Oral Squamous Cell Carcinoma Group Research Articles

HPV-16 Detection and Quantitation in Whole Mouth Fluid of Oral and Cervical Cancer Patients

Introduction: The association of human papillomavirus (HPV) in cervical and oropharyngeal cancer is well established. There is a growing evidence that HPV could possibly contribute to oral cancer along with alcohol and tobacco use. The aim of this study was to evaluate and quantify HPV-16 in the whole mouth fluid of patients with oral squamous cell carcinoma (OSCC), cervical cancer, and normal controls. Materials and Methods: The whole mouth fluid of 20 patients with OSCC (Group I), 10 with cervical cancer (Group II), and 10 normal individuals (Group III) was collected by the spit technique. The DNA was extracted and quantified by quantitative polymerase chain reaction. Data were analyzed by SPSS 21 (IBM Corp. Released 2012, Armonk, NY: IBM Corp) software and analysis of variance test was used. Results: The mean age of Group I patients was 50.9 ± 13.5 years, Group II was 51.6 ± 8.07 years, and Group III was 46 ± 8.5 years. In Group I, 15 were males and 5 were females. In Group III, 5 were males and 5 were females. Eight patients belonging to Group I, four of Group II, and seven of Group III had HPV-16 in their whole mouth fluid. Conclusion: HPV-16 was identified in the whole mouth fluid of OSCC, patients with cervical cancer, and even in normal controls. However, the mean viral loads were highest in patients with cervical cancer followed by patients with OSCC and normal controls.

Alteration in Serum Lipid Profile Pattern in Oral Squamous Cell Carcinoma and Potentially Malignant Disorders

Objective: To evaluate and compare lipid profile level in oral submucous fibrosis (OSMF), oral leukoplakia (OL) and oral squamous cell carcinoma (OSCC) patients . Material and Methods: Thirty histopathologically diagnosed subjects each of OL, OSMF, OSCC were recruited along with 30 healthy controls. 5ml of venous blood is collected and estimated using standard diagnostic kits. Results: The mean of Total cholesterol level in controls was 219.03 mg%, in OSCC, OL and OSMF was 142.89 ± 10.21mg%, 155.44 ± 17.63 mg% and 180.60 ± 13.25 mg%, respectively. The mean low-density lipid level in controls was 137.24 mg and in OSCC, OL and OSMF groups were 109.28 ± 2.16 mg%, 126.63 ± 0.85 mg% and 119.15 ± 0.93 mg%, respectively. The mean of high-density lipid level in controls, OSCC, OL and OSMF was 42.87 ± 0.42 mg%, 36.50 ± 2.31 mg%, 21.13 ± 0.77 mg% and 28.37 ± 1.11mg%, respectively. The mean of very low density lipids level in controls, OSCC, OL and OSMF was 30.12 ± 1.51 mg%, 17.24 ± 0.80 mg%, 22.25 ± 0.93 mg% and 25.89 ± 0.43 mg%, respectively. The mean triglyceride level in controls, OSCC, OL and OSMF was 118.80 ± 9.47 mg%, 91.2 ± 3.03 mg%, 105.05 ± 2.96 mg% and 106.19 ± 3.09 mg%, respectively . Conclusion: Lipid profile levels could be early indicators of precancer and cancer.

Identification and validation of potential novel biomarkers for oral squamous cell carcinoma

ABSTRACT Our study aimed to explore potential new diagnostic biomarkers in patients with oral squamous cell carcinoma (OSCC) to find new target molecules involved in the progression of OSCC. Potential novel biomarkers of OSCC were identified using a protein microarray assay. Compared with the healthy control group, there were five proteins (I309, GDF15, AXL, MMP3, and CTACK) in the serum of in situ oral cancer group. However, there were four differentially expressed proteins (MCSF, I309, MMP3, and CTACK) in the serum of the OSCC group. Receiver operating characteristic (ROC) curve analysis results suggested that these six proteins (I309, GDF15, AXL, MMP3, CTACK, and MCSF) had diagnostic value for OSCC. Based on The Cancer Genome Atlas (TCGA) database, we found that only GDF15 expression was associated with the prognosis of OSCC. Subsequently, we verified the expression levels of six proteins in HSC-3 and HaCaT cells, and the results showed that the level of these six proteins was significantly higher in HSC-3 cells than in normal HaCaT cells. Similarly, in the OSCC nude mouse model, the expression levels of these proteins were significantly upregulated in OSCC tumor tissue compared to the normal tissue. GDF15, MMP3, AXL, MCSF, I309, and CTACK may be used as biomarkers for OSCC diagnosis and provide a novel study direction for the treatment of OSCC.

