Specimens Of Normal Oral Mucosa Research Articles

Immuno-histochemical evaluation of CD34 for OLP and OSMF

Immuno-histochemical evaluation of CD34 in oral lichen planus (OLP) and Oral Submucous Fibrosis (OSMF) is of interest to dentist.20 specimens of normal oral mucosa (buccal mucosa/gingiva tissue) from patients who had extractions performed as part of orthodontic treatment comprised Group I, the control group. Group II comprised 30 individuals with a diagnosis of oral lichen planus. 30 OSMF cases with diagnoses is Group III. These 80 specimens were all given consideration when choosing for CD34immuno-histochemical staining. The CD34 was greater in all categories of OLP and OSMF when compared to normal controls. Maximum CD34 expression was observed in erosive OLP (147.41ą17.60) followed by OSMF (116.01 ą11.72) and reticular OLP (105.01ą11.62). Data was statistically significant (p<0.001).Immunohistochemistry of CD34 evaluation is a potential diagnostic marker for OLP and OSMF.

Expression analysis of cyclin D, Ki-67, MCM3 and MCM2 in oral squamous cell carcinoma.

The expression analysis of cyclin D1, Ki-67, MCM3 and MCM2 in oral squamous cell carcinoma to identify biomarkers is of interest. 45 formalin-fixed paraffin embedded tissue blocks collected from archives of Department of Oral and Maxillofacial Pathology and Oral Microbiology, Government Dental College and Hospital, Jamnagar, India were subjected to a retrospective cross-sectional immuno-histo-chemistry examination. 30 blocks of OSCC with histological diagnosis have 15 tissue blocks of well-differentiated oral carcinoma and 15 tissue blocks of moderately-differentiated oral carcinoma. 15 specimens of normal oral mucosa (NM) were also examined for comparison. In each of the categories, the immuno-histo-chemistry expression of cyclin D1, MCM 3, MCM 2, and ki67 was studied. Data shows that cyclin D1, Ki-67, MCM3 and MCM2 effectively indicate cellular proliferation for consideration as potential biomarkers of oral squamous cell carcinoma.

Immunoexpression of HSP27 does not seem to influence the prognosis of oral tongue squamous cell carcinoma.

New methods of early detection and risk assessment have been studied aiming to predict the prognosis of patients and directing a specialized treatment of the oral tongue squamous cell carcinoma (OTSCC). In this context, several molecular biomarkers have been investigated for this purpose, and, among them, the heat shock protein 27 (HSP27) can be named. The study aimed to analyze whether heat shock protein 27 (HSP27) exerts any influence on OTSCC, correlating its immunoexpression with clinicopathological parameters, and patient survival. The sample comprised 55 OTSCC cases and 20 normal oral mucosa specimens. The malignancy grading systems proposed by the WHO in 2005, Brandwein-Gensler et al., and Almangush et al. were applied in a histomorphological study. HSP27 expressions were evaluated through the Immunoreactivity Score System (IRS). Significant values were considered at p <0.05 for all statistical tests. Higher IRS results were observed for normal oral mucosa specimens when compared to OTSCC cases (p <0.001). No significant associations between HSP27 immunostaining, the analyzed clinicopathological parameters and patient survival were observed. The results of the present study indicate lower HSP27 expression in OTSCC cases compared to normal oral mucosa specimens. Thus, HSP27 expression does not seem to influence patient prognosis.

Lymphatic Vascular Density, the Expression of Podoplanin and Tumor Budding in Oral Squamous Cell Carcinoma.

