Well-differentiated Oral Squamous Cell Carcinoma Research Articles

Stromal myofibroblasts in potentially malignant and malignant lesions of the oral cavity.

Previous studies have demonstrated that myofibroblasts in the adjacent stroma are involved in the development and progression of malignant tumors. The aim of this study was to investigate the involvement of myofibroblasts in the progression of oral squamous cell carcinomas (OSCCs) by determining myofibroblast density in potentially malignant and malignant oral lesions. A total of 69 potentially malignant oral lesions (leukoplakias with mild, moderate or severe dysplasia), 90 OSCCs (well-, moderately and poorly differentiated), eight oral verrucous carcinomas and 29 fibrous hyperplasias were examined for the presence of myofibroblasts using immunohistochemical detection of isoform α of smooth muscle actin. Myofibroblasts were not identified in the adjacent stroma of fibrous hyperplasias and potentially malignant oral lesions, whereas 59.8% of the oral carcinomas exhibited myofibroblasts in various densities. The density was significantly higher in moderately and poorly differentiated OSCCs when compared with well-differentiated tumors (P=0.04 and P=0.007, respectively). In verrucous carcinomas, the specific variant of well-differentiated OSCC, stromal myofibroblasts were not detected. The results of the present study demonstrated that immunodetection of myofibroblasts does not aid with the determination of the malignant transformation potential of oral dysplasias, although moderately and poorly differentiated tumors exhibited a significantly higher density of myofibroblasts. The results reinforce the hypothesis that myofibroblasts may contribute to oral tumorigenesis, indicating that verification and monitoring of such may serve as a putative marker of OSCC behavior.

MutSα and MutLα immunoexpression analysis in diagnostic grading of oral epithelial dysplasia and squamous cell carcinoma.

This study explored the expression of DNA mismatch repair (MMR) proteins in a range of oral biopsies. We further evaluated the significance of MMR protein expression combined with basic demographic data in differentiating grades of oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC). Immunohistochemical expression of MutSα (hMLH1 and hPMS2) and MutLα (hMSH2 and hMSH6) were compared in 98 formalin-fixed paraffin embedded oral biopsies: 21 normal, 24 mild-dysplasia (MD), 8 moderate-to-severe-dysplasia (SD), and 45 OSCC. Expression of hMLH1, hPMS2, and hMSH2 was reduced in MD, SD, and OSCC compared with the normal. Reduced hMSH2 immunoreactivity discriminated poorly differentiated OSCC from well-differentiated OSCC. The diagnostic model correctly classified 71.4% of cases and revealed that hPMS2-negative biopsies were more likely to be cancerous (odds ratio [OR], 0.11; 95% confidence interval [CI], 0.000-0.813; P= .040). The results suggested a diagnostic role for MMR proteins in OED and OSCC.

MCP-1 as a potential target to inhibit the bone invasion by oral squamous cell carcinoma.

Bone invasion is a common complication of oral squamous cell carcinoma (OSCC), and this study sought to explore whether suppressed expression of monocyte chemotactic protein-1 (MCP-1) can be used to inhibit the bone invasion by OSCC. Strong staining of MCP-1 protein was observed from 10 archival blocks of OSCC by immunohistochemistry (IHC). Real-time PCR showed MCP-1 mRNA was highly expressed by OSCC cell lines (SCC25, HN5, and Tca8113), and SCC25 cells had the highest expression. An expression construct of a dominant negative variant of MCP-1 with 7 amino acids truncated (7ND), in the vector pcDNA was used to transfect SCC25 cells, and resultant stabilized SCC25 cells (SCC25-7ND) were generated by antibiotic selection. 10% conditioned media (CM, supernatant) of SCC25-7ND cells efficiently inhibited the formation of human osteoclasts grown from CD14(+) monocyte subpopulation, comparing with 10% CM of SCC25 cells. Further, cells of SCC25 or SCC25-7ND were injected onto the surface of calvariae of nude mice to establish an animal model of bone invasion by OSCC. H&E staining showed well-differentiated OSCC was formed in both groups, tumour cells invading the bone while osteoclasts locating in typical resorption lacunae. TRAP staining indicated significantly fewer osteoclasts were found in calvariae with cells of SCC25-7ND in comparison to cells of SCC25. These data demonstrate the relevance of MCP-1 with research on bone invasion by OSCC, and suggest the potential value of MCP-1 as a target to inhibit this common complication.