Age And Gender Predilection Of Habits And Oral Cancer Among An Outpatient Population Visiting A Dental Hospital

Oral Squamous Cell Carcinoma (OSCC) is the most common neoplasm representing more than 80% of all oral cancer cases. Squamous cell carcinoma is defined as a “malignant epithelial neoplasm exhibiting squamous differentiation as characterized by the formation of keratin and/ or the presence of intercellular bridges”.Most of the epidemiological studies have revealed that heavy smoking and alcohol intake are the most important risk factors for oral cancer. Poor dental hygiene, accompanied by tooth loss, certain oncogenic viruses have been associated with an increased risk of cancer. The purpose of this study was to analyze the gender and age predilection of habits and oral cancer among an outpatient population visiting a dental college. This was a retrospective study (2019-2020) in which data on 34 Oral Squamous Cell Carcinoma(OSCC) were investigated. The patient records were reviewed and analysed between January 2019 and March 2020. All the 34 patient data were tabulated in excel, reviewed and was classified according to the border's grading system. Following which data was imported to SPSS software by IBM for descriptive statistical analysis and chi-square test was performed. A significant association between OSCC and age group (45-50 Years) was observed though it was statistically insignificant. (p value=0.64). Males have a higher incidence of OSCC compared to females in the same age group. A significant association is observed between Gutka, Smoking and pan chewing with OSCC. OSCC is the most commonly occurring oral cancer and there is a high predilection of the OSCC with age, gender and habits. Dentists play a key role in screening, detection and treatment of oral malignancies and can help in educating the public to bring about more awareness towards prevention of Oral Cancer.

Mel-CAM Expression in Common Oral Carcinomas.

Statement of the Problem: Mel-CAM (CD146, MUC18) is a 113-kD heterophilic cell-cell adhesion glycoprotein found in normal and tumoral tissues. The biologic functions and role of the Mel-CAM can be employed as a diagnostic marker in pathology.Purpose: The aim of this study was assessing the expression of Mel-CAM in common oral carcinomas like salivary gland mucoepidermoid carcinoma (MEC) and oral squamous cell carcinoma (OSCC) to differentiate the OSCC from high-grade MEC.Materials and Method: This study was performed on 19 specimens of MEC and 17 specimens of OSCC, which were retrieved from the archive of Department of Pathology of Taleghani Hospital, Tehran, Iran. Immunohistochemical staining was performed by using antibody against CD146. The data were analyzed using SPSS software through Mann-Whitney, Spearman's correlation coefficient, and Kruskal-Wallis tests.Results: Mel-CAM was expressed in all MEC samples and 10 OSCC cases. The two groups were significantly different regarding the CD146 expression (p= 0.035). Furthermore, the CD146 expression was found to be significantly correlated with the invasion mode (p= 0.002), tumor size (p= 0.012), and histological grade (p= 0.024) in OSCC group. No significant correlation existed between the expression, intensity, and location with the histological grade of MEC (p> 0.05) nor was any significant correlation detected between the CD146 expression and lymph node metastasis in neither group.Conclusion: Regarding the significant correlation between the CD146 expression and the prognostic factors in OSCC, this marker may predict the prognosis in OSCC patients, but not the MEC lesions. It cannot be used for differentiating high-grade MEC and OSCC.

Upregulated histone deacetylase 2 gene correlates with the progression of oral squamous cell carcinoma.

