Oral Squamous Cell Carcinoma Group Research Articles

Oral Microbial Changes in Oral Squamous Cell Carcinoma: Focus on Treponema denticola, Lactobacillus casei, and Candida albicans

Background and Objectives: In this study, we aimed to explore the oral bacteria and fungi that can help discern oral squamous cell carcinoma (OSCC) and investigate the correlations between multiple key pathogens. Materials and Methods: Twelve participants (8 females and 4 males; mean age, 54.33 ± 20.65 years) were prospectively recruited into three groups: Group 1: healthy control, Group 2: patients with stomatitis, and Group 3: patients with OSCC, with 4 individuals in each group. Unstimulated whole saliva samples from these participants were analyzed using real-time PCR to assess the presence and abundance of 14 major oral bacterial species and Candida albicans. Results: The analysis revealed significant differences for certain microorganisms, namely, Treponema denticola (T. denticola), Lactobacillus casei (L. casei), and Candida albicans. T. denticola was most abundant in the OSCC group (5,358,692.95 ± 3,540,767.33), compared to the stomatitis (123,355.54 ± 197,490.86) and healthy control (9999.21 ± 11,998.40) groups. L. casei was undetectable in the healthy control group but was significantly more abundant in the stomatitis group (1653.94 ± 2981.98) and even higher in the OSCC group (21,336.95 ± 9258.79) (p = 0.001). A similar trend was observed for C. albicans, with DNA copy numbers rising from the healthy control (464.29 ± 716.76) to the stomatitis (1861.30 ± 1206.15) to the OSCC group (9347.98 ± 5128.54) (p = 0.006). The amount of T. denticola was positively correlated with L. casei (r = 0.890, p < 0.001) and C. albicans (r = 0.724, p = 0.008). L. casei’s DNA copy number was strongly correlated with C. albicans (r = 0.931, p < 0.001). These three oral microbes exhibited strong positive correlations with each other and had various direct or indirect relationships with other species. Conclusions: In the OSCC group, T. denticola, L. casei, and C. albicans exhibited strong positive correlations with one another, further emphasizing the need for a deeper understanding of the complex microbial interactions in the OSCC environment.

Association of serum and salivary dipeptidyl peptidase-4 (DPP-4) with oral cancerous and precancerous lesions; an observational diagnostic accuracy study

BackgroundEnhancing the prognosis and treatment outcomes of oral cancer relies heavily on its early detection. This study aims to establish a connection between serum and salivary dipeptidyl peptidase-4 (DPP-4) levels and oral squamous cell carcinoma (OSCC), comparing them with oral potentially malignant lesions (OPMLs) and control subjects and validating salivary DPP-4 as a diagnostic marker for the early detection of oral cancer.MethodologyForty-five systemically healthy individuals were categorized into three groups: Group I consisted of 15 patients diagnosed with OSCC, Group II comprised 15 patients with OPMLs (including leukoplakia and oral lichen planus), and Group III included 15 participants without any oral mucosal lesions. Serum and whole unstimulated salivary samples were collected from all participants to assess DPP-4 levels using an enzyme-linked immunosorbent assay (ELISA) kit. Receiver operating characteristic (ROC) analysis was conducted to determine the area under the curve (AUC), sensitivity, specificity, and diagnostic accuracy of DPP-4.ResultsBoth serum and salivary DPP-4 levels were highest in the healthy group, followed by OPMLs, and lowest in the OSCC group, with statistically significant differences observed. ROC analysis demonstrated excellent diagnostic accuracy of salivary DPP-4 in distinguishing OSCC from healthy individuals, OPMLs from healthy individuals, and OSCC from OPMLs, with accuracies of 100%, 100%, and 96.67%, respectively. Salivary DPP-4 levels also exhibited a statistically significant correlation with OSCC grades.ConclusionsDPP-4 appears to play a protective, anti-oncogenic role in maintaining oral tissue health. The remarkable diagnostic accuracy of both serum and salivary DPP-4 in discriminating between OSCC, OPMLs, and healthy controls suggests its potential utility as a well-established marker for early oral cancer diagnosis. Salivary DPP-4, being non-invasive, could serve as a convenient chair-side diagnostic tool for the early detection of OSCC.Clinical trial registrationThe study was retrospectively registered on clinicaltrial.gov with NCT06087042, date of registration (first posted date): 17/10/2023