Notwithstanding recent advances in oral squamous cell carcinoma (OSCC) management, its mortality rate is still high. It is imperative to investigate new parameters that are complementary to clinical staging for OSCC to provide better prognostic insight. The presence of isolated neoplastic cells or small clusters of up to four cells at the tumor's invasive front, called tumor budding, is a morphological marker of OSCC with prognostic value. Increased lymphatic vascular density (LVD) and a high expression of podoplanin in neoplastic cells have also been associated with worse prognosis in OSCC. To investigate these markers in OSCC, we evaluated differences in LVD and the expression of podoplanin in neoplastic cells between tumors with high-intensity tumor budding versus low-intensity or no tumor budding. In the samples of high-intensity budding, differences in those parameters between the​​ budding area and the area outside the budding were also evaluated. Furthermore, the study assessed differences in LVD and in the expression of podoplanin in neoplastic cells concerning OSCC clinicopathological characteristics. To those ends, we subjected 150 samples of OSCC to immunohistochemistry to evaluate the intensity of tumor budding (via multi-cytokeratin immunostaining). Moreover, the 150 samples of OSCC and 15 specimens of normal oral mucosa (used as a control) were employed to assess LVD and the expression of podoplanin (in neoplastic cells of OSCC and in the lining epithelium of normal oral mucosa), both via podoplanin immunostaining. Data were processed into descriptive and analytical statistics. No differences were observed neither in the LVD nor in the expression of podoplanin in neoplastic cells concerning sex, age, tobacco smoking, tumor location and tumor size. The LVD was greater in OSCC and in tumors with high-intensity budding than in normal mucosa but did not differ between normal mucosa and tumors with low-intensity or no tumor budding. The data analyses also revealed that LVD was greater in tumors with high-intensity tumor budding than in tumors with low-intensity or no budding and showed no difference in LVD between the budding area and the area outside the budding. When compared to the lining epithelium of the normal mucosa, the expression of podoplanin was greater in neoplastic cells of OSCC, tumors with high-intensity budding and tumors with low-intensity or no tumor budding. The expression of podoplanin in neoplastic cells was also greater in tumors with high-intensity budding and, within those tumors, greater in the budding area than in the area outside de budding. Those findings support the hypothesis that tumor budding is a biological phenomenon associated with the progression and biological behavior of OSCC.

Combined evaluation of both WEE1 and phosphorylated cyclin dependent kinase 1 expressions in oral squamous cell carcinomas predicts cancer recurrence and progression

Background/purposeWEE1 is a mitotic inhibitor at G2 checkpoint of the cell cycle that negatively regulates cyclin-dependent kinase 1 (CDK1) through inhibitory phosphorylation. This study assessed whether the expressions of both WEE1 and phosphorylated CDK1 in specimens of oral squamous cell carcinoma (OSCC) might predict the OSCC recurrence and progression. Materials and methodsThis study used immunohistochemistry to examine the expressions of WEE1 and phosphorylated CDK1 proteins in 75 specimens of OSCC and 30 specimens of normal oral mucosa (NOM). ResultsThe mean WEE1 labeling index (LI) was significantly lower in 75 OSCC samples than in 30 NOM samples (P < 0.001), whereas the mean phosphorylated CDK1 LI was significantly higher in 75 OSCC samples than in 30 NOM samples (P < 0.001). We found a significant association of low WEE1 LI (<21%) with OSCC recurrence (P = 0.047) and a significant association of low phosphorylated CDK1 LI (<10%) with larger tumor size (P = 0.011) and more advanced clinical stages (P = 0.021) of OSCC. ConclusionCombined evaluation of WEE1 and phosphorylated CDK1 LI in specimens of OSCC may predict the OSCC recurrence and progression.

Immunohistochemical Expression of Cyclin D1 in Oral Lichen Planus

Background: Many studies have reported that a condition initially classified as oral lichen planus (OLP) has a variable chance of developing cancer progression over time, although these results are still questionable. Cyclin D1 controls the mitotic cell cycle's transition from G1 to S. Cyclin D1 has a significant function in carcinogenesis due to its critical involvement in cell cycle control. Cyclin D1 deregulation or overexpression may result in shortened G1 phase, enhanced cell replication, and decreased reliance on growth regulators, causing disruption in conventional cell cycle progression and the progression of cancer. The aim of the current study was to evaluate the expression of Cyclin D1 in histopathologically confirmed OLP samples and to predict the rate of malignant transformation in these lesions by comparing the level of expression with some clinical parameters. Materials and Methods: The immunohistochemical procedure was used to examine the expression of cyclin D1 on paraffin-embedded sections of (40) OLP lesions and (10) specimens of normal oral mucosa. We used the SPSS software version (18.0) to do the statistical analysis in this work. Results: There is a very significant difference between the people investigated (P = 0.01), which is statistically significant. CyclinD1 was found in all of the oral lichen planus cases in the research (100%), but it was also found (80%) in the normal oral mucosa. There was also no statistically significant difference between the clinical types of OLP and the Cyclin D1 scoring (P = 0.942), which revealed that the reticular type was the most frequent (7) in mild scoring, whereas the erosive type was the most frequent in severe scoring, which accounted for the majority of the cases. Conclusion: This research found that higher cyclinD1 expression increases the risk of cancer and many of the markers should be tested. Furthermore, larger sample numbers, improved designs, and gene-reduction studies should be performed. OLP patients should be continuously monitored for OSCC progression.