Histone deacetylases (HDACs) are considered as an essential regulator of cellular proliferation, differentiation, and apoptosis. The HDAC2 enzyme of Class I HDACs plays an important role in tumor progression of human malignancies. The aim of the present study was to analyze the HDAC2 gene expression in pre-oral cancer and oral squamous cell carcinoma (OSCC), and its association with clinico-pathological features. The HDAC2 protein expression was analyzed through the immunohistochemistry and western blot techniques in 82 oral pre-malignant, 90 OSCC, and 16 normal control tissues. qRT-PCR was used to quantify the mRNA fold change in all groups. The HDAC2 protein and mRNA levels were significantly higher in OSCC and pre-oral cancer groups compared to the controls. Immunostaining of HDAC2 protein was enhanced in 84.4% of OSCC and 67.1% of pre-cancerous tissue sections (p< 0.01). The mean protein level was analyzed as 1.96 ± 0.44 in oral carcinoma, 1.61 ± 0.39 in pre-cancer and 0.96 ± 0.10 in control tissues. In addition, HDAC2 mean protein level was associated with histological differentiation (OR = 25, p< 0.05) and tumor-node-metastasis (TNM) stages (OR = 6.2, p< 0.05) of OSCC patients. The upregulated HDAC2 gene in pre-cancer and OSCC tissues indicates its crucial role in the transformation of pre-malignant to malignant carcinoma. It could be a potential cancer biomarker of prognosis and targeted therapy in OSCC.

Prognostic value of plasma miR-1290 expression in patients with oral squamous cell carcinoma

To investigate the prognostic value of plasma miR-1290 in patients with oral squamous cell carcinoma (OSCC). Seventy patients with OSCC admitted to Danzhou People's Hospital from January 2014 to December 2018 were included in this study. Real-time fluorescence quantitative polymerase chain reaction (PCR) was used to detect the expression of miR-1290 in these patients. The optimal cut-off value of plasma miR-1290 expression was determined by the ROC curve method, and patients with OSCC were divided into the high (n=31) and low (n=39) miR-1290-expressing groups. The clinicopathological features of the two groups were compared, and survival curves were drawn using the Kaplan-Meier method. Risk factors affecting the poor prognosis of patients were analyzed using univariate and multivariate COX regression models. The expression level of plasma miR-1290 in the OSCC group was significantly lower than that in the control group (0.65±0.14 vs. 2.06±0.90; t=13.912, P<0.001). The low expression of plasma miR-1290 appeared to be related to the clinical stage, differentiation degree, tumor diameter, and lymph node metastasis of OSCC (P<0.05). Survival analysis showed that the overall survival rate and the progression-free survival rate of the low-miR-1290 group were significantly lower than that of the high-miR-1290 group (P<0.01). Multivariate COX regression analysis showed that clinical stage, lymph node metastasis, and plasma miR-1290<1.14 were independent risk factors for the poor prognosis of patients with OSCC (P<0.05). The expression level of plasma miR-1290 in patients with OSCC significantly decreased, and the low expression of miR-1290 is related to the short survival time of OSCC patients. Thus, miR-1290 may be a potential marker predicting the poor prognosis of OSCC.

COMPARISON OF EXPRESSION OF E-CADHERIN IN ORAL PSEUDOEPITHELIOMATOUS HYPERPLASIA AND ORAL SQUAMOUS CELL CARCINOMA

Background: Pseudo-epitheliomatous Hyperplasia (PEH) is a benign proliferation of epithelium occurring in response to various neoplastic lesions, infections and inflammatory processes. It can be mistaken especially in small biopsies for an invasive oral squamous cell carcinoma (OSCC). The objective of this study was to determine the expression and comparison of immunohistochemical marker E-cadherin in OSCC and PEH lesions. Materials &amp; Methods: This cross-sectional study was conducted in Department of Histopathology, Armed Forces Institute of Pathology, Rawalpindi, Pakistan from January 2013 to March 2016. 60 paraffin embedded cases, 30 each of PEH and OSCC were retrieved and stained with haematoxylin and eosin. They were then immune-stained with E-cadherin and expression evaluated and compared in two lesions by histopathologist. Age, sex, site of lesion and E-cadherin expression were variables. Age was described by mean and SD and other variables as frequency and percentages. Results: Mean age of OSCC group was 60.1±17.3 years and PEH group 52.7±16.6 years. OSCC group included 16 (53.33%) men and 14 (46.67%) women. PEH group included 18 (60%) men and 12 (40%) women. In OSCC group, site of lesion was buccal mucosa 12 (40%) cases, gingiva 10 (33.3%), tongue 7 (23.3%) and floor of mouth 1 (3.4%) case. In PEH group, site of lesion was buccal mucosa 12 (40%) cases, tongue 11 (36.67%), gingiva 6 (20%) and palate 1 (3.3%) case. The expression of E-cadherin was negative in all 30 cases of OSCC and positive in 29/30 (96.67%) cases in PEH. Conclusion: E-cadherin can be used as an ancillary marker in the differentiation of oral squamous cell carcinoma and pseudo-epitheliomatous hyperplasia.