Immunohistochemical Expression of Bcl-2, E-cadherin, CD34 and CD105 in Basaloid Squamous Cell Carcinoma - An In Vitro Study

Abstract Introduction: Basaloid squamous cell carcinoma (BSCC) is a rare aggressive variant of oral squamous cell carcinoma (OSCC) with a high propensity for distant metastasis. In this article, we present clinicopathological and survival data of eight cases of BSCC and further analyse the behaviour of these tumours with the help of E-cadherin, CD34, CD105 and B cell lymphoma-2 (Bcl-2) immunoexpression. Materials and Methods: Histopathologically confirmed cases of BSCC were retrieved from the department archives. Clinicopathological details and survival data of these patients were collected. Immunohistochemical analysis was performed with Bcl-2, E-cadherin, CD34 and CD105 on these cases and compared with different grades of OSCC (well differentiated, moderately differentiated and poorly differentiated). The statistical analysis was done using IBM SPSS software version 23. Results: BSCC was seen commonly in males of age group 49–71 years and predominantly reported in the retromolar trigone. Bcl-2 expression was significantly lower in BSCCs when compared to the conventional OSCC groups (P < 0.05). E-cadherin expression showed no significant difference between BSCC and well-differentiated OSCC group (P = 0.487). The overall mean survival for patients with BSCC was 6.37 months. Discussion: BSCCs of the oral cavity show increased CD105, CD34, E-cadherin and low Bcl-2 labelling. A substantial relationship between the tumour neo-angiogenesis, collective cell migration and apoptotic property could be related to the aggressive nature of this tumour and its poor overall survival rate. BSCCs are common in middle to older aged male and show increased expression of CD105, CD34 and E-cadherin.

Role of Antioxidant Enzymes in Pathogenesis of Oral Squamous Cell Carcinoma: a Systematic Review and Meta-analysis.

To investigate the antioxidant enzyme status in biological samples of patients with oral squamous cell carcinoma (OSCC) and compare them with biological samples of healthy people through a systematic review and meta-analysis. Antioxidant enzymes of catalase (CAT), sodium dismutase (SOD) and glutathione peroxide (GPx) were included in the analysis. A literature search was conducted of the PubMed, Science Direct, Scopus, Web of Science and Wiley Online Library databases for studies published between January 1999 and December 2022. A total of 831 articles were selected, of which 131 were found to be relevant. Finally, the full texts of 12 studies were screened and included. Studies that evaluated other antioxidant enzymes were excluded. Standardised mean difference (SMD) was derived to conduct a meta-analysis using comprehensive meta-analysis v3 (Biostat, Englewood, NJ, USA). A random effects model with 95% confidence interval (CI) was used to estimate the effect size. P < 0.05 was considered significant. CAT levels were measured in eight studies (n &#61; 567) and the mean values for the OSCC and control groups were 4.81 ± 2.57 and 10.02 ± 1.81, respectively (SMD 3.18, 95% CI 1.01 to 1.42; P &#61; 0.001). SOD level was evaluated in 11 studies (n &#61; 762) and the values for the OSCC and control groups were 3.78 ± 1.45 and 7.34 ± 1.79, respectively (SMD 3.66, 95% CI 1.51 to 1.94; P &#61; 0.001). GPx level was evaluated in 10 studies (n &#61; 697) and the values for the OSCC and control groups were 13.33 ± 1.42 and 16.54 ± 2.9, respectively (SMD 1.91, 95% CI 1.34 to 1.77; P &#61; 0.001). The heterogeneity between the studies was severe (I2 ≥ 90%). The risk of bias between studies was low to moderate. Analysis revealed that the levels of antioxidant enzymes decreased in biological samples of patients with OSSC as compared to healthy controls. Understanding the pathological progress of OSCC by analysing the level of antioxidant enzymes is beneficial in formulating a personalised, targeted pro-oxidant therapy for cancer treatment.

Diagnostic value of 8-OHdG, 8-iso-PGF2α and TNF-α levels in blood for early carcinogenesis of erosive oral lichen planus.

