Oral Squamous Cell Carcinoma Group Research Articles

Changes in oral microbial profiles associated with oral squamous cell carcinoma vs leukoplakia.

Oral squamous cell carcinoma (OSCC) is the most frequently occurring cancer among head and neck SCC worldwide. The identification of novel effective biomarkers for early detection may greatly improve the survival rate and prognosis of patients with OSCC. This study aimed to identify specific oral microbial profiles associated with OSCC. Saliva samples were collected from oral leukoplakia (OLK) and OSCC patients (N=6 each) and healthy controls (HC; N=4). Total bacterial genomic DNA was isolated and 16S rRNA gene survey was performed by next-generation sequencing of the V4 region. The relative distribution of abundance for phylogenetic groups was compared among the OSCC and OLK groups. The 448 operational taxonomic units detected from the libraries were classified into 133 genera, 69 families, 41 orders, 26 classes and 12 phyla. The abundance of phyla Bacteroidetes and genus Solobacterium was notably higher in the OSCC group when compared with the OLK group, whereas those of genus Streptococcus was significantly lower in the OSCC group when compared with the OLK. These changes in the salivary microbiome may have potential applications as a novel diagnostic tool for the early detection of OSCC.

Salivary proteomics of canine oral tumors using MALDI-TOF mass spectrometry and LC-tandem mass spectrometry.

Canine oral tumors are relatively common neoplasms in dogs. For disease monitoring and early diagnosis, salivary biomarkers are appropriate because saliva collection is non-invasive and requires no professional skills. In the era of omics, matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry (MALDI-TOF MS) coupled with liquid chromatography-tandem MS (LC-MS/MS) are suitable to identify potential disease-associated peptides and proteins. The present study aimed to use MALDI-TOF MS and LC-MS/MS to search for particular peptide mass fingerprints (PMFs) and conceivable biomarkers in saliva of dogs with early- and late-stage oral melanoma (EOM and LOM, respectively), oral squamous cell carcinoma (OSCC), benign oral tumors (BN), and periodontitis and healthy controls (CP). Pooled saliva samples in each group were used to be representative of population change. Unique PMFs were obtained and specific peptide fragments were sequenced by LC-MS/MS and BLAST-searched with mammalian protein databases. Seven peptide fragments appeared in the tumor groups (EOM, LOM, OSCC and BN) at 1096, 1208, 1322, 1794, 1864, 2354 and 2483 Da, two peptide fragments appeared in the LOM and OSCC groups at 2450 and 3492 Da, and in the CP controls at 2544 and 3026 Da. Also, protein–chemotherapy drug interaction networks were exhibited. Using western blot analysis, the expression of sentrin-specific protease 7 (SENP7), a peptide fragment at 1096 Da, in OSCC was significantly increased, as was the expression of TLR4, a peptide fragment at 3492 Da, in LOM and OSCC, compared with the CP group. The expression of nuclear factor kappa B (NF-κB), a TLR4 partner, was notably increased in OSCC compared with CP, BN and EOM. The expression was also enhanced in LOM compared with EOM. Expressed protein sequences from western blots were verified by LC-MS/MS. Western blots were then performed with individual samples in each group. The results showed the elevated expression of TLR4 in LOM and OSCC, compared with that in CP and BN, the increased expression of NF-κB in LOM and OSCC, compared with CP and in LOM compared with BN, and the enhanced expression of SENP7 in LOM and OSCC, compared with that in CP and BN. In conclusion, discrete clusters of EOM, LOM, OSCC, BN and CP groups and potential protein candidates associated with the diseases were demonstrated by salivary proteomics. Western blot analysis verified SENP7, TLR4 and NF-κB as potential salivary biomarkers of canine oral tumors.