CYP2 C9 polymorphism among patients with oral squamous cell carcinoma and its role in altering the metabolism of benzo[a]pyrene

The aim of this study was to evaluate the prevalence of CYP2 C9 polymorphism among healthy controls and patients with oral squamous cell carcinoma (OSCC) and to analyze the risk of disease development. We also investigated the interaction between CYP2 C9 wild type and the polymorphic variants with benzo[a]pyrene by using molecular docking analysis. The study included 46 patients with OSCC and 46 controls. Amplification of the genomic DNA was done by using allele-specific polymerase chain reaction and then analyzed by using agarose gel electrophoresis. Molecular docking was then carried out to determine the interaction of CYP2 C9*1, CYP2 C9*2, and CYP2 C9*3 with benzo[a]pyrene. In the OSCC group, CYP2 C9*2 and CYP2 C9*3 polymorphisms were 17.4% and 15.2%, respectively, and in the control group, they were 8.7% and 6.5%, respectively. The OSCC group showed a statistically significant (P=.043) increase in the prevalence of CYP2 C9 polymorphic variants compared with the control group. The docking analysis showed benzo[a]pyrene to bind specifically to the altered single nucleotide catalytic site in the polymorphic CYP2 C9*3 enzyme. This study demonstrates that functionally important CYP2 C9 polymorphism exists among patients with OSCC, with a modest increase in the risk of disease development in those individuals who acquire these poor metabolizing variants. The modified docking of CYP2 C9*3 with benzo[a]pyrene signifies altered metabolism in vivo.

The Efficacy of Squamous Cell Carcinoma Antigen in the Differential Diagnosis of Oral squamous Cell Carcinoma.

Abstract Background and Objectives: Oral squamous cell carcinoma (OSCC) is a major oral cancer representative. Squamous cell carcinoma antigen (Scc-AG) is detected in squamous cell carcinoma of oral tissues and other different tissues. Oral lichen planus (OLP) is a chronic inflammatory disease with questionable malignant transformation. Methods: Forty-five participants divided into OSCC group and two control groups: oral lichen planus as an inflammatory disease (positive control) and healthy volunteers (negative control). Clinical staging and grading of OSCC was recorded. A double-antibody sandwich enzyme-linked immunosorbent assay was used to evaluate the level of Scc-AG in serum and saliva. Results: Statistically significant difference for serum and salivary Scc-Ag, was found between the groups; OSCC, OLP on one hand and healthy controls on the other hand. No statistically significant difference was found between OSCC and OLP control groups in serum levels, while for salivary Scc-Ag a statistically significant difference was found between these two groups, with the higher values in OLP. Staging and grading in OSCC group were not found to be related to Scc-Ag level. Conclusion: Serum Scc-Ag cannot be used as a biomarker to differentiate between oral malignant and chronic inflammatory lesions. On the other hand, salivary Scc-Ag determination provides results more reliable than serum Scc-Ag and could be used in monitoring inflammatory oral disease activity as well as malignant conditions and could differentiate between them.

Ki67, CD105, and α-SMA Expression Supports Biological Distinctness of Oral Squamous Cell Carcinoma Arising in the Background of Oral Submucous Fibrosis.

Background:The clinicopathological distinctness of oral squamous cell carcinoma arising in the background of oral submucous fibrosis (OSCC-OSF) is well known; however, the molecular distinctness of this unique OSCC-OSF has not been investigated to date. With this in mind, we compared the expression of Ki67, CD105, and α-SMA between OSCC-OSF and oral squamous cell carcinoma (OSCC). Methods:Formalin-fixed paraffin-embedded tissues of 105 OSCC-OSF and 112 OSCC cases were subjected to immunohistochemistry for evaluation of Ki67, CD105, and α-SMA expression. Results: Ki67 (labeling index) LI, MVD and α-SMA expression were significantly higher in OSCC compared to OSCC-OSF. Ki67 LI and MVD was significantly higher in OSCC compared to OSCC-OSF in parameters such as well-differentiated, early TNM stage, non-metastatic, and more than 3-year survival. α-SMA expression was significantly higher in OSCC compared to OSCC-OSF in parameters such as moderate differentiation, metastatic lesions, and survival less than 3 years. Ki67 LI, MVD and α-SMA showed significant positive correlation with each other in OSCC and OSCC-OSF. Conclusion:Proliferation, neoangiogenesis and myofibroblast differentiation were significantly higher in the OSCC group compared to the OSCC-OSF group. This suggests the biological distinctness of OSCC-OSF, which could help the future development of targeted therapies.