Cyclooxygenase-2 (COX-2) Expression in Oral Submucous Fibrosis and Oral Squamous Cell Carcinoma: An Immunohistochemical Study.

Introduction:Growing evidence has shown that cyclooxygenase-2 (COX-2), an enzyme capable of catalyzing prostaglandin production, plays a key role in carcinogenesis. Selective COX-2 inhibitors have been shown to reduce the establishment of tumors such as oral squamous cell carcinoma (OSCC) and premalignant conditions such oral submucous fibrosis (OSMF) in experimental models. The aim of this study was to investigate the immunohistochemical expression of COX-2 in OSCC and OSMF with the normal oral mucosa as control.Material and Methods:Forty-five formalin-fixed paraffin-embedded samples comprising 20 OSCC, 20 OSMF, and 5 normal oral mucosa specimens were withdrawn from the archives of the Department of Oral and Maxillofacial Pathology for immunohistochemical examination for COX-2 expression. Negative and less than 5% COX-2 positivity was considered negative expressions, while greater than or equal to 5% COX-2 positivity was considered positive expression. The data obtained were statistically analyzed.Results:The difference in percentages of expression in normal mucosa, OSCC, and OSMF was highly significant (P < 0.01). In comparison to normal mucosa, OSCC and OSMF had an increased level of COX-2 expression. However, there was an insignificant difference between the various histological gradings of OSCC and OSMF.Conclusion:The results of the present study confirm the role of COX-2 in carcinogenesis and in the progression of premalignant conditions to malignancy.

Melanoma Inhibitory Activity and Melanoma Inhibitory Activity 2 as Novel Immunohistochemical Markers of Oral Epithelial Dysplasia.

Oral potentially malignant disorders are associated with the development of oral squamous cell carcinoma (OSCC). Most OSCCs are diagnosed via histopathology as oral epithelial dysplasia (OED), but the histologic diagnostic criteria remain non-uniform. Accordingly, the establishment of a diagnostic marker to assist in diagnosis could contribute towards cancer prevention. Melanoma inhibitory activity (MIA) and MIA2 are involved in tumor growth, invasion, and lymph node metastasis in various malignancies. The purpose of this study was to clarify the usefulness of MIA and MIA2 as diagnostic markers of oral mucosal lesions. The expression of MIA and MIA2 was analyzed immunohistochemically in 100 specimens (10 specimens with normal oral mucosa (NOM) and 30 specimens each with low-grade epithelial dysplasia (LED), high-grade epithelial dysplasia (HED), and OSCC). Immunohistochemical results were evaluated based on the Allred scoring system. Cytoplasmic expression of MIA and MIA2 increased in the order of LED, HED, and OSCC. All NOM specimens were negative for cytoplasmic expression. Significant differences were observed between the groups (NOM vs. HED, p < 0.05, NOM vs. OSCC, p < 0.001). These results demonstrate that MIA and MIA2 are expressed in the oral mucosa within early neoplastic lesions and suggest that MIA and MIA2 are useful novel immunohistochemical markers for discriminating between normal tissue and OED.