Expression of Mucin 4 in leukoplakia and oral squamous cell carcinoma: An immunohistochemical study.

Background:Mucins are large glycosylated proteins that act as a selective molecular barrier on the epithelial surface and engage themselves in morphogenetic signal transduction pathways. MUC4 (Mucin 4) is a transmembrane mucin, that protects and lubricates the mucous membranes of the human body and involves itself in various cellular functions like growth, differentiation and signaling. An aberrant expression of MUC4 has been demonstrated in various human cancers. A thorough literature survey shows very few studies about MUC4 expression in normal and cancerous oral mucosa.Aim:Our study aimed at investigating the expression pattern of MUC4 in normal oral mucosa, oral leukoplakia and oral squamous cell carcinoma (OSCC) in an attempt to analyze its role played in oral carcinogenesis.Materials and Methods:Formalin-fixed paraffin-embedded tissues of five cases of normal tissue, 15 cases of leukoplakia, 10 cases of well-differentiated squamous cell carcinoma and 10 cases of moderately differentiated squamous cell carcinoma were retrieved from the archives of the department and MUC4 antigen was immunohistochemically localized.Statistical Analysis:The result was subjected to statistical analysis using Pearson's Chi-square test and an intergroup analysis was performed using one-way analysis of (ANOVA).Results:A total of 46.7% of leukoplakia and 70% of OSCC were stained positive with MUC4 antigen. Maximum intensity of staining was noted in well-differentiated OSCC. A steady increase in MUC4 staining was noted from normal oral tissues to leukoplakia to OSCC.Conclusion:The findings of the study suggest that MUC 4 plays a vital role in the pathogenesis of OSCC and can be regarded as a useful marker for oral dysplasia and OSCC.

Oral squamous cell carcinoma with pseudo-glandular differentiation

Conventional oral squamous cell carcinomas are readily recognized histopathologically but the presence of additional atypical features may be challenging from a diagnostic point of view. We present a case of a well-differentiated oral squamous cell carcinoma with pseudo-glandular differentiation and discuss the possible differential diagnoses on a histopathological basis. Accurate diagnosis is imperative for timely and appropriate intervention and denotes distinctive prognostic implications. The presence of perivascular and perineural infiltrations as observed in this case would indicate the need for further post-operative therapeutic decision-making aimed at controlling local spread as well as distant metastases.

Suitability/Unsuitability of Cell Proliferation as an Indicator of Malignant Potential in Oral Lichen Planus: an Immunohistochemical Study

Oral lichen planus (OLP) is categorized as premalignant lesion although its malignant potential is a matter of controversy. The objective of this study was to investigate Ki67 expression in OLP, oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). Expression of Ki67 was evaluated through immunohistochemistry (IHC) in groups of A (17 cases of epithelial hyperplasia), B (16 cases of OLP), C1 (10 cases of mild epithelial dysplasia), C2 (10 cases of severe epithelial dysplasia), D1 (10 cases of well-differentiated OSCC), and D2 (10 cases of poorly-differentiated OSCC). There was a significant difference in Ki67 expression based on quantitative analysis among the six studied groups as well as group B compared bilaterally with groups C2, D1 and D2 (p< 0.0001). However, there was no significant difference between groups B and C1 or between groups D1 and D2 (p> 0.05). Based on the results of the present study it may not be possible to definitely consider malignant transformation potential for OLP. However, expression of Ki67 was significantly higher in OLP than that of epithelial hyperplasia with no significant difference from that of mild epithelial dysplasia. This should be considered by clinicians to carefully and regularly follow up OLP lesions to detect potential subtle changes at an early stage.