Non-invasive screening of a microRNA-based dysregulation signature in oral cancer and oral potentially malignant disorders

IntroductionWe have previously shown that oral swirls are a robust source of microRNA protected by extracellular vesicles, potentially useful to detect oral squamous cell carcinoma (OSCC)-associated molecular aberration. ObjectivesTo study a developed dysregulation score and risk classification algorithm based upon a panel of OSCC-associated microRNA in oral swirls from individuals with OSCC and oral potentially malignant disorders (OPMDs). Materials and methodsAn OSCC-associated panel of 5 microRNAs (miR-24; miR-21; miR-99a; let-7c; miR-100;) was quantified by qPCR in 190 individuals with and without mucosal abnormalities, including OSCC (n = 53) and OPMDs (n = 74). Each sample was analyzed using a developed dysregulation score (dSCORE) and risk classification algorithm, allocating a LOW- or HIGH-RISK score. The influence of demographic, systemic, oral health and mucosal disease factors on the developed test was analyzed. ResultsMicroRNA for analysis can be predictably isolated from oral swirls sourced from individuals with a range of demographic, systemic and oral health findings. Utilizing the presence of HIGH-RISK identified OSCC patients with 86.8% sensitivity and 81.5% specificity. Older age and female gender were associated with higher dSCOREs and higher proportions of HIGH-RISK classification amongst individuals with no mucosal abnormalities. The dSCOREs for all subgroups of OPMDs were significantly different from the OSCC group. ConclusionThis is the first comparison of microRNA sourced from oral swirls from individuals with OPMDs with individuals with and without OSCC. A HIGH-RISK dysregulation signature was found to be accurate in indicating the presence of OSCC and exampled to parallel malignant transformation.

Salivary Antisense Non-Coding RNA In the INK4 Locus & Maternal Expressed Gene 3 as early biomarkers in Detection of Malignant Transformation of Oral Lichen Planus

Oral lichen planus (OLP) is a potentially malignant lesion. It is crucial to properly identify patients at risk of malignant transformation. LncRNA antisense non-coding RNA in the INK4 locus (ANRIL) has been known to advance tumor progression. Another type of lncRNAs is maternal expressed gene (MEG3) which is a tumor suppressor gene that can inhibit the advancement of malignancy. The goal of this study was to outline the relative fold expression of ANRIL and MEG3 in OLP and oral squamous cell carcinoma (OSCC). The study included 20 histopathologically diagnosed OLP patients and 10 patients with OSCC. The relative fold expression of ANRIL and MEG3 in the OLP group was compared to OSCC group using qRT-PCR. The results showed that the relative fold expression of ANRIL was significantly increase in the OSCC compared to OLP group (p<0.01). ROC analysis showed that with cut-off value of 4.1 relative fold expression of ANRIL with sensitivity 0.8, specificity 0.7 and AUC 0.85. The decrease in MEG3 was insignificant. ANRIL relative fold expression could perform an essential part in OSCC progression, and could assist in the future as a potential biomarker for malignant transformation of OLP.

Assessment of Role of Human Papilloma Virus in Patients with Oral Squamous Cell Carcinoma

Background: Head and neck malignancies include malignancies instigating in the oral cavity, the orophar-ynx, the hypopharynx and the larynx. We planned the present study to assess the role of human papilloma virus (HPV) in patients with oral squamous cell carcinoma (OSCC).&#x0D; Methods: The present study included assessment of role of human papilloma virus in patients with oral squamous cell carcinoma. A total of 50 histopathologic proven cases of well differentiated OSCC were included in the present study. A total of 50 healthy oral mucosal tissues (controls) were also included in the present study. The controls were obtained while surgical removing the impacted teeth. DNA was obtained from the tissue sections and underwent Polymerization chain reaction for identification of HPV 16/18. All the results were summarized in Microsoft excel sheet and were analyzed by SPSS software.&#x0D; Results: HPV was absent in all the cases of control group, whereas among the OSCC group, the HPV was detected in 46 percent of the cases. In the remaining 54 per-cent of the cases of the OSCC group, HPV was absent. Significant results were obtained while comparing the presence of HPV in between the OSCC group and the control group.&#x0D; Conclusions: HPV was present in sig-nificant number of OSCC cases. Therefore; HPV might play a contributing role in the etio-pathogenesis of OSCC.

