciferase reporter test. Western blot was used for detecting the protein levels of SOCS6, phosphorylated signal transducer and activator of transcription 3 (STAT3) and STAT3. : According to GEPIA, lncRNA PART1 was downregulated in OSCC tissue samples and cells, and it had a positive correlation with the good prognosis of Head and neck squamous cell cancer (HNSCC) patients. After the exosomes from OSCC cells were isolated and verified, PART1 was then confirmed to be secreted by exosomes. Further, overexpression of exosome-mediated lncRNA PART1 inhibited OSCC cell viabilities, migration, and invasiveness but facilitated OSCC cell apoptosis. PART1 upregulated SOCS6 through sponging miR-17-5p. Moreover, exosome-mediated lncRNA PART1 suppressed the phosphorylation of STAT3. Exosome-mediated lncRNA PART1 could mediate the OSCC progression via miR-17-5p/SOCS6 axis in vitro, suggesting that lncRNA PART1 might be a target for treating OSCC.

Long non-coding RNA XIST promotes the malignant features of oral squamous cell carcinoma (OSCC) cells through regulating miR-133a-5p/VEGFB.

Oral squamous cell carcinoma (OSCC) represents a frequently seen oral cavity malignancy, and the mechanisms of its occurrence and development remain unclear. The present work examined the expression and biological function of long non-coding RNA (lncNRA) XIST (X-inactive specific transcript) in OSCC cells and tissues. A total number of 50 OSCC and paired non-carcinoma tissue samples were collected in this study. Gene expression levels in cancer tissues and cells were quantified by RT-qPCR. In addition, gain- and loss-of-function experiments were conducted to investigate the biological roles of XIST as well as its downstream targets in OSCC cells. XIST was upregulated in OSCC cells and tissues, which predicted a poorer prognostic outcome in OSCC patients. Silencing XIST inhibited the growth and invasion of OSCC cells and triggered apoptosis. miR-133a-5p was identified as a downstream target of XIST, which was downregulated in OSCC tissues. miR-133a-5p mediated the effect of XIST by targeting VEGFB. VEGFB overexpression rescued the inhibitory effects of XIST silencing on cell growth, invasion and migration. Taken together, the above data indicates that XIST serves as an oncogenic factor to enhance the growth and invasion of OSCC cells by targeting the miR-133a/VEGFB axis.

Ekspresija survivina u oralnom skvamocelularnom karcinomu

Introduction: Survivin functions as an apoptosis inhibitor and a regulator of cell division. This study aimed to determine the correlation between survivin expression and clinicopathologic parameters of oral squamous cell carcinoma (OSCC) and determine its potential role in the progression/prognosis of this type of tumor. Materials and methods: Immunohistochemical analysis of survivin expression was performed on 45 surgically obtained paraffin-embedded tissue samples of OSCCs. Data on patients' gender, age, tumor grade, site and stage, disease recurrence, metastasis occurrence , and disease-free interval (DFI) were correlated to survivin expression. Results: Survivin immunoreactivity was observed in 77.8% of samples. No significant correlation between survivin expression and age (p = 0.087), gender (p = 0.334), tumor site (p = 0.175), presence of lymph node metastases (p = 0.201), or disease recurrence (p = 0.451) was found. Survivin expression was observed in well and moderately differentiated tumors and in all clinical stages (p = 0.139). Patients with low survivin expression had better survival rates than the group with medium and high survivin expression, i.e., there was a tendency of a shorter DFI in patients with higher expression of survivin (p = 0.065). Conclusion: There is a tendency for a shorter disease-free period in patients with higher survivin expression. These data suggest that survivin expression in OSCC may act as an additional prognostic parameter that indicates an increased proliferative tumor potential. To further validate survivin as a prognostic marker in OSCC, a study with a larger sample size along with clinical follow-up data is needed.

Localization of Fusobacterium nucleatum in oral squamous cell carcinoma and its possible directly interacting protein molecules: A case series.

While 15 to 20% of cancers are associated with microbial infection, the relationship between oral microorganisms and oral squamous cell carcinoma (OSCC) remains unclear. The location of bacteria in a tumor is closely related to its carcinogenic mechanism. The aim of this study was to analyse bacterial diversity in clinical OSCC tissue samples and tumor distant normal tissues, locate target bacteria, and search for proteins that may interact with target bacteria. The 16S rDNA method was used to analyse bacterial diversity in clinical OSCC tissue samples and tumor distant normal tissues. Correlations between Fusobacterium abundance and clinicopathological characteristics were analysed using the χ2 test. The position of target bacteria was analysed by fluorescence in situ hybridization (FISH), and the expression of CK, CD31, CD45, CD68, cyclin D1, β-catenin, E-cadherin, NF-κB, and HIF-1α was analysed by immunohistochemistry (IHC) in OSCC tumor tissues and tumor distant normal tissues. The 16S rDNA results showed that the detected amount of Fusobacterium in OSCC tumor tissues was significantly larger than that in tumor distant normal tissues. High expression of Fusobacterium was significantly correlated with the lifestyle-related oral risk habits, including smoking (p=0.036) and alcohol consumption (p=0.022), but did not correlate with patient sex, age, tumor laterality, tumor size, grade or TNM stage. Fusobacterium nucleatum was enriched in tumor stroma, where CD31+ blood vessels and inflammatory cells (including CD45+ leukocytes and CD68+ macrophages) were densely distributed. Cyclin D1 was mainly expressed in the nucleus of tumor cells. β-catenin was expressed in the tumor cell membrane and was positively expressed in tumor interstitial vascular endothelial cells. E-cadherin was mainly expressed in tumor cell membranes. NF-κB was positively expressed in the cytoplasm of tumor cells, tumor interstitial cells and myo-fibrocytes. HIF-1α was mainly expressed in the cytoplasm of tumor interstitial cells. HIF-1α was highly expressed where Fusobacterium nucleatum was densely distributed. According to our study, the detected amount of Fusobacterium in OSCC tumor tissues was significantly larger than that in tumor distant normal tissues, and Fusobacterium nucleatum might aggravate inflammation and hypoxia by interacting with NF-κB and HIF-1α in OSCC.

Gene network analysis of the transcriptome impact of methylated microRNAs on oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is one of the most common head and neck squamous cell tumors. MicroRNAs and DNA methylation, as epigenetic mechanisms, regulate the expression of oncogenes and tumor suppressor genes, contributing to the carcinogenic development. However, the current knowledge on the genetic and epigenetic landscape of OSCC is still limited. To assess the transcriptomic impact of microRNAs found to be methylated through Infinium genome-wide methylation profiling of archived OSCC tissues, and to analyze their biological role using gene network analysis. We used the Infinium array-based methylation assay to assess the genome-wide methylation status at the single-CpG-site level of DNA purified from archived OSCC tissue samples. After quality control, filtering out poorly performing probes and normalization of data, we identified the differentially methylated microRNA loci. We performed a literature-based analysis of OSCC transcriptomic data to identify the predicted target genes for each microRNA, followed by individual network and pathway enrichment analyses. The analysis of Infinium methylation array data revealed 1469 differentially hypomethylated loci, 4 of which were of interest, namely hsa-microRNA-124-3, hsa-microRNA-24-1, hsa-microRNA-769, and hsa-microRNA-4500. Network and pathway enrichment analyses revealed multiple pathways modulated through DNA methylation-microRNA expression axes. We describe the transcriptomic impact of 4 differentially methylated microRNAs in OSCC tissues samples and discuss their role in the pathology of OSCC. These results may contribute to a better understanding of how epigenetic mechanisms such as DNA methylation and microRNAs cooperate to impact the development of OSCC.