Expression of DNA repair genes in oral squamous cell carcinoma using reverse transcription-quantitative polymerase chain reaction

The aim of this study was to evaluate the expression of DNA repair genes in cases of oral squamous cell carcinoma (OSCC). Expression of the MLH1, MSH2, MLH3, ATM, MRE11A, XRCC1, and PMS2 genes was evaluated by reverse transcription-quantitative polymerase chain reaction in the OSCC group (32 patients) and the control group (15 patients). The groups were compared by using the Mann-Whitney test, with Bonferroni correction. Associations between gene expression levels and clinical data were explored by using Pearson's and Spearman's correlation coefficients, with P value less than .05 indicating a significant difference. The MLH1, MSH2, MLH3, ATM, MRE11A, XRCC1, and PMS2 genes were downregulated in the OSCC group compared with the control group, with significant values for MLH1 (P < .0001); MSH2 (P=.038); MLH3 (P < .0001); ATM (P < .0001); MRE11A (P < .0001); XRCC1 (P=.0004); and PMS2 (P=.008). Analysis of the correlation between gene expression and clinical data only revealed a significant negative correlation between age and expression of the PMS2 gene. Expression of the DNA repair genes MLH1, MSH2, MLH3, ATM, MRE11 AMRE11A, XRCC1, and PMS2 was reduced in OSCC.

Hypermethylated PAX1 and ZNF582 genes in the tissue sample are associated with aggressive progression of oral squamous cell carcinoma.

DNA methylation of paired box gene 1 (PAX1) and zinc finger 582 (ZNF582) is promising cancer biomarkers for oral squamous cell carcinoma detection. This study aims to investigate the correlation between PAX1 or ZNF582 methylation and the progression of oral squamous cell carcinoma (OSCC). A total of 135 OSCC cases from Peking University School and Hospital of Stomatology were enrolled in this study. Tissue specimens were collected from the lesion site and corresponding adjacent normal site. The methylation level of these two genes was evaluated in primary and recurrent OSCC group. Hypermethylation of PAX1 or ZNF582 was observed in lesion sites among primary and recurrent OSCC cases. In the lesion site of primary cases, promoter methylation was observed in T3/T4 (PAX1: P=.02; ZNF582: P=.01), stage III/IV (PAX1: P=.03; ZNF582: P=.01), and bone invasion cases (PAX1: P=.02; ZNF582: P=.047). In the subgroup analysis, the correlation between hypermethylation and OSCC severity remains significant with exposure to smoking/alcohol consumption. Hypermethylated PAX1 and ZNF582 can sufficiently act as biomarkers to reflect the severity or progression of OSCC.

Prevalence of herpes simplex virus I, II, cytomegalovirus, Epstein–Barr virus and human papillomavirus (6, 11, 16 and 18) in normal oral mucosa and OSCC patients in Saint-Petersburg (Russia).