Expression of programmed cell death-ligand 1 in oral squamous cell carcinoma and oral leukoplakia is associated with disease progress and CD8+ tumor-infiltrating lymphocytes

ObjectiveIn recent years, monoclonal antibodies targeting programmed cell death-ligand 1 (PD-L1) have become a promising cancer immunotherapy. However, the role of PD-L1 in oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMDs), including oral leukoplakia (OLK), remains controversial. The aim of the present study was to investigate the expression level of PD-L1 in OSCC and OPMDs, and examine its relationship with CD8 expression and different clinicopathological features. MethodExpression of PD-L1 and CD8 were conducted in 41 OSCC, 21 OLK, and 25 normal mucosa samples by immunohistochemistry. Then, the density of PD-L1 expression was measured, and its correlation with CD8 expression and different clinicopathological features was analyzed. ResultsPD-L1 protein was detected in 97.6% of OSCC, 61.9% of OLK, and 0% of normal tissues. PD-L1 was highly expressed in human OSCC tissue (P < 0.0001), when compared to both OLK and control tissues. PD-L1 positivity was significantly associated with CD8 density (P < 0.0001, r = 0.8491). The PD-L1 high expression OSCC group displayed a trend for improved overall survival (OS) and disease-free survival (DFS) compared to the low expression group, although the differences were not significant. Moreover, the expression level of PD-L1 in OSCC was positively correlated with the pathological grade (P < 0.0001), but it was independent of age, gender, smoking, drinking, tumor size, lymph node status, or recurrence (P > 0.05). Also, there was a significant upregulation of PD-L1 expression observed in the OLK group compared to the control group (P < 0.0001). PD-L1 positivity in OLK patients was associated with gender and smoking habits (P < 0.05), but it did not correlate with age, drinking, or dysplasia (P > 0.05). ConclusionThe upregulation of PD-L1 may be associated with disease progress and CD8+ tumor-infiltrating lymphocytes in oral premalignant and malignant lesions.

Evaluation of enzymatic and non-enzymatic antioxidant status and biomarkers of oxidative stress in saliva of patients with oral squamous cell carcinoma and oral leukoplakia: a pilot study

Objective: The study aimed to evaluate total antioxidant capacity as well as levels of various enzymatic and non-enzymatic antioxidants, and oxidative stress markers in saliva of patients with oral squamous cell carcinoma (OSCC) and oral leukoplakia (OL).Material and methods: Twenty patients with OSCC, 20 patients with OL and 20 healthy subjects were enrolled into this prospective study. Total Antioxidant Capacity (TAC), activity of superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR) as well as levels of total glutathione (tGSH), reduced glutathione (GSH), oxidized glutathione (GSSG), GSH/GSSG ratio, uric acid (UA), 8-hydroxy-2′-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) were estimated in saliva using appropriate biochemical methods.Results: The activity of SOD was significantly higher in OSCC group in comparison with OL and control groups. The levels of GSH were markedly lower in OSCC and OL patients as compared to the control group. Likewise, we found that GSH/GSSG ratio was markedly lower in the OSCC and OL groups. Levels of some biomarkers were influenced by clinical staging of OSCC and OL as well as by sociodemographic factors.Conclusions: The results of this pilot study suggest that salivary activity of SOD is higher in OSCC patients, whereas levels of GSH and GSH/GSSG ratio are lower in saliva of patients with OSCC and OL. Clinical staging of OSCC and OL, as well as some sociodemographic factors may also influence salivary antioxidant status.

In vivo detection of oral precancer using a fluorescence-based, in-house-fabricated device: a Mahalanobis distance-based classification.

In vivo detection of oral precancer has been carried out by a fluorescence-based, in-house-developed handheld probe on three groups: oral squamous cell carcinoma (OSCC), dysplastic (precancer), and control (normal). Measurements have been performed on a total of 141 patients and volunteers of different age groups. Excitation wavelength of 405nm was used and fluorescence emission spectra were recorded in the scan range of 450.14 to 763.41nm at very low incident power (122μW) from different oral sites buccal mucosa (BM), lateral boarder of tongue (LBT), and dorsal surface of tongue (DST). Spectral profiles are found to vary among the three groups as well as among the different oral sites. Major and minor bands of flavin adenine dinucleotide (FAD) and porphyrins near 500, 634, 676, 689, and 703nm have been obtained. Porphyrin contribution is found to be more dominant than the FAD in OSCC and dysplastic groups as compared to the control group. A better classification has been observed using the entire spectral range rather than restricting to individual bands, by application of principal component analysis (PCA), Mahalanobis distance model, and receiver operating characteristic analysis (ROC). ROC on Mahalanobis distance differentiates OSCC to normal, dysplastic to normal, and OSCC to dysplastic with sensitivities from 71% to 98%, 92% to 94% and 81% to 93% and specificities 91% to 100%, 86% to 100% and 79% to 97% for oral sites BM, LBT and DST. LBT and DST appear to be more sensitive to dysplasia detection as compared to BM.