Oral cancer has a high worldwide incidence and mortality rate showing an upward trend year by year, predominantly occurring in emerging countries. Oral squamous cell carcinoma (OSCC) is one of the main types of oral cancer, accounting for more than 90% of all cases in oral cancer. To evaluate the diagnostic value of 8-hydroxy-2'-deoxyguanosine (8-OHdG), 8-iso-Prostaglandin F2alpha (8-iso-PGF2α) and tumor necrosis factor (TNF)-α as biomarkers in the early carcinogenesis of erosive oral lichen planus (EOLP) by measuring their levels in the blood of patients with EOLP and oral squamous cell carcinoma (OSCC). A total of 69 patients were enrolled in this case-control study [including an OSCC group (n= 23), an EOLP group (n= 23), and an age- and gender-matched healthy control group (n= 23)]. Blood levels of 8-OHdG, 8-iso-PGF2α and TNF-α were measured using enzyme-linked immunosorbent assay (ELISA). Statistical differences in these indicators among the three groups were analyzed. Plasma levels of 8-OHdG and 8-iso-PGF2α in the OSCC group were significantly higher than those in both the EOLP group and the control group (all P< 0.05); no significant statistical difference was found between the EOLP group and the control group. Serum levels of TNF-α in both the OSCC and EOLP groups were elevated compared with the control group, showing significant differences among all three groups (all P< 0.05). Receiver operating characteristic (ROC) curves revealed that plasma 8-OHdG and 8-iso-PGF2α levels and serum TNF-α levels had diagnostic effects on early carcinogenesis in EOLP patients. When these indicators were combined for diagnosis, the diagnostic effect was enhanced, with an area under the ROC curve (AUC) values of 0.819. 8-OHdG, 8-iso-PGF2α and TNF-α may serve as biological indicators for monitoring the early carcinogenesis of EOLP, and the diagnostic effect was augmented when these indicators were combined.

Role of Cytokeratin 19 Fragment Antigen 21-1 (CYFRA 21-1) in Early Diagnosis of Oral Squamous Cell Carcinoma

Abstract Objective: Oral cancer is considered one of the most common and rising cancers in India. Cytokeratin (CK) 21-1 (CYFRA 21-1) fragments are known to be a potential biomarker for the detection and diagnosis of potentially malignant lesion disorders (PMLDs) and oral squamous cell carcinoma (OSCC). In this study, we estimated CYFRA 21-1 levels in tobacco abusers/smokers, PMLD patients, and OSCC patients and compared with the samples of healthy controls to estimate their role in early diagnosis of OSCC. Materials and Methods: Serum CYFRA 21-1 was done by sandwich ELISA method. The kits were purchased from Everon Life Sciences. Results: Serum CYFRA 21-1 levels were seen to be raised in the PMLD and OSCC groups when compared with the healthy controls. Conclusion: Serum CYFRA 21-1 levels were raised in PML and OSCC group when compared with the healthy control. CYFRA 21-1 can be used as an essential biomarker for an early diagnosis of OSCC.

Evaluation of salivary biomarker interleukin-6 in oral squamous cell carcinoma and oral potentially malignant disorders − A comparative cross-sectional South Indian study

Abstract Background: Oral squamous cell carcinoma (OSCC) accounts for nearly 90% of oral malignancies and represents a major global health care problem. It is often preceded by oral potentially malignant disorders (OPMD). Although regular clinical examination forms the backbone for oral cancer screening, subtle lesions go unnoticed and there is a need for more sensitive and specific molecular biomarkers in mass screening of population. Salivary proteomics offer an attractive alternative to serum and tissue testing. Aims: To find the diagnostic utility of salivary interleukin-6 (IL-6) in the differential diagnosis of OSCC, OPMD from healthy controls. Study Design: In vivo study. Methods: After approval from the Institutional Review Board, unstimulated whole saliva was collected from 90 subjects, 30 in each group of OSCC, OPMD and controls after ethical clearance. Salivary IL-6 was measured by ELISA, and the results were statistically analysed. Results: Significant difference in salivary IL-6 was seen between OSCC, OPMD and controls. Receiver operating characteristic curve analysis showed the highest area under a curve of 0.982 in distinguishing OSCC from controls. It showed a sensitivity of 71% and specificity of 100% at a cut-off value of 33.4 pg/mL (P = 0.000). Moderately differentiated OSCC (MDSCC) showed a significant increase in salivary IL-6 concentration compared to well-differentiated OSCC (WDSCC). Conclusion: Results of the present study showed strong predictive power of salivary IL-6 in distinguishing OSCC from controls. Its levels also increased with tumor aggressiveness from WDSCC to MDSCC. Thus, salivary IL-6 could have a diagnostic and/or prognostic significance in identifying high-risk groups in mass screening of the population.