NANOG expression correlates with differentiation, metastasis and resistance to preoperative adjuvant therapy in oral squamous cell carcinoma

NANOG protein, a transcription factor expressed in embryonic stem cells, is overexpressed in tumor development. Although studies investigating the function of NANOG in cancer have shown that it plays several roles, such as in cell proliferation, invasion and metastasis, the overall function of NANOG in cancer cells has remained elusive. In the present study, NANOG expression in oral squamous cell carcinoma (OSCC) was examined to determine its potential clinical significance. The expression of NANOG protein was assessed in 60 patients with OSCC by immunohistochemistry, and its correlation with clinicopathological factors and metastasis was evaluated. NANOG protein levels in human OSCC cell lines were determined by western blotting and immunofluorescence staining. NANOG protein expression was identified in 52 cases (86.7%) and expression levels were higher in primary foci of poorly differentiated OSCC than in those of well-differentiated OSCC, indicating that NANOG expression is associated with OSCC differentiation. Regardless of the differentiation levels of primary foci, NANOG expression levels in metastatic foci were extremely high. In addition, NANOG expression in metastatic foci was maintained at high levels following preoperative adjuvant therapy. Furthermore, NANOG protein was detected at an identical level in human OSCC cell lines. These data indicate that NANOG-expressing OSCC cells tend to metastasize and that metastatic tumors expressing NANOG may be resistant to preoperative adjuvant therapy, including chemoradiation. Thus, assessment of NANOG expression may assist the strategy for treatment of OSCC metastasis.

Simultaneous quantification of nucleoproteins and comparison of methyl green-pyronin Y and Feulgen staining in sections of oral squamous cell carcinoma, dysplastic lesions and normal mucosa

A fundamental difference between normal cells and tumor cells is the proliferative activity of the nucleus and nucleolus, which increases progressively from normal to oral dysplastic mucosa to oral squamous cell carcinoma (OSCC). This activity is evaluated routinely using hematoxylin and eosin (H & E) staining, but in some cases, inter-observer variability occurs among pathologists. We evaluated cellular proliferation by staining sections with the methyl green-pyronin Y procedure and the Feulgen reaction. We also compared the efficacy of methyl green-pyronin Y and Feulgen staining for studying nuclear and nucleolar features in oral dysplastic mucosa and in different grades of OSCC. Sections cut from formalin fixed, paraffin embedded blocks of five normal mucosa, 15 dysplastic mucosa, 10 well-differentiated OSCC, 10 moderately differentiated OSCC and five poorly differentiated OSCC cases were stained with Hematoxylin and Eosin, methyl green-pyronin Y and the Feulgen reaction. The mean diameters of the nuclei and number of nucleoli showed significant differences. A progressive increase in diameter of the nucleus and number of nucleoli was observed from normal mucosa through poorly differentiated OSCC. We observed that methyl green-pyronin Y stain is more useful than Feulgen and hematoxylin and eosin for simultaneous quantitative assessment of both RNA and DNA. The simplicity of this technique makes it a valuable tool even for daily routine examination.

Immunohistochemical expression of endothelin protein in oral squamous cell carcinoma

Endothelin-1 (ET-1) plays a significant role in cancer biology and is considered as a potential target for molecular therapy. Alteration of ET-1 was detected in oral squamous cell carcinoma (OSCC). To evaluate the immunohistochemical expression and distribution of ET-1 in OSCC and to correlate its expression with the degree of histopathologic differentiation as well as the regional metastatic status of OSCC. The immunohistochemical expression and distribution of ET-1 was evaluated in total number of 30 cases of OSCC. The specimens were retrieved as formalin-fixed, paraffin-embedded blocks from the archival files. Detection of ET-1 expression in different grades of metastatic and non-metastatic OSCC was quantified using an image analyzer computer system. Statistical software package SPSS v. 19. ET-1 immunoreactivity was observed in all studied specimens. On comparing optical density values for ET-1 expression in different grades of OSCC, poorly differentiated OSCC showed a significantly greater expression than moderately differentiated OSCC, which was also significantly greater than well-differentiated OSCC ( P = 0.001). Moreover, ET-1 expression was higher in metastatic OSCC than that in non-metastatic OSCC ( P = 0.018). Increased ET-1 expression could enhance the aggressive behavior of poorly differentiated OSCC, especially metastasis. Accordingly, ET-1 could be a therapeutic target in OSCC.