Expression of transmembrane protein aquaporin-3 in oral epithelial dysplasia and oral squamous cell carcinoma

The objective of this study was to evaluate aquaporin-3 (AQP3) expression in patient samples of oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC), thereby assessing the potential of AQP3 as a molecular marker for tumor progression. An in vitro comparative study was done to determine the AQP3 expression on 20 surgical biopsy specimens each of OED and OSCC using immunohistochemistry. Twenty specimens of normal oral mucosa were kept as controls. The results were statistically analyzed using one-way analysis of variance and post hoc analysis. The expression of AQP3 was analyzed and further semiquantified using H-scores. The mean H-score showed a statistically significant difference between OSCC, OED, and normal oral mucosa (P < .05). There was a significant increase in the expression of AQP3 in OSCC and OED compared to normal oral mucosa. The highest expression was observed in OSCC (P < .01). The observations of the study indicate that staining intensity of AQP3 increased from dysplastic noninvasive lesion to invasive OSCC, suggesting a possible role of AQP3 as a biomarker for tumor progression.

Detection of Micro-invasion in Sudanese Oral Verrucous Carcinoma Samples Using Syndecan-1 Stain.

Aim:Verrucous carcinoma (VC) is a low-grade rare variant of squamous cell carcinoma (SCC). Syndecan-1 (CD138) is a heparan sulfate proteoglycan which participates in cell-to-cell adhesion and cell-matrix interaction. Being misled by the apparent non-aggressive nature of VC, some clinicians and pathologists believe that this tumor is not an aggressive tumor, not realizing the fact that some of these lesions may contain nests or foci of well-differentiated SCC. This study aimed to assess syndecan-1 expression of VC and detection of micro-invasion in VC using syndecan-1 immunohistochemical (IHC) stain.Methods:Observational analytical study of 34 paraffin block of VC cases and 24 cases of variable grades of oral epithelial dysplasia. Cases were stained by hematoxylin and eosin (H&E) and then IHC stain for syndecan-1 was applied. Nine paraffin blocks from specimens of normal oral mucosa were used as the reference group for syndecan-1 stain positivity.Results:In this study, we found that 32 (94.1%) out of 34 of verrucous carcinoma cases showed loss of syndecan-1 expression. Moreover, highly statistically significant association was found between the presence of suggestive micro-invasion in H&E and loss of syndecan-1 expression in micro-invasive area in the same case.Conclusions:In conclusion, syndecan-1 stain can be used as a biomarker in detection of micro-invasion in verrucous carcinoma.

Study of extrinsic apoptotic pathway in oral Lichen Planus using TNFR 1 and FasL immunohistochemical markers and TUNEL technique

ObjectiveThe exact pathogenesis of lichen planus (LP) is still unknown and there are some controversies concerning role of apoptosis in its creation. The purpose of the present study is to investigate extrinsic apoptotic pathway in oral lichen planus(OLP). MethodsIn the present cross-sectional study, the presence of apoptosis was investigated on 25 specimens of OLP and 6 specimens of normal oral mucosa using TUNEL technique and also pro apoptotic immunohistochemical(IHC) markers of FasL and TNFR1 in 4 areas of degeneration, basal and parabasal layers, and lymphocytic band. Then staining intensity distribution (SID) index was determined and the results were analyzed by Wilcoxon and Mann-Whitney tests (P-value <0.05). ResultsThere was no significant difference between SID of TUNEL in the areas of parabasal and lymphocytic band, but the difference was significant among other areas. There was significant difference between SID of TNFR1 in the areas of degeneration and lymphocytic band, basal and lymphocytic band and also parabasal and lymphocytic band, but there was no significant difference between other areas.There was no significant difference between SID of FasL in the areas of degeneration and basal, degeneration and parabasal, and also basal and parabasal, but there was significant difference in other areas. ConclusionsApoptosis probably doesn't play a major role in destruction of basal cells in OLP. Destruction of basal cells in OLP is probably resulting from necrosis of these cells. Because of low level or lack of apoptosis, there is low probability of carcinomatous changes in OLP.

Langerhans cell counts in oral epithelial dysplasia and their correlation to clinicopathological parameters.