Identification of Cuproptosis-related Gene Lipoyltransferase 1 as a Promising Biomarker in Oral Squamous Cell Carcinoma.

To find efficient cuproptosis-related biomarkers to explore the oncogenesis and progression of oral squamous cell carcinoma (OSCC). All the original data were downloaded from the Cancer Genome Atlas (TCGA) database. Univariate Cox analysis and Kaplan-Meier survival analysis were used to identify the gene related to survival. Tumor Immune Estimation Resource 2.0 (TIMER 2.0) was used to reveal the different expression of cuproptosis-related gene lipoyltransferase 1 (LIPT1) in various kinds of tumours. LIPT1, as a cuproptosis-related gene, was found to be differentially expressed in the OSCC group and the control group. It was also found to be related to the prognosis of OSCC. Pan cancer analysis showed LIPT1 was also involved in various kinds of tumours. All the results demonstrate that the cuproptosis-related gene LIPT1 is highly involved in the oncogenesis and progression of OSCC. These findings give new insight for further research into the cuproptosis-related biomarkers in OSCC.

Analysis of Oral and Gut Microbiome Composition and Its Impact in Patients with Oral Squamous Cell Carcinoma.

The impact of gut and oral microbiota on the clinical outcomes of patients with oral squamous cell carcinoma (OSCC) is unknown. We compared the bacterial composition of dental plaque and feces between patients with OSCC and healthy controls (HCs). Fecal and dental plaque samples were collected from 7 HCs and 18 patients with OSCC before treatment initiation. Terminal restriction fragment-length polymorphism analysis of 16S rRNA genes was performed. Differences in bacterial diversity between the HC and OSCC groups were examined. We compared the occupancy of each bacterial species in samples taken from patients with OSCC and HCs and analyzed the correlation between PD-L1 expression in the tumor specimens and the occupancy of each bacterial species. The gut and oral microbiota of patients with OSCC were more varied than those of HCs. Porphyromonas and Prevotella were significantly more abundant in patients with OSCC than in HCs. The abundance of Clostridium subcluster XIVa in the gut microbiota of the PD-L1-positive group was significantly greater than that in the PD-L1-negative group. The oral and gut microbiomes of patients with OSCC were in a state of dysbiosis. Our results suggest the possibility of new cancer therapies targeting these disease-specific microbiomes using probiotics and synbiotics.

Quantitative Assessment of Receptors of Advanced Glycation End Products Expression in Tissue Samples from Patients with oral Submucous Fibrosis, Leukoplakia, and Oral Squamous Cell Carcinoma.

Oxidative stress markers have been firmly established as elevated in oral squamous cell carcinomas (OSCCs). These markers play a crucial role in the pathogenic mechanism underlying the accumulation of advanced glycation end products (AGEs) and their respective receptors. The primary objective of this study is to discern and compare the levels of receptors of AGEs (RAGEs) within tissue samples from patients diagnosed with oral submucous fibrosis (OSMF) at varying stages, oral leukoplakia at various stages, and OSCC. A cross-sectional investigation was conducted, enrolling a total of 49 patients, distributed across three distinct groups. Tissue samples were meticulously collected from the aforementioned patient groups. Subsequently, these samples underwent a process of homogenization and centrifugation. The supernatant obtained was subjected to enzyme-linked immunosorbent assay analysis to precisely determine the concentration of RAGE. The concentration of RAGEs was found to be significantly higher at various stages of OSMF when compared to the reference group of OSCC (P < 0.05). This difference was statistically significant, indicating a substantial association. In contrast, the levels of RAGE in patients with hyperkeratosis accompanied by epithelial dysplasia at various stages were observed to be lower than those in the OSCC group, with the difference in concentration being statistically insignificant (P > 0.05). This comprehensive study has provided compelling evidence demonstrating the heightened levels of RAGE in OSMF when compared to OSCC. These findings collectively suggest the potential utility of anti-RAGE interventions as a promising avenue for novel therapeutic strategies in potentially malignant disorders such as OSMF.