The role of mast cells and angiogenesis in well-differentiated oral squamous cell carcinoma

Neo-vascularization is vital for an expansion and metastasis of a tumor and is influenced by a number of mediators. Mast cells are believed to release many pro-angiogenic mediators that may help in tumor expansion and invasion. This study describes the role of mast cells and angiogenesis in oral squamous cell carcinoma (OSCC). It was a prospective study in which 37 biopsies of well-differentiated OSCC were obtained. Micro-vessels were stained with cluster of differentiation (CD)-34 and mast cells were counted using the Toluidine blue stain. When compared to normal oral mucosal tissue, it was seen that micro-vessel density and mast cell density indeed increases significantly in squamous cell carcinoma; however, they are not correlated to each other. This study reports that angiogenesis does increase in OSCC and mast cells also invade the peri-tumor tissue, but they are not directly correlated.

Assessment of Cellular Proliferation in Oral Verrucous Carcinoma and Well-Differentiated Oral Squamous Cell Carcinoma Using Ki67: A Non-Reliable Factor for Differential Diagnosis?

Non-invasive oral verrucous carcinoma (OVC) and invasive well-differentiated oral squamous cell carcinoma (OSCC) have similar histopathologic findings but different biological behavior. These two malignancies must be correctly differentiated by pathologists. The aim of this study was to determine immunohistochemical (IHC) expression of Ki67 in OVC and well-differentiated OSCC. Expression of Ki67 was evaluated by IHC in 15 cases of epithelial hyperplasia with no dysplasia (A group), 15 cases of OVC (B group), 12 cases of microinvasive OSCC(C group) and 15 cases of well-differentiated OSCC (D group). There was a significant difference in Ki67 expression based on pattern distribution of immunostaining positive cells, with quantitative and semi-quantitative analyses, among the four groups ; also, between A group and each of the other three groups (P=0.0001). But there was no significant difference between B and C, C and D, and B and D groups (P>0.05). The three evaluation methods of Ki67 expression showed Ki67 (Mib-1) is not a good immunohistochemical marker to assess invasion status and differentiate OVC from well-differentiated OSCC; also, it cannot be used as a diagnostic tool to distinguish between variants of OSCC with similar grade.

Expression of basal cell keratin 15 and keratin 19 in oral squamous neoplasms represents diverse pathophysiologies.

Human epithelium contains keratin, which is expressed during differentiation. Depending on the target cell type, different types of keratin are expressed, and their alterations seem to represent changes in cell properties. The basal cells of oral epithelium express keratin 5 (K5), K14, K15 and K19, but their alterations in tumors are unclear. To address this issue and to seek possible diagnostic application, we examined the expression of these keratins in oral squamous cell carcinoma (OSCC) and squamous intraepithelial neoplasm (SIN). cDNA microarray analysis of 43 OSCC revealed slight upregulation of KRT14, downregulation of KRT15 and KRT19, and unaltered KRT5 expression. There were great variations in KRT15 and KRT19 expression across each cancer. Well-differentiated OSCC tended to express more KRT15 and less KRT19 compared to moderately- or poorly-differentiated OSCC. KRT15 was positively correlated with differentiation-related keratin, KRT13. These observations were further investigated by immunohistochemical examination. K5 and K14 were ubiquitously expressed in all 50 OSCC and 50 SIN examined. K15 and K19 were generally downregulated, but were considerably retained in about half of the cases and showed diverse expression patterns. K15-positive cancers tended to show a well-differentiated phenotype, and K19-positive cancers tended to show more invasive tumor fronts. Most K19-positive cancers appeared to develop with little associating SIN. K19 was consistently downregulated in SIN, while K15 was downregulated mainly in high grade SIN. In summary, K15 and K19, unlike K5 or K14, are expressed variably in both SIN and OSCC, which reflects the differences in their pathogenesis and biological behaviors, suggesting their prospective applications as markers for subclassifying OSCC and SIN.