Evaluation of candidal species among individuals with oral potentially malignant disorders and oral squamous cell carcinoma.

Context:Cancer afflicts almost all communities worldwide. Although it arises de novo in many instances, a significant proportion of oral squamous cell carcinoma (OSCC) develops from potentially malignant disorders (PMDs). Further, the association of Candida with various potentially malignant and malignant lesions has been reported as a causative agent.Aims:The aim of the study is to evaluate and intercompare the predominant candidal species among individuals with PMD and OSCC.Subjects and Methods:The swab samples were collected for the microbiological culture followed by incisional biopsy for histopathological confirmation. The swab samples were streaked and incubated on Sabouraud-dextrose agar medium and positive candidal colonies were incubated on CHROM agar for speciation.Settings and Design:A total of clinically diagnosed 95 subjects of which 25 as normal controls, 30 as PMDs and 40 as OSCC were included. The collected swab samples were initially streaked and incubated on Sabouraud dextrose agar (SDA) medium, and later, only positive candidal colonies were incubated on CHROM agar for speciation.Statistical Analysis:Chi-square test was utilized.Results:Positive candidal growth on SDA medium was seen in 24%, 43% and 82% and negative in 76%, 57% and 18% individuals of normal controls, PMDs and OSCC, respectively. On evaluation on Chromagar medium, Candida species was present in 20%, 40% and 77% and absent in 80%, 60% and 23% individuals among controls, PMDs and OSCC group, respectively. On speciation of Candida in CHROMagar among the controls, PMDs and OSCC, Candida albicans species was present in 4 (16%), 7 (23%) and 4 (10%); Candida krusei in 1 (4%), 5 (17%) and 10 (25%); Candida glabrata in nil, nil and 6 (20%) and Candida tropicalis in nil, nil, and 2 (5%) cases, respectively.Conclusion:There was predominant carriage of candidal species in PMDs and OSCC, but whether Candida has specific establishment in PMDs or in malignancy is still a matter of debate.

Oral Candidal Carriage Among Patients with Oral Squamous Cell Carcinoma: A Case-Control Study

Introduction: Etiology of oral squamous cell carcinoma is multifaceted. The established risk factors include tobacco, alcohol, and immunodeficiency states with infections gaining an importance. Candida species has been implicated in oral squamous cell carcinoma (OSCC). The present study aims at determining the prevalence of Candida species in the saliva of patients with OSCC and healthy cohorts. Materials and Methods: Unstimulated saliva was collected from patients with OSCC (n = 90) and age and sex matched healthy subjects (n = 170). The samples were inoculated onto sabouraud dextrose agar and incubated for a week. The isolates were enumerated using a colony counter. The isolates were identified using standard phenotypic methods. The significance of oral candidal carriage was calculated using chi-square test. Odds and risk ratio was calculated using pearson’s chi-square test. Result: Oral candidal carriage were present in 70% of patients with oscc, while healthy cohorts had a prevalence of 20%. The oral candidal carriage among OSCC patients in comparison to healthy cohorts was highly significant. (p = 0.0001). C. albicans and non-albicans Candida were equally distributed in the OSCC group. Colony-forming units were high among patients with oscc. Significant odds and risk ratio was observed for the prevalence of Candida species among OSCC (p

Tumor necrosis factor-like weak inducer of apoptosis expression in healthy oral mucosa, oral dysplasia and oral squamous cell carcinoma.