Abstract 705: miR-21-5p induces radioresistance through reinforced DNA damage response in oral squamous cell carcinoma

Abstract Cancers arising from oral cavity and oropharynx are commonly treated with radiotherapy, whereas tumor radioresistance results in poor outcome. The prerequisite for developing more effective interventions for these patients is that the molecular mechanisms leading to a radioresistant phenotype in these tumors are clear to researchers. Unfortunately, the radioresistant mechanisms remain elusive to date. We recently identified that a highly conserved and cancer-related microRNA, i.e. miR-21-5p, plays an important role in radioresistance of oral squamous cell carcinoma (OSCC). We analyzed the RNA-sequencing data of 290 tissue samples from TCGA database, and found the expression of miR-21-5p was significantly upregulated in many OSCC samples (N=271) when comparing to adjacent normal tissues (N=19). The difference was further confirmed on our freshly collected OSCC tissues from patients in operation room. We performed RNA-sequencing analysis in 11 OSCC samples and their adjacent normal tissues. Among the 2106 differentially expressed genes between OSCC and normal control groups, we found that several important DNA repair genes (RAD50 double strand break repair protein gene, RAD50; DNA Polymerase Theta, POLQ; Ataxia Telangiectasia and Rad3-related, ATR) were significantly overexpressed in OSCC samples (P&amp;lt;0.05), while positive correlations between miR-21-5p expression and DNA repair genes RAD51 and XRCC4 were identified in the OSCC cohort. Notably, ATR is an important DNA damage response gene, while XRCC4, RAD51, and POLQ are involved in DNA double strand break (DSB) repair through non-homologous end joining, homologous recombination, and microhomology-mediated end joining, respectively. OSCC cell lines were further recruited to study the relations of miR-21-5p with radiation-induced DSB repair. By analyzing kinetics of 53BP1 foci, a biomarker for detecting DSBs, we found that anti-miR-21-5p transfected cells showed more residual foci, indicating of unfinished repair of DSBs, at 5 hours and 18 hours post irradiation. The deficient DSB repair resulted from anti-miR-21-5p transfection were further confirmed by single cell gel electrophoresis with comet assay. Consistently, the same treatment resulted in fewer clonogenic survivors in colony formation assay, demonstrating increased radiosensitivity of OSCC cells when miR-21-5p was antagonized. This work provided novel insights about the mechanism of miR-21-5p-induced radioresistance in OSCC, which may give rise to novel therapeutic strategy for improved treatment of OSCC patients. Citation Format: Qi Liu, Weitao Hu, Xinghan Li, Yongqiang Deng, Lin Ma. miR-21-5p induces radioresistance through reinforced DNA damage response in oral squamous cell carcinoma [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 705.

Association between Vitamin D Receptor Polymorphism and Susceptibility to Oral Lichen Planus and Oral Squamous Cell Carcinoma.

Oral squamous cell carcinomas (OSCC) comprise 90-95% of oral cancers. Early diagnosis improved the survival rate of OSCC patients to 80-90%. Oral lichen planus (OLP) is a chorionic inflammatory disease with malignancy potential. The vitamin D receptor (VDR) plays a critical role in the pathogenesis of oral cancer. This study aimed to determine the association between VDR rs7975232 (Apa I) polymorphism and potential susceptibility to OLP and OSCC risks. In this prospective case-control study, a total of 120 blood samples were obtained from OSCC patients (n=29), OLP (n=50), and controls (n=40). VDR rs7975232 polymorphism was studied using the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) method. Statistical analysis was performed with SPSS Version 23 software. Data were expressed as means ± standard deviation (SD). Age, sex, allelic frequency, and genotyping were compared using the chi-square test. A p-value of less than 0.05 was regarded as statistically significant. The disease risk was estimated by Odds ratio (OR) with a 95% confidence interval. A significant age difference was observed between the controls and the OSCC group (p=0.001). A significant difference was observed in Aa and aa genotypes compared with AA between OSCCs and controls. Moreover, dominant (p<0.001), additive (p<0.001), and allelic (p=0.001) models were different between groups. There was a positive association between rs7975232 VDR polymorphism and susceptibility to OSCC. More experimental evidence must reveal the possible association between rs7975232 and the risk of OLP in a larger cohort.

Evaluation of Mir-1290 as a Possible Diagnostic Factor in the Serum of Oral Squamous Cell Carcinoma Patients with Qualitative Real-Time Polymerase Chain Reaction.