e18539 Background: The objective of our study was to evaluate the prevalence of DNA of HSV I, II, CMV, EBV, HPV6.11, HPV16, HPV18 in normal oral mucosa and oral squamous cell carcinoma (OSCC), and determine this association with long-term treatment outcomes and patient survival. Methods: The study included 116 patients with OSCC (stage I-IVB who received standard treatment) and 50 healthy volunteers, median age 60.0 yrs (95% CI 58.0-64.0). Using real-time PCR (polymerase chain reaction), freshlyfrozen samples of the tumor and normal mucous membranes of the cavity were analyzed. mouths, which, after collection, were placed in an EDTA solution and frozen at 20°C. Results: In 54 (46.55%) OSCC and 17 (34.00%) normal mucous viral DNA was not found (p = 0.13). The occurrence of DNA (OSCC vs. normal) of the following viruses was the same in both OSCC and normal mucosa: HSV I, II: 2 (1.72%) vs. 2 (4.00%) (p = 0.38); EBV: 40 (34.48%) vs. 15 (30.00%) (p = 0.57); CMV: 5 (4.31%) vs. 2 (4.00%) (p = 0.93); HPV6.11: 16 (13.79%) vs. 6 (15.00%) (p = 0.84); HPV16: 12 (10.34%) vs. 2 (4.00%) (p = 0.18). HPV18 DNA: 20 (17.24%) vs. 24 (48.0%) (p &lt;0.0001) was found significantly less frequently in the OSCC group. In addition, EBV + HPV18: 7 (14.00%) vs. 5 (4.31%) (p = 0.0270) was more common vs. OSCC in the group of healthy volunteers; CMV + HPV18: 2 (4.00%) vs. 0 (p = 0.0302). In the OSCC group, patients with stage III disease were more often virus-positive (p = 0.0137); the pterygo-maxillary fold region (p = 0.0438); with partial keratinization of cells (p = 0.05). PFS in the HPV18 (+) OSCC group was 38.5 months (95% CI 11.00-66.0) and 23.5 m higher than the group of HPV18 (-) patients (HR = 0.63; 95% CI 0.39-1.03; p = 0.0399). In virus-negative patients, PD was more often recorded: 12 (24.00%) vs. 2 (3.03%) (p = 0.0006). In the group of virus-positive patients, OS and PFS, even with non-radical treatment, were significantly higher than virus-negative patients with radical treatment. OS: vir (+)/non-radical = 69.0 mo vs. vir (-)/radical = 15.0 mo (HR = 0.22; 95% CI 0.11-0.47; p &lt;0.0001). PFS: vir (+)/non-radical = 56.0 mo vs. vir (-)/radical = 11.0 mo (HR = 0.20; 95% CI 0.09-0.45; p &lt;0.0001). Conclusions: The association of OSCC with viral DNA, especially HPV18, is a favorable prognosis of the disease and a longer OS and PFS, even when radical treatment is not possible.

Alternation of β-catenin and CD44s Immunoexpression in Different Histopathological Grades of Oral Squamous Cell Carcinoma.

Objective:Cell-cell adhesion molecules play an essential role in cell growth and differentiation. β-catenin and CD44s are two adhesion molecules which their expression changes are correlated to loss of differentiation and gain of an invasive epithelial phenotype. Oral squamous cell carcinoma (OSCC) is the most common malignancy of oral cavity. Aim of this study was to compare β-catenin and CD44s expression in different histopathological grades of OSCC. Methods:β-catenin and CD44s expression were evaluated in 10 well differentiated OSCC (group A) and 10 moderately/poorly differentiated OSCC (group B) using immunohistochemistry. Results:β-catenin membranous and nuclear/cytoplasmic expression were significantly different between groups A and B. CD44s membranous expression was insignificant amongst the two groups. Conclusion:Expression of β-catenin and CD44s alter in different histopathological grades of OSCC. It seems that more rate of aberrant cytoplasmic and/nuclear expression and less membranous expression of β-catenin can lead to significantly lower degree of cell differentiation in OSCC.

Study on expression of LIM domain only protein 1 in SD rat oral buccal mucosa carcinogenesis induced by 4-nitro-quinoline N-oxide

This work aimed to determine the expression changes in LIM domain only protein 1 (LMO1) in gene transcription and protein levels during oral squamous cell carcinoma (OSCC) development. The tissues in this study were taken from our team's previous animal model building, and we performed hematoxylin-eosin (HE) staining on 49 cases. The pathological classification of the experiment group was determined on the basis of the abnormal epithelial hyperplasia degree. The expression part of LMO1 was determined by immunohistochemistry staining. The mRNA and protein LMO1 expression levels in five groups were detected by real-time fluorescent quantitative of nucleotide polymer chain reaction (RT-qPCR) and Western blot, respectively. HE staining determined 7 cases of the control group, 6 cases of mild epithelial dysplasia, 11 cases of moderate epithelial dysplasia, 9 cases of severe epithelial dysplasia, and 16 cases of OSCC. Immunohistochemistry results: LMO1 expression was localized in the cytoplasm, and the positive expression rates of the protein LMO1 in the control and experiment groups were 14.3% for normal buccal mucosal tissue, 33.3% for mild epithelial dysplasia, 81.8% for moderate epithelial dysplasia, 88.9% for severe epithelial dysplasia, and 93.8% for OSCC. RT-qPCR results: mRNA expression was lowest in the control group and highest in the OSCC group, the difference between the mild dysplasia and control groups was not significant (P>0.05). Pairwise comparison among other groups showed statistically significant differences (P<0.05). Western blot results: with the aggravation of the pathological degree, the protein LMO1 expression level increased gradually. The OSCC group expressed the highest LMO1 expression level. The oral mucosa carcinogenesis models showed abnormal the mRNA and protein LMO1 expression levels, and the mRNA and protein expression levels were positively correlated with the degree of abnormal proliferation.