Objective:Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) has been implicated in the pathogenesis of cancer, as it participates in the progression of internal malignancies. However, its role in the biology of squamous cell carcinoma (SCC) is uncertain. Studies regarding TWEAK in SCC have shown inconsistent results. We aimed to study the expression of TWEAK in healthy oral mucosa, oral dysplastic lesions and in oral SCC (OSCC).Methods:Immunohistochemistry for TWEAK was performed on one hundred oral mucosal tissues, healthy control (HC) (n = 20), oral dysplasia (OD) (n = 20) and OSCC (n = 60). Staining intensity, extent of staining (ES) and immunoreactive Score (IRS) were assessed for each sample. Kruskal–Wallis ANOVA, Mann–Whitney U, Chi-square and Spearman's rank correlation coefficient were applied.Results:TWEAK was expressed in 55% of HC, 90% of OD and in all cases of OSCC, with variable intensities. A significant difference in the ES and IRS of TWEAK was noted among the three groups. ES and IRS were highest in OSCC group. ES of TWEAK was significantly higher at invasive tumor front (ITF) than in the whole tumor, with a significant positive correlation. TWEAK expression showed a significant association with invasive front grading, pattern of invasion and surgical margins of OSCC.Conclusions:TWEAK may contribute to the progression of OSCC. It might also sustain altered differentiation, invasion and migration of tumor cells at ITF.

Immunohistochemistry of YAP and dNp63 and survival analysis of patients bearing precancerous lesion and oral squamous cell carcinoma.

Background: Yes-associated protein (YAP) is a candidate oncogene in various human cancers, and recently, it has been reported that YAP expression and its activity was enhanced by ΔNp63. However, the role of YAP and ΔNp63 expression in carcinogenesis and progression of oral squamous cell carcinoma (OSCC) has been unknown. Therefore, we investigated how YAP and ΔNp63 influence carcinogenesis and progression of OSCC.Methods: We performed immunohistochemical analyses in whole tissue samples to investigate YAP and ΔNp63 expression in normal oral mucosa, epithelial hyperplasia, oral epithelial dysplasia (OED; low/high grade), carcinoma in situ (CIS), and OSCC. Furthermore, in OSCC, we analyzed clinical significance by using Kaplan-Meier survival analysis.Results: In normal oral mucosa and epithelial hyperplasia, YAP expression was primarily confined to the basal and parabasal layers, but YAP expression was elevated in OED, CIS, and OSCC. In OED, YAP and ΔNp63 expression levels were markedly higher in high grade than in low grade. In OSCC groups, YAP and ΔNp63 expression patterns tended to differ according to histopathological differentiation of OSCC. Furthermore, the YAP high expression group, which showed YAP staining in >50% positive cells with strong cytoplasmic staining or >10% positive cells with nuclear reactivity, showed a tendency to have a poor survival rate.Conclusion: YAP and ΔNp63 expression levels correlated with grade of oral OED. Additionally, YAP expression was associated with OSCC survival rate. Our results suggested that YAP and ΔNp63 expression might serve as predictive markers to distinguish OSCC development and progression.

Prognostic value of intratumour and intra field heterogeneity rate in predicting second events in oral squamous cell carcinoma