This study aimed to determine the incidence of microRNA (miRNA; miR-1290) in the serum of oral squamous cell carcinoma (OSCC) patients compared to a control group using the qualitative real-time polymerase chain reaction (PCR) method. Blood serum samples were obtained from patients diagnosed with OSCC and confirmed through biopsy. The samples were collected from patients referred to the Mashhad Dental Faculty and Ghaem Hospital. The OSCC group consisted of 17 patients, while the healthy group included 15 individuals. RNA was extracted from the patient samples, and samples with an A260/280 ratio between 1.8 and 2.0 (indicating acceptable RNA quality) were immediately converted into complementary DNA (cDNA) using albumin and cDNA reference genes. The SYBR green real-time reverse transcriptase PCR method was used to measure the presence of miR-1290 in the blood samples. A total of 32 patients were examined in this study, including 17 women (53.1%) and 15 men (46.9%). The mean age was 46.7 years in the healthy group and 54.6 years in the SCC group, indicating a significant difference (P<0.05). The expression level of the miR-1290 gene was higher in patients with SCC compared to the healthy group (P=0.000). While the expression level of miR-1290 was higher in grade 3 and advanced stage than in grades 2 and 1 and early stage, the differences were not statistically significant (P=0.173 and P=0.564 for grade and stage, respectively). The expression level of miR-1290 may increase in SCC patients compared to healthy individuals, making it a potential circulating biomarker. Further investigations for diagnostic utility would be warranted.

Correlation between human papillomavirus protein expression and clinicopathological features in oral squamous cell carcinoma.

Given the implications of concurrent human papilloma viral infection (HPV) in the prognostic course and implications on therapeutic approached of patients with oral squamous cell carcinoma (OSCC), we seek to investigate the implications that P16 expression has on the clinical course and pathological appearance of patients with OSCC and concurrent infection. Using S-P immunohistochemistry, we examined the expression of P16 and Ki67 in 460 patients with OSCC. We compared the expression of the protein between the tumor cells and normal epithelial mucosa within the same patient. The clinical and pathological characteristics (including gender, age, histological grade, lymph node metastasis, clinical stage, clinical recurrence, tumor diameter, Ki67 proliferation index) were analyzed by stratification statistically. In total 460 cases of OSCC were identified and expression of P16 was significantly higher in the OSCC group compared to the normal mucosal epithelial group (X2 = 60.545, p = .000). There also appear to be a gender predilection as the expression was higher in females compared to males (0.218 vs. 0.144, X2 = 3.921, p = .048). Younger age also appears to be a predictive factor as those under 35 years old had higher expression of the protein compared to those over 35 years old (0.294 vs. 0.157, X2 = 4.230, p = .040). P16 positivity showed a significant positive correlation with histologic grade (X2 = 4.114, p = .043). In addition, the positive rate of P16 was higher in patients with ki67 over 85% (0.455 vs. 0.160, X2 = 6.667, p = .023). OSCC with HPV infection tends to occur more frequently in female patients and those under 35 years of age. HPV infection with expression of the P16 and ki67 protein may promote the proliferation and growth of OSCC at a higher frequency.

Gene mutation analysis of oral submucous fibrosis cancerization in Hainan Island.

The sequencing panel composed of 61 target genes was used to explore the related mutation genes of oral squamous cell carcinoma (OSCC) and oral submucous fibrosis (OSF) cancerization, so as to provide a theoretical basis for the early diagnosis of oral submucous fibrosis cancerization, find the most important mutations in OSF cancerization, and more targeted prevention of OSF cancerization. A total of 74 clinically diagnosed samples were included, including 36 cases of OSCC and 38 cases of OSF cancer patients. DNA was extracted, and targeted gene panel sequencing technology was used to analyze the gene frequency of pathogenic mutation sites in clinical samples. Gene panel sequencing analysis showed that there were 69 mutations in 18 genes in OSCC and OSF cancerous specimens. The results of gene panel sequencing were screened, and 18 mutant genes were finally screened out and their mutation frequencies in the samples were analyzed. According to the frequency of gene mutations from high to low, they were TP53, FLT4, PIK3CA, CDKN2A, FGFR4, HRAS, BRCA1, PTPN11, NF1, KMT2A, RB1, PTEN, MSH2, MLH1, KMT2D, FLCN, BRCA2, APC. The mutation frequency of FLT4 gene was significantly higher than that of OSCC group (P < 0.05). FLT4 gene may be related to OSF cancerization and is expected to be an early diagnostic biomarker for OSF cancerization.

Oral Squamous Cell Carcinoma Lipid Evaluation in Gingiva Tissue Stored in TRIzol via Shotgun Lipidomics and MALDI Mass Spectrometry Imaging.