Langerhans cells (LCs) are antigen presenting cells. This study assessed the LC counts in oral epithelial dysplasia (OED) and their correlation to clinicopathological parameters. This study examined the LC counts in the epithelia and subepithelial connective tissues of 58 patients with OED (21 mild, 18 moderate, and 19 severe OED lesions) and 10 specimens of normal oral mucosa (NOM) by anti-S-100 protein immunostaining. We found that the mean LC counts in the epithelia or subepithelial connective tissues increased significantly from NOM samples through mild and moderate OED to severe OED samples. In addition, a significant correlation was found between higher mean LC counts in the dysplastic epithelia of OED samples and OED lesions with thicker epithelial layers (p<0.001) or wider inflammatory zones (p<0.001), and between higher mean LC counts in the subepithelial connective tissues of OED samples and OED lesions with wider inflammatory zones (p<0.001). Moreover, the nine OED lesions with malignant transformation had a significantly lower mean LC count than the 49 OED lesions without malignant transformation. The significant and gradual elevation in LC count from NOM through mild and moderate OED to severe OED lesions suggests an upregulation of immunosurveillance ability in OED patients during the early oral carcinogenesis process. A low LC count in OED lesions may suggest the partial loss of immunosurveillance ability against dysplastic cells; this in turn favors the malignant transformation of an OED lesion into oral cancer.

Expression of survivin and p53 in oral lichen planus, lichenoid reaction and lichenoid dysplasia: An immunohistochemical study.

Context:The malignant transformation potential of oral lichen planus (OLP) and related lesions is a subject of great controversy.Aim:The aim of this study was to compare the expression of proteins related to apoptosis and tumour suppressor gene processes in OLP, oral lichenoid reaction (OLR) and oral lichenoid dysplasia (OLD).Materials and MethodsThe immunohistochemical study was carried out to investigate the expressions of survivin and p53 in a total of 30 lesional biopsy specimens - 10 cases each of OLP, OLR and OLD. The expression rates were further compared with 10 control specimens of normal oral mucosa (NORM).Results:Immunoreactivity for p53 was seen in 7 cases (70%) of OLD, 4 cases (40%) of OLP and 2 cases (20%) of OLR and none of NORM. We obtained a significant difference (P = 0.01) in mean p53 expression between the different entities. The positive staining rate of survivin was found to be significantly different between OLD (50%), OLP (10%), OLR (0%), and normal mucosa (0%) (P = 0.004). There was a positive correlation between p53 and survivin expression in OLP and OLD using Pearson's correlation coefficient.Conclusion:Lichenoid dysplasia has shown p53 and survivin expression in the range of not OLP, but leukoplakia. On the other hand, OLR seems to be an innocuous lesion. The study results with OLP are inconclusive but points toward a small but important malignant potential in OLP. This kind of comparative study highlights the importance of biopsying OLP and related lesions for proper diagnosis and appropriate management.

B7-H4 expression indicates poor prognosis of oral squamous cell carcinoma.

Checkpoint blockade therapy utilizing monoclonal antibodies to reactivate T cells and recover their antitumor activity makes an epoch in cancer immunotherapy. The role of B7-H4, a novel negative immune checkpoint, in oral squamous cell carcinoma (OSCC) has still not been elucidated. In this study, tissue samples from human OSCC, which contains 165 primary OSCC, 48 oral epithelial dysplasia and 43 normal oral mucosa specimens, and Tgfbr1/Pten 2cKO mice OSCC model were stained with B7-H4 antibody to analyze the correlations between B7-H4 expression and clinicopathological characteristics. Kaplan-Meier analysis was used to compare the survival of patients with high B7-H4 expression and patients with low B7-H4 expression. We found B7-H4 is highly expressed in human OSCC tissue, and the B7-H4 expression level was associated with the clinicopathological parameters containing pathological grade and lymph node status. Moreover, we confirmed that B7-H4 was overexpressed in Tgfbr1/Pten 2cKO mice OSCC model. Our data also indicated that patients with high B7-H4 expression had poor overall survival compared with those with low B7-H4 expression. Furthermore, this study demonstrated that B7-H4 was positively associated with PD-L1, CD11b, CD33, PI3Kα p110, and p-S6 (S235/236). Taken together, these findings suggest B7-H4 is a potential target in the treatment of OSCC.

Peroxiredoxin 1 promotes invasion and migration by regulating epithelial-to-mesenchymal transition during oral carcinogenesis.