Event Abstract Back to Event prognostic value of intratumour and intra field heterogeneity rate in predicting second events in oral squamous cell carcinoma Andrea Gabusi1*, Davide Bartolomeo Gissi1, Roberto Rossi1 and Luca Morandi1 1 University of Bologna, Department of Biomedical and Neuro-Muscular Sciences - Section of Oral Science, Italy Aim. Improvements in sequencing technologies have revealed that genetic differences among neoplastic cells may reflect clonal expansion and thereby the coexistence of multiple subclones in a single tumour. Prognostic implications of intratumour heterogeneity are not fully understood. Aim of this project was to investigate if the degree of intratumour heterogeneity was related to tumour progression in oral squamous cell carcinoma (OSCC). Materials and Methods. Patients operated for OSCC and attending regular follow up for disease relapse at the Unit of Oral Medicine and Maxillofacial Surgery S.Orsola Hospital, University of Bologna were recruited for the study in order to include both recurrent and non recurrent OSCC. Inclusion criteria: diagnosis of primary tumor of T2-T4 according to the p-TNM classification of tumours, absence of nodal involvement, history of non-smokers, absence of lichenoid inflammation in tumour microenvironment, surgical margin of resection free from neoplasia, Follow up ≥ 3 years after surgical resection. Multiple samples were obtained from each tumour and from related adjacent non neoplastic mucosa. Next generation sequencing for mutational analysis was applied for 10 OSCC tumour specific genes: KRAS ,NRAS, HRAS, BRAF, PIK3CA, TP53, NOTCH1, PTEN , CDKN2A, EGFR. The degree of ITH for OSCC somatic mutations was derived by calculating the heterogeneity rates (HR) of all affected genes and dividing that sum by the number of affected genes not shared by all tumor regions. These values were used to compare the ITH in the group of recurrent and non recurrent OSCCs. Results. HR values between groups were then statistically compared using the non parametric U-Mann-Whitney test for independent samples (IBM® SPSS Software v.21). P value in primary tumours scored 0,095 and did not reach significativity indicating the absence of a statistical difference in the degree of intratumor heterogeneity among the groups of recurrent and non recurrent OSCC.By contrast, U-Mann-Whitney test performed on samples from non neoplastic adjacent mucosa disclosed that non recurrent OSCC tends to exhibit statistically lower values of HR if compared to recurrent OSCCs ( p= 0,032). Discussion. According to our results , in primary tumours, no particular degree of heterogeneity was able to distinguish OSCC with aggressive local behaviors. Genetic instability in tumours may produce heterogeneous subclones but not all subclones may reflect aggressive behaviors. On the other hand, ITH analysis of non neoplastic adjacent mucosa disclosed that non recurrent OSCC tends to exhibit statistically lower values of HR if compared to recurrent OSCCs ( p= 0,032). Fields of less aggressive OSCCs are thus more homogeneous. A prognostic implication of genetic heterogeneity of pre neoplastic field is here documented for the first time.

Estimation of expression of beta-human chorionic gonadotropin levels through progression of disease from normal to epithelial dysplasia to malignancy.

Background:Gonadotropins have been extensively studied in trophoblastic and nontrophoblastic tumors of breast, gastric, bladder, parathyroid, renal cell and cervical carcinomas, with a significant increase in tissue expressions. Serum levels of beta-human chorionic gonadotropin (β-hCG) and its tissue expression were found more in oral squamous cell carcinoma (OSCC) patients with a significant diagnostic and prognostic value. No such study has been done on oral epithelial dysplasia (OED).Aims and Objective:To evaluate the expression of β-hCG in OED and the feasibility of using this marker for early diagnosis and to see its progression from normal to dysplasia to malignancy.Materials and Methods:The study population consisted of thirty histologically confirmed cases of OED and thirty cases of OSCC. Fifteen normal tissues were also included in the study. All the tissue samples were subjected to immunohistochemical (IHC) staining using antimouse β-hCG antibody.Results:The IHC expression of β-hCG was completely negative in normal cases (Group 1 [n = 15]), whereas 13 (43.3%) cases of OED (Group 2 [n = 30]) and 13 cases (43.3%) of OSCC (Group 3 [n = 30]) showed diffuse cytoplasmic staining in dysplastic surface epithelium and epithelial islands of OSCC. This difference was statistically significant with P = 0.007.Conclusion:We conclude that the expression of β-hCG increased from normal mucosa to dysplasia to OSCC, suggesting that it is involved in the early stage of carcinogenesis and progression of the disease.

Vascular density and inflammatory infiltrate in primary oral squamous cell carcinoma and after allogeneic hematopoietic stem cell transplantation.

Hematopoietic stem cell transplantation (HSCT) recipients have been reported to have an increased risk of chronic graft versus host disease (cGVHD) and hematological and solid cancers. Oral manifestations are the first signs of cGVHD observed in the majority of patients, and oropharyngeal cancer is the most frequent secondary malignancy occurred after HSCT. In this study, we have evaluated the inflammatory infiltrate cell content and correlated with the vascular density in patients affected by primary oral squamous cell carcinoma (OSCC) from previous healthy controls and OSCC after cGVHD. Results have demonstrated that patients with OSCC after GVHD show a more consistent inflammatory infiltrate as compared with the OSCC ones. In detail, the inflammatory background composed of CD3-positive T cells, tryptase-positive mast cells, CD31-positive endothelial cells, and CD68-positive macrophages may be more pronounced in the setting of GVHD + OSCC than in the control group. By contrast, CD20-positive B cells and CD1a-positive dendritic cells were more abundant in the latter population. Finally, a positive correlation was found as between vascular density and inflammatory cell infiltration in both GVHD + OSCC and OSCC groups. Overall, these results confirm the role played by immune cells in enhancing tumor progression and angiogenesis and suggest a potential therapeutic strategy involving inhibition of recruitment of immune cells to the tumor microenvironment and blockade of pro-tumoral effects and pro-angiogenic functions.