Oral squamous cell carcinoma (OSCC) is the prevalent type of oral cavity cancer, requiring precise, accurate, and affordable diagnosis to identify the disease in early stages, Comprehending the differences in lipid profiles between healthy and cancerous tissues encompasses great relevance in identifying biomarker candidates and enhancing the odds of successful cancer treatment. Therefore, the present study evaluates the analytical performance of simultaneous mRNA and lipid extraction in gingiva tissue from healthy patients and patients diagnosed with OSCC preserved in TRIzol reagent. The data was analyzed by partial least-squares discriminant analysis (PLS-DA) and confirmed via matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). The lipid extraction in TRIzol solution was linear in a range from 330 to 2000 ng mL-1, r2 > 0.99, intra and interday precision and accuracy <15%, and absolute recovery values ranging from 90 to 110%. The most important lipids for tumor classification were evaluated by MALDI-MSI, revealing that the lipids responsible for distinguishing the OSCC group are more prevalent in the cancerous tissue in contrast to the healthy group. The results exhibit the possibilities to do transcriptomic and lipidomic analyses in the same sample and point out important candidates related to the presence of OSCC.

Assessment of salivary levels of ErbB2 in oral squamous cell carcinoma.

Oral cancer is the sixth-most common cancer globally. The survival rate of oral cancer is 5 years, depending on the stage it is diagnosed. To diagnose in the early stage, specialised tumour markers may assist and also help in improving the survival rate of oral cancer. ErbB2 is a transmembrane cell surface receptor required in signal transduction and an essential part of signalling pathways that take part in controlling the basic cellular processes like cell cycle, migration, metabolism and survival, besides cellular proliferation and differentiation. It is over-expressed in oral squamous cell carcinoma (OSCC) and is directly proportional to the poor prognosis, as it is expressed at a very low concentration in a healthy individual. Due to this, ErbB2 could be used as a diagnostic marker in OSCC. Nowadays, the search for tumour expression in the saliva with the use of salivary biomarkers could aid in the diagnosis of the OSSC. To assess the expression of ErbB2 in the saliva of patients with oral squamous cell carcinoma by correlating the ErbB2 level in the disease group with the healthy group. To determine the diagnostic significance of ErbB2 in OSCC. The study comprises 20 salivary samples from OSCC patients and 20 salivary samples from healthy individuals. The salivary level of ErbB2 was estimated using Enzyme Linked Immunosorbent Assay. To analyse the data, SPSS (IBM SPSS Statistics for Windows, Version 26.0, Armonk, NY: IBM Corp. Released 2019) is used. The significance level is fixed at 5% (α = 0.05). P value <0.05 is considered to be statistically significant. To compare the mean values of mean and concentration, an unpaired/independent sample t-test was used. The mean age of OSCC and control were found to be 57 ± 8.13 and 26.6 ± 1.51, respectively. The mean age was compared between OSCC and control by the Chi-square test, and the P value was <0.01, which was found to be statistically significant. The salivary levels of ErbB2 in the OSCC and control groups were measured by an unpaired sample t-test. The mean salivary ErbB2 level in the OSCC group is 3.20 ng/ml ± 0.57, and in the control group, it is 2.43 ng/ml ± 0.13. When a pairwise comparison of ErbB2 concentration was performed between OSCC and control, it showed a statistically significant difference with a P value of 0.007, which is P < 0.05. The present study has demonstrated an increased salivary expression of ErbB2 in OSCC patients when compared to healthy individuals. This suggests that ErbB2 could aid in the diagnosis of OSCC and could be used as a diagnostic marker in the early detection of oral cancer, a finding that has to be further established with a larger sample size.

Serum autoantibody profiling of oral squamous cell carcinoma patients reveals NUBP2 as a potential diagnostic marker.

Oral Squamous Cell Carcinoma (OSCC), a common malignancy of the head and neck region, is frequently diagnosed at advanced stages, necessitating the development of efficient diagnostic methods. Profiling autoantibodies generated against tumor-associated antigens have lately demonstrated a promising role in diagnosis, predicting disease course, and response to therapeutics and relapse. In the current study, we, for the first time, aimed to identify and evaluate the diagnostic value of autoantibodies in serum samples of patients with OSCC using autoantibody profiling by an immunome protein array. The utility of anti-NUBP2 antibody and tissue positivity in OSCC was further evaluated. We identified a total of 53 autoantibodies with significant differential levels between OSCC and control groups, including 25 that were increased in OSCC and 28 that were decreased. These included autoantibodies against Thymidine kinase 1 (TK1), nucleotide-binding protein 2 (NUBP2), and protein pyrroline-5-carboxylate reductase 1 (PYCR1), among others. Immunohistochemical validation indicated positive staining of NUBP2 in a large majority of cases (72%). Further, analysis of OSCC data available in TCGA revealed higher NUBP2 expression correlated with better disease-free patient survival. In conclusion, the differential serum autoantibodies identified in the current study, including those for NUBP2, could be used as potential biomarkers for early diagnosis or as screening biomarkers for OSCC pending investigation in a larger cohort.