Overexpression of Smad proteins, especially Smad7, in oral epithelial dysplasias

Transforming growth factor β, via membrane-bound receptors and downstream Smad2-4, 7, can modulate tumorigenesis. Smad2 and Smad3 heterodimerize with Smad4, and the complex migrates to the nucleus to regulate the expression of target genes. Smad7 is a key negative regulator of this signaling pathway. This study aimed to examine Smad2-4, 7 expression and phosphorylated Smad2-3 (p-Smad2-3) in oral epithelial dysplasia and compared it with normal oral mucosa, hyperkeratosis/epithelial hyperplasia and squamous cell carcinoma (SCC). Immunohistochemical staining of Smad2-4, 7 and p-Smad2-3, was performed for 75 samples of human oral mucosa, including hyperkeratosis/epithelial hyperplasia (n = 20), mild epithelial dysplasia (n = 11), moderate to severe epithelial dysplasia (n = 11), and SCC (n = 43). Normal buccal mucosa samples (n = 9) were also included. A significant increase in Smad7 expression was observed in the ascending order of samples of normal oral mucosa, hyperkeratosis/epithelial hyperplasia/mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, and well-differentiated oral SCC/moderately to poorly differentiated oral SCC. Additionally, significant increases in Smad7 expression were noted as compared with expression of Smad2-4 and p-Smad2-3 in lesions of hyperkeratosis/epithelial hyperplasia, mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, well-differentiated oral SCC, and moderately to poorly differentiated oral SCC. Our results indicate that Smad proteins, particularly Smad7, in oral epithelial dysplasia and SCC could contribute to the attenuation of Smads anti-proliferative signaling in cancer development. Smad7 could be a marker for risk of malignant transformation of oral epithelial dysplasia.

Association of cytokeratin 17 expression with differentiation in oral squamous cell carcinoma

PurposeThe aim of this study was to confirm the expression profile of cytokeratin (CK)17 in comparison with that of CK13 in oral squamous cell carcinoma (OSCC) and leukoplakia and to clarify an association of CK17 with the OSCC differentiation.MaterialsThe expression of CK17 and CK13 was immunohistochemically examined in 105 patients with OSCC and 108 patients with leukoplakia. A correlation of CK expression with clinicopathological variables was carried out. The over-expression levels of CK17 mRNA were analyzed by real-time RT-PCR in 5 OSCC cell lines (HSC-2, HSC-3, SAS, SQUU-A, SQUU-B).ResultsCK17 and CK13 were detected in 101 (96.2 %) and three (2.9 %) of the 105 OSCCs, respectively. CK17 was significantly expressed in well-differentiated OSCC compared to moderately/poorly differentiated OSCC (p < 0.01). As detected in 19 of the 34 dysplastic leukoplakias (55.9 %) and 36 of the 74 hyperplastic leukoplakias (48.6 %), CK17 was significantly expressed in dysplastic leukoplakias (p < 0.01). As detected in 11 of the 34 dysplastic (32.4 %) and 52 of the 74 hyperplastic leukoplakias (70.3 %), CK13 was significantly expressed in hyperplastic leukoplakias (p < 0.01). The relative expression of CK17 mRNA in HSC-2 was significantly higher than in HSC-3 and SAS (p < 0.05). Moreover, the relative expression of CK17 mRNA in SQUU-A was significantly higher than in SQUU-B (p < 0.05).ConclusionCK17 expression could be associated with the differentiation and the malignancy of OSCC. A combination pattern of CK17/CK13 might be a suitable marker of malignant transformation.

Amino acid profile of saliva from patients with oral squamous cell carcinoma using high performance liquid chromatography

Oral cancer is the leading cause of death worldwide and it is the eighth most common cause of cancer death. Cancer cells utilize more glucose and amino acids than their benign counterparts. Diagnosis of disease via the analysis of saliva is potentially valuable, as the collection of fluid is associated with fewer compliance problems than the collection of blood. Hence, the present study was undertaken to evaluate the comprehensive amino acid profiling of saliva by high performance liquid chromatography (HPLC). The study group comprised 16 subjects, of whom eight were classified as having well-differentiated oral squamous (OSCC) cell carcinoma (Group I) and eight were classified as having moderately differentiated oral squamous cell carcinoma (Group II). Eight healthy individuals comprised the control group (Group III). The results showed increased salivary levels of all the amino acids in both groups of OSCC patients (Groups I and II) when compared with healthy controls (Group III). Hence, our study showed higher levels of all amino acids in the saliva of OSCC patients than in the saliva of healthy controls. The increased levels may serve as a "diagnostic and prognostic marker" for oral squamous cell carcinoma and for further detection of metastatic spread.