Correlation of plasma lipid levels and histopathological grading in patients with oral squamous cell carcinoma and its pre-cancerous lesions

&lt;p class="abstract"&gt;&lt;strong&gt;Background:&lt;/strong&gt; Lower levels of lipid in proliferating tissues and in blood compartments may occur due to ongoing process of oncogenesis. The evidence regarding hypolipidemia as a predisposing factor for malignancy is scarce. Therefore, this study was undertaken to study alteration of plasma lipid levels in patients diagnosed with oral squamous cell carcinoma (OSCC) and its oral pre-cancerous (OPC) lesions and also to compare the plasma lipid profile in different grades of OSCC.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Methods:&lt;/strong&gt; A single centre prospective cross sectional study was carried out among 150 consecutive patients of OSCC and OPC lesions. The diagnosis of all lesions was confirmed histopathologically and grading was also done for each lesion based on degree of differentiation and keratinization of tumour cells. Serum total cholesterol, HDL, TG, LDL was estimated by using semi-automated chemical analyzer. The lipid levels were compared by using unpaired t-test between two strata whereas one way analysis of variance (ANOVA) with Tukey’s post-hoc tests was used among more than two strata. &lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Results:&lt;/strong&gt; Total cholesterol, TG, HDL, and LDL were significantly reduced in OSCC and OPC groups when compared with the reference values. Serum TC, LDL and HDL levels were found to be decreased with the loss of differentiation in histological grading but with no statistical significance.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Conclusions:&lt;/strong&gt; It can be inferred that there is an inverse relationship between the serum lipid profile values of TC, HDLC, TG, LDL and OSCC as well as OPC patients. The lower serum lipid status may be considered a useful indicator for initial changes occurring in neoplastic cells.&lt;/p&gt;

Discovery and preclinical validation of proteomic biomarkers in saliva for early detection of oral squamous cell carcinomas.

Our aim was to identify and prevalidate a set of salivary proteins that can distinguish oral squamous cell carcinomas (OSCC) patients from healthy individuals and patients with oral potentially malignant disorders (OPMD). Proteomes of 60 saliva samples from healthy individuals, OPMD patients, and OSCC patients were assayed using the isobaric tags for relative and absolute quantitation (iTRAQ) method. Enzyme-linked immunosorbent assay (ELISA) was used to prevalidate the candidate biomarkers in an independent sample set (n=90). In total, 246 differentially expressed proteins were identified by comparing each two groups, and 21 proteins were differentially expressed when OSCC was compared with both OPMD and Control. Three proteins, namely, solute carrier family 3 member 2 (SLC3A2), S100 calcium-binding protein A2 (S100A2), and interleukin-1 receptor antagonist protein (IL1RN), were selected as candidate biomarkers. Comparing the OSCC group with the healthy group, the area under curve (AUC) of the three combined biomarkers was 0.89, with a sensitivity of 83.33% and a specificity of 83.33%. Comparing the OSCC group with the OPMD group, the AUC value was 0.87, with a sensitivity of 93.33% and a specificity of 70.00%. Our study indicates that salivary proteomics is promising for the discovery of OSCC biomarkers.

CD8+ and CD163+ infiltrating cells and PD-L1 immunoexpression in oral leukoplakia and oral carcinoma.

Overexpression of inhibitory checkpoint PD1/PD-L1 plays an important role in carcinogenesis and patients prognosis. 70 cases of oral squamous cell carcinoma (OSCC), 23 cases of oral leukoplakia (OLK), and 19 control cases were immunohistochemically stained with anti-PD-L1, -CD8, and -CD163 antibodies. PD-L1 was expressed on dysplastic and subepithelial infiltrating cells of OLK as well as on cancer and tumor-infiltrating cells of OSCC. In OSCC, PD-L1 immunoexpression was significantly increased in comparison to OLK, and control groups. The correlative study showed significant correlations between the immunoexpression of PD-L1 and the number of CD8+, CD163+ cells in both OLK and OSCC groups. We found also significant negative correlation between the number of PD-L1+ infiltrating cells and the number of CD8+ cells in OSCC, and positive correlation between the number of PD-L1+ infiltrating cells and CD163+ cells in OLK and OSCC groups. In conclusion, our study indicate that CD163+ and CD8+ infiltrating cells influence the early and subsequent stages of oral carcinogenesis. We demonstrated also that studied tumors may evade the host immune system by PD-L1 immunoexpression not only on epithelial cells but on infiltrating cells as well.