The association of epidermal growth factor variant with oral squamous cell carcinoma.

In this study, our aim was to investigate the epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) gene polymorphisms in oral squamous cell carcinoma (OSCC) patients and non-OSCC healthy controls. This case-control study comprised 89 OSCC and 107 healthy controls by using polymerase chain reaction (PCR) and restriction fragment length polymorphism methods, the genotypes for EGF + 61 A > G (rs4444903) and EGFR R497K (rs2227983) were analyzed. According to the EGF + 61 A > G genotype distribution, individuals with the GG genotype were more prevalent in the OSCC group when compared to the healthy controls. But the AA genotype frequency was significantly higher in the healthy control group. The frequency of G allele carriers was 2.3 times higher than A allele carriers in OSCC patients (p < .001). For the EGFR R497K genotype, there was no significant difference between the OSCC and healthy control groups. Regarding the study results, the G allele of EGF + 61 A > G polymorphism was associated with OSCC. Larger populations and functional investigations should be used to explore the nature of the interaction between EGF and OSCC.

Expression of the Embryonic Cancer Stem Cells' Biomarkers SOX2 and OCT3/4 in Oral Leukoplakias and Squamous Cell Carcinomas: A Preliminary Study.

Cancer stem cells (CSCs) are incriminated for initiating the process of carcinogenesis either de novo or through the transformation of oral potentially malignant disorders (OPMDs) to oral squamous cell carcinoma (OSCC). The aim of this study was to detect the expression of embryonic-type CSC markers OCT3/4 and SOX2 in OSCCs and oral leukoplakias (OLs), the most common of OPMDs. The study type is experimental, and the study design is characterized as semiquantitative research, which belongs to the branch of experimental research. The experiment was conducted in the Department of Oral Medicine/Pathology, School of Dentistry, Aristotle University of Thessaloniki, Greece. This study focuses on the semiquantitative immunohistochemical (IHC) pattern of expression of CSCs protein-biomarkers SOX2 and OCT3/4, in paraffin embedded samples of 21 OSCCs of different grades of differentiation and 30 cases of OLs with different grades of dysplasia, compared to five cases of normal oral mucosa in both terms of cells' stain positivity and intensity. Statistical analysis wasperformed through SPSS 2017 Pearson Chi-square and the significance level was set at 0.05 (p=0.05). The expression of the respective genes of SOX2 and OCT3/4 was studied through quantitative polymerase chain reaction (qPCR), in paraffin-embedded samples of 12 cases of OLs with mild/non dysplasia and 19 cases moderately/poorly differentiated OSCCs(n=19) and five normal mucosa using the Independent Paired T-test. The genes SOX2 and Oct3/4 were expressed in all examined cases although no statistically significant correlations among normal, OL and OSCC, were established. A nuclear/membrane staining of OCT3/4 was noticed only in three out of 21 OSCCs but in none of OLs or normal cases (without statistical significance). A characteristic nuclear staining of SOX2 was noticed in the majority of the samples, mostly in the basal and parabasal layers of the epithelium. SOX2 was significantly detected in the OSCCs group (strong positivity in 17/21) than in the OL group (30 cases, mostly mildly stained) (p-value=0.007), and the normal oral epithelium (mild stained, p=0.065). Furthermore, SOX2 was overexpressed in well differentiated OSCCs group (5/OSCCs, strongly stained) rather than in mildly dysplastic and non-dysplastic OLs samples (14/OLs, mildly stained) (p-value =0.035). The characteristic expression of SOX2 but not of OCT3/4 in OLs' and OSCCs' lesions suggests the presence of neoplastic cells with certain CSC characteristics whose implication in the early stages of oral tumorigenesiscould be further evaluated. The clinical use of SOX2, as prognostic factor, requires further experimental evaluation in larger number of samples.