EXPRESSION OF MATRIX METALLOPROTEINASE-9 IN ORAL SQUAMOUS CELL CARCINOMA AND ORAL PSEUDOEPITHELIOMATOUS HYPERPLASIA

Background: Oral pseudoepitheliomatous hyperplasia (PEH) appears histologically similar to oral squamous cell carcinoma (OSCC) in small oral biopsies, thus posing diagnostic dilemma. The objective of this study was to compare the expression of matrix metalloproteinase-9 (MMP-9) in differential diagnosis of OSSC and oral PEH. Materials &amp; Methods: This comparative cross-sectional study was conducted in the Department of Histopathology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, Pakistan from January 2013 to March 2016. It included sixty archival cases, 30 each of OSCC and PEH. Paraffin embedded blocks were prepared, hematoxylin and eosin stained sections taken and immunostained with MMP-9. The expression of MMP-9 was evaluated in OSCC and PEH. Results: The OSCC group included 16 (53.33%) men and 14 (46.67%) women, whereas PEH group included 18 (60%) men and 12 (40%) women. The mean age of OSCC group was 60.1±17.3 and that of PEH group was 52.7±16.6. In OSCC group, site of lesion was buccal mucosa in 12 (40%), gingiva 10 (33.33%), tongue 7 (23.33%) and floor of mouth 1 (3.34%) case. In PEH group, site of lesion was buccal mucosa in 12 (40%), tongue 11 (36.66%), gingiva 6 (20%) and palate 1 (3.34%) case. The expression of MMP-9 was positive in all the 30 cases of OSCC and negative in all 30 cases of PEH. Conclusion: Compared to pseudoepitheliomatous hyperplasia (PEH), MMP-9 revealed a higher expression in oral squamous cell carcinoma (OSCC). This finding has become mainstream strategy in distinguishing OSCC from PEH in oral mucosal biopsies in cases difficult to diagnose.

IMMUNOHISTOCHEMICAL EVALUATION OF LAMININ-5 AND MYOFIBROBLASTS IN EPITHELIAL DYSPLASIA AND ORAL SQUAMOUS CELL CARCINOMA

Background: Oral squamous cell carcinoma is one of the most prevalent cancers worldwide. The transformation of the normal epithelial cell into a tumor cell bestows upon them certain features at the cellular and molecular level which aids in its survival and proliferation. Invasion of the altered tumor cells through the basement membrane into the connective tissue stroma and their subsequent spread and metastasis is an important prognostic indicator. Laminin-5 is a protein associated with a migratory phenotype in epithelial neoplastic cells. Along with laminin, the stromal myofibroblasts play a significant role in tumor invasion, due to its ability to modify the extracellular matrix.&#x0D; Aim: To evaluate the role of laminin 5 and stromal myofibroblasts in oral epithelial dysplasia and oral squamous cell carcinoma.&#x0D; Methods: Paraffin-embedded archival samples of 25 normal, 30 oral epithelial dysplasia and 30 oral squamous cell carcinoma (OSCC) were evaluated for laminin-5 and α- smooth muscle actin (SMA) using standard immunohistochemistry. Semi-quantitative assessment of the expression of laminin and alpha SMA was done in all the study samples. The area of staining and the staining intensity was evaluated in order to determine the staining index which were then statistically analyzed between the three groups.&#x0D; Results: All the cases of laminin showed cytoplasmic staining in the basal cell layer and basement membrane. Expression of laminin was observed in the basal cell layer of normal and epithelial dysplasia study group and mainly around the tumor islands in OSCC group. α- SMA was seen with increasing intensity with increasing grade of the disease. Comparison of laminin expression between the three groups showed a statistically significant decrease in the staining index from normal to epithelial dysplasia to OSCC (p &lt; 0.01). Statistical comparison of α-SMA in between the three groups using Kruskal- Wallis test showed a significant increase in the expression of α-SMA from normal to epithelial dysplasia to OSCC (p &lt; 0.01)&#x0D; Conclusion: Decreased laminin expression in the basement membrane and increased expression of α-SMA favors tumor invasion, establishment of an invasive phenotype of neoplastic cells and a permissive environment for tumor invasion.&#x0D; Key words: Epithelial dysplasia, Oral cancer, Laminin, Alpha- Smooth muscle actin, Immunohistochemistry, Myofibroblasts