Circulating catecholamines are associated with biobehavioral factors and anxiety symptoms in head and neck cancer patients.

Studies have shown that stress-related catecholamines may affect cancer progression. However, little is known about catecholamine secretion profiles in head and neck cancer patients. The present study investigated plasma norepinephrine and epinephrine levels in head and neck squamous cell carcinoma (HNSCC) patients and patients with oral leukoplakia, as well as their association with clinicopathological and biobehavioral variables and anxiety symptoms. A total of 93 patients with HNSCC and 32 patients with oral leukoplakia were included. Plasma norepinephrine and epinephrine levels were measured by high performance liquid chromatography with electrochemical detection (HPLC-ED), and psychological anxiety levels were measured by the Beck Anxiety Inventory (BAI). Plasma norepinephrine and epinephrine concentrations were significantly higher in patients with oral and oropharyngeal squamous cell carcinoma (SCC) compared to non-cancer patients. Oral SCC patients displayed plasma norepinephrine levels about six times higher than oropharyngeal SCC patients, and nine times higher than oral leukoplakia patients (p < .001). Plasma epinephrine levels in oral SCC patients were higher compared to the oropharyngeal SCC (p = .0097) and leukoplakia (p < .0001) patients. Oropharyngeal SCC patients had higher plasma norepinephrine (p = .0382) and epinephrine levels (p = .045) than patients with oral leukoplakia. Multiple regression analyses showed that a history of high alcohol consumption was predictive for reduced plasma norepinephrine levels in the oral SCC group (p < .001). Anxiety symptom of “hand tremor” measured by the BAI was an independent predictor for higher plasma norepinephrine levels in HNSCC patients (β = 157.5, p = .0377), while the “heart pounding/racing” symptom was independently associated with higher plasma epinephrine levels in the oropharyngeal SCC group (β = 15.8, p = .0441). In oral leukoplakia patients, sleep deprivation and worse sleep quality were independent predictors for higher plasma norepinephrine levels, while severe tobacco consumption and higher anxiety levels were factors for higher plasma epinephrine levels. These findings suggest that head and neck cancer patients display sympathetic nervous system hyperactivity, and that changes in circulating catecholamines may be associated with alcohol consumption, as well as withdrawal-related anxiety symptoms.

Human papilloma virus 16/18: Fabricator of trouble in oral squamous cell carcinoma

AimTo find out the association between Human Papilloma Virus (HPV) genotypes 16/18 in Pakistani patients with oral squamous cell carcinoma (OSCC). MethodsDNA from oral rinse of 300 subjects was taken. The subjects included 100 cases with OSCC and 200 controls. Samples were analyzed by both conventional and real time PCR using “HPV consensus Gp5+/Gp6+ and HPV 16, 18 specific primers”. ResultsOut of 300 persons, 74/300 (25%) were found to be infected with HPV: “46/100(46%) from cases and 74/200(14%) from controls”. The distribution was: HPV16, 6/300 (8%): 4/100 (9%) from OSCC group and 2/200 (8%) from controls while HPV 18 was 9/300(12%): 5/100(11%) from cases and 4/200(16%) from controls. Out of 300 subjects, 26(35%) were infected by “both HPV 16/18 (23(50%) from cases and 3(12%) from controls”. Persons who were infected with HPV 16&18 had higher chances to develop OSCC as compared to those who didn’t have HPV 16/18 (AOR: 21.4, 95% CI: 5.73 – 80.8). ConclusionThe exposure to high risk strains of Human papilloma virus (16/18) in combination can be fabricotor of trouble (p<0.001, Adjusted odds ratio; 21.42) in OSCC.