Tobacco smoking is the major risk factor for oral squamous cell carcinoma (OSCC). Previously, we found that nicotine up-regulates peroxiredoxin 1 (Prx1), an important antioxidant enzyme, and nuclear factor kappa B (NFκB) in OSCC cells. However, the molecular mechanism of Prx1 in oral carcinogenesis remains obscure. To improve our understanding of the functional role of Prx1 during the cascade of tobacco-associated oral carcinogenesis, we characterized Prx1, NFκB, and epithelial-to-mesenchymal transition (EMT) markers including E-cadherin, vimentin and Snail in 30 primary oral tumors (15 from smokers with OSCC and 15 from non-smokers with OSCC) and 10 normal oral mucosa specimens from healthy individuals. The expression levels of Prx1, nuclear NFκB, vimentin and Snail were higher in the tumors from smokers with OSCC than in those from non-smokers with OSCC or the healthy controls. The expression levels of E-cadherin showed an opposite trend. Prx1 silencing suppressed the nicotine-induced EMT, cell invasion and migration in SCC15 cells in vitro. Furthermore, Prx1 activated the NFκB pathway in SCC15 cells. Prx1 might therefore play an oncogenic role in tobacco-related OSCC and thus serve as a target for chemopreventive and therapeutic interventions.

CD1a+ dendritic cells in oral lichen planus and amalgam lichenoid reaction

To evaluate and compare the number of CD1a+ dendritic cells (DCs) in oral lichen planus (OLP), amalgam lichenoid reaction (ALR), and normal oral mucosa (NOM). Fourteen cases of OLP and ALR, diagnosed both clinically and histologically, were obtained from an archive of an oral pathology service. Seven specimens of NOM were collected and served as controls. Immunohistochemistry was used to identify CD1a+ DCs. Densities were calculated using specimens of epithelium and lamina propria. For OLP and ALR, specimens of lamina propria included cells from inflammatory infiltrate (region A), and from below inflammatory infiltrate (region B). Comparisons and statistical analyses were performed among OLP, ALR, and NOM. A higher density of CD1a+ DCs was observed in OLP compared to ALR in region A (P<.05) and regions A+B (P<.05). A higher density of CD1a+ DCs was also observed in OLP compared with NOM in the regions A+B (P<.05). Statistically significant differences were not observed for analyses involving epithelial tissue. The observation that there is a lower quantity of DCs in ALR specimens compared with specimens from individuals with OLP suggests that the role for DCs in the 2 diseases may be different. The difference in cell density could be used in the differential diagnosis of these 2 diseases, but the overlapping intervals in the statistical analysis may limit the application of this approach in surgical practice.

Largescale Transcriptomics Analysis Suggests Over-Expression of BGH3, MMP9 and PDIA3 in Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) has been reported as the most prevalent cancer of the head and neck region, while early diagnosis remains challenging. Here we took a comprehensive bioinformatics study on microarray data of 326 OSCC clinical samples with control of 165 normal tissues. The cell interaction pathways of ECM-receptor interaction and focal adhesion were found to be significantly regulated in OSCC samples. Further analysis of the topological properties and expression consistency identified that three hub genes in the gene interaction network, MMP9, PDIA3 and BGH3, were consistently up-expressed in OSCC samples. When being validated on additional microarray datasets of 41 OSCC samples, the validation rate of over-expressed BGH3, MMP9, and PDIA3 reached 90%, 90% and 84% respectively. At last, immuno-histochemical assays were done to test the protein expression of the three genes on newly collected clinical samples of 35 OSCC, 20 samples of pre-OSCC stage, and 12 normal oral mucosa specimens. Their protein expression levels were also found to progressively increase from normal mucosa to pre-OSCC stage and further to OSCC (ANOVA p = 0.000), suggesting their key roles in OSCC pathogenesis. Based on above solid validation, we propose BGH3, MMP9 and PDIA3 might be further explored as potential biomarkers to aid OSCC diagnosis.