Potential Salivary mRNA Biomarkers for Early Detection of Oral Cancer.

We evaluated potential biomarkers in human whole saliva for the early diagnosis of oral squamous cell carcinoma (OSCC). We selected 30 candidate genes with relevance to cancer from recent reports in PubMed. Saliva samples were obtained from 34 non-tumor control and 33 OSCC patients. Real-time PCR was performed, and mRNA levels were compared. Normalized mRNA levels of six genes (NGFI-A binding protein 2 (NAB2), cytochrome P450, family 27, subfamily A, polypeptide 1 (CYP27A1), nuclear pore complex interacting protein family, member B4 (NPIPB4), monoamine oxidase B (MAOB), sialic acid acetyltransferase (SIAE), and collagen, type III, alpha 1 (COL3A1)) were significantly lower in saliva of OSCC patients. Receiver operating characteristics (ROC) analysis was used to individually evaluate the predictive power of the potential biomarkers for OSCC diagnosis. The area under the curve (AUC) values were evaluated for the OSCC vs. non-tumor groups via univariate ROC analyses, as well as multivariate ROC analyses of combinations of multiple potential biomarkers. The combination of CYP27A1 + SIAE showed a favorable AUC value of 0.84. When we divided saliva samples into two groups according to age using a 60-year cut-off, with OSCC patients and controls evaluated together, the AUC of MAOB–NAB2 was more predictive of OSCC in the under-60 group (AUC, 0.91; sensitivity, 0.92; and specificity, 0.86) than any other gene combination. These results are expected to aid the early diagnosis of OSCC, especially in patients under 60 years of age. While more studies with larger numbers of patients are necessary, our result suggest that salivary mRNA would be a potent biomarker for early OSCC diagnosis.

The Oral Microbiota May Have Influence on Oral Cancer.

The oral microbiota plays an important role in the human microbiome and human health, and imbalances between microbes and their hosts can lead to oral and systemic diseases and chronic inflammation, which is usually caused by bacteria and contributes to cancer. There may be a relationship between oral bacteria and oral squamous cell carcinoma (OSCC); however, this relationship has not been thoroughly characterized. Therefore, in this study, we compared the microbiota compositions between tumor sites and opposite normal tissues in buccal mucosal of 50 patients with OSCC using the 16S rDNA sequencing. Richness and diversity of bacteria were significantly higher in tumor sites than in the control tissues. Cancer tissues were enriched in six families (Prevotellaceae, Fusobacteriaceae, Flavobacteriaceae, Lachnospiraceae, Peptostreptococcaceae, and Campylobacteraceae) and 13 genera, including Fusobacterium, Alloprevotella and Porphyromonas. At the species level, the abundances of Fusobacterium nucleatum, Prevotella intermedia, Aggregatibacter segnis, Capnocytophaga leadbetteri, Peptostreptococcus stomatis, and another five species were significantly increased, suggesting a potential association between these bacteria and OSCC. Furthermore, the functional prediction revealed that genes involved in bacterial chemotaxis, flagellar assembly and lipopolysaccharide (LPS) biosynthesis which are associated with various pathological processes, were significantly increased in the OSCC group. Overall, oral bacterial profiles showed significant difference between cancer sites and normal tissue of OSCC patients, which might be onsidered diagnostic markers and treatment targets. Our study has been registered in the Chinese clinical trial registry (ChiCTR1900025253, http://www.chictr.org.cn/index.aspx).