Role of human papilloma virus-16 in the pathogenesis of oral lichen planus – an immunohistochemical study

Oral lichen planus (OLP) is a common chronic inflammatory immune-mediated disease with an aetiopathogenesis associated with cell-mediated immunological dysfunction. It is possible that oral mucosal viral infections, including human papilloma virus-16 (HPV-16) infection, may have a causative role in OLP pathogenesis. To assess the prevalence of HPV-16 in histopathologically diagnosed specimens of OLP and to evaluate whether any clinical features (such as the localisation of specimens) or the age or gender of patients, are correlated with the presence of this virus. This study was conducted on 30 specimens with a histopathological diagnosis of OLP, using the immunohistochemical marker HPV-16. Thirty normal oral mucosa specimens were also included as controls. Brown nuclear staining was accepted as positive for the HPV-16 antibody. The results were analysed using Fisher's exact test. P values<0.05 were considered to be significant. Significant correlation (P=0.0001) was observed between HPV-16 infection and samples with OLP. No statistical conclusions could be drawn regarding age, gender, localisation and HPV-16 positivity. Our study showed that HPV-16 may play a role in the pathogenesis of OLP. Taking into account the oncogenic potential of HPV-16, patients with OLP should be screened for the presence of this virus.

Cigarette Smoke Modulates NOD1 Signal Pathway and Human ß Defensins Expression in Human Oral Mucosa

Background/Aims: Nucleotide binding oligomerization domain 1 (NOD1) signal pathway and human ß defensins (hBDs) play crucial roles in innate immune. Cigarette smoke has been confirmed to dampen innate immune in some human tissues, such as oral mucosa. The aim of this study was to evaluate potential effects of smoking on NOD1 signaling and hBDs expression in oral mucosa. Methods: Tissue specimens of normal oral mucosa were collected from donors undergoing routine surgical treatment. All 20 participants were classified equally as two groups: non-smokers and smokers. By using Western blotting and immunohistochemistry, we investigated differential expression of crucial molecules in NOD1 signal pathway, hBD-1, -2, and -3 in oral mucosa tissues between non-smokers and smokers. Immortalized human oral mucosal epithelial (Leuk-1) cells were treated with various concentrations of cigarette smoke extract (CSE) for 24h. Western blotting and immunofluorescence assays were performed to study CSE-induced alteration of protein expression. Leuk-1 cells were treated with 4% CSE, iE-DAP (NOD1 agonist), CSE + iE-DAP, BAY 11-7082 (NF-κB inhibitor), 4% CSE + BAY 11-7082, respectively. Real-time PCR and ELISA were performed to detect the mRNA levels and secretion of hBD-1, -2, and -3, respectively. Results: The levels of NOD1, NF-κB, hBD-1 and hBD-3 significantly reduced in oral mucosa tissues of smokers compared with non-smokers. The levels of RIP2 (receptor-interacting protein 2), phospho-NF-κB (P-NF-κB) and hBD-2 remarkably enhanced in oral mucosal tissues of smokers. CSE treatment suppressed NOD1 and NF-κB expression and activated RIP2 and P-NF-κB expression in Leuk-1 cells. The mRNA and secretory levels of hBD-1 and -3 were down-regulated by CSE, while the mRNA and secretory level of hBD-2 were up-regulated by CSE. The iE-DAP or BAY 11-7082 treatment reversed the regulatory effects of CSE on levels of hBDs. Conclusion: The present study indicated that cigarette smoke could potentially modulate the expression of crucial molecules of NOD1 signal pathway and hBDs in human oral mucosal epithelium. NOD1 signal pathway could play an important role in the regulatory effects of CSE on hBDs levels in oral mucosal epithelial cells.

GLUT-1 immunoexpression in oral epithelial dysplasia, oral squamous cell carcinoma, and verrucous carcinoma.

Glucose transporters, such as GLUT-1, mediate the important mechanisms involved in cellular glucose influx, allowing cells to proliferate and survive. The significance of GLUT-1 expression in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) has been less explored, and no study has investigated it in relation to verrucous carcinoma (VC). We evaluated 30 cases each of OED, OSCC, and VC, graded further on the basis of their differentiation, immunohistochemically for GLUT-1 expression, along with 10 specimens of normal oral mucosa (NOM) as controls. In OSCC, GLUT-1 expression increased with the degree of dysplasia and increasing grade (P < 0.001). The expression in VC was predominantly membranous and intense, resembling well differentiated OSCC. This increase of GLUT-1 expression in OSCC along with the degree of dysplasia and the histologic grade reflects the expanding glycolytic response to hypoxia. This is the first study to have revealed prominent GLUT-1 expression in VC, highlighting its inherent metabolic capacity.