Vascular endothelial growth factor and ki-67 antigen expression in relation to age and gender in oral squamous cell carcinoma.

Background and aims Vascular endothelial growth factor (VEGF) and Ki-67 antigen are contributing factors in this process cell proliferation and new blood vessels formation in tumor progression. This study was conducted to examine the relationship between the expression of VEGF and Ki-67 and gender and age of patients with oral squamous cell carcinoma (OSCC). Materials and methods Twenty-three archival samples of well-differentiated OSCC were examined immunohisto-chemically and assessed by obtaining Total Score (TS = proportion score × staining index). For statistical analysis, t-test and Pearson’s correlation were employed. P≤0.05 was considered statistically significant. Results The differences in VEGF expression between males and females (P = 0.43) and different ages (P = 0.88) were not significant. The differences in Ki-67 expression was between males and females (P = 0.67) and different ages (P = 0.88) were also not significant. A positive correlation of VEGF and Ki-67 expression was observed in males and females in addi-tion to ≤ 60 years age group (r = 0.22, r = 0.008, and r = 0.58, respectively; P < 0.05). The expression of VEGF had a nega-tive relation to Ki-67 in > 60 years group (r = −0.48, P < 0.05). Conclusion The expression of VEGF and Ki-67 between males and females and different ages were not significant among oral squamous cell carcinoma cases evaluated.

Up-regulation of neutrophil gelatinase-associated lipocalin in oral squamous cell carcinoma: Relation to cell differentiation

Neutrophil gelatinase-associated lipocalin (NGAL, also known as lipocalin2, LCN2) is a secreted glycoprotein with increased expression in solid tumors. The expression and functions of NGAL in oral cancer, however, remain unclear. We investigated the expression of NGAL in oral cancer tissues and oral cancer cell lines. By immunohistochemical examinations, NGAL expression was strongly up-regulated in well-differentiated OSCC tissues and moderately to weakly up-regulated in moderately to poorly differentiated OSCC tissues. In contrast, NGAL expression was weak or very weak in normal mucosa and leukoplakia. By western blot analysis, NGAL expression levels positively correlated with cell morphology patterns and loss of E-cadherin. In addition, the enzymatic activity of the NGAL/MMP-9 complex significantly correlated with the results obtained by zymographic analysis. In conclusion, NGAL expression is high in well-differentiated cancer, suggesting that NGAL may be a useful diagnostic marker of tumor-cell differentiation.

Outcome analysis of patients with well-differentiated oral cavity squamous cell carcinoma

The prognosis of well-differentiated oral cavity squamous cell carcinoma (OSCC) is better than less-well-differentiated neoplasms. The aim of this retrospective study was to identify prognostic factors in patients with well-differentiated OSCC. The 5-year outcomes of 467 patients with well-differentiated OSCC who underwent radical surgery and neck dissection were analyzed. In the entire cohort, the presence of pathological node metastases (pN+ vs. pN0) was an independent predictor of 5-year outcomes. In pN0 patients, tumor depth (≥8 mm) was the only independently prognostic factor for 5-year survival rates on multivariable analysis (disease-free survival [DFS], P=0.001, hazard ratio [HR]=2.634, 95% confidence interval [95% CI]=1.496-4.636; disease-specific survival [DSS], P<0.001, HR=6.794, 95% CI=2.364-19.525). In pN+ patients, level IV/V neck nodal metastases (DFS, P<0.001, HR=47.483, 95% CI=8.942-252.122; DSS, P<0.001, HR=14.301, 95% CI=5.337-38.323), and ≥3 positive nodes (DFS, P=0.037, HR=2.107, 95% CI=1.047-4.242; DSS, P=0.044, HR=2.093, 95% CI=1.020-4.295) were independently associated with 5-year outcomes. Our results suggest that a tailored treatment approach in well-differentiated OSCC patients should take into account the presence of either pN0 or pN+ disease.

Tumour infiltrating CD25+ FoxP3+ regulatory T cells (Tregs) relate to tumour grade and stromal inflammation in oral squamous cell carcinoma

This research is aimed to quantitate and characterize the subtypes of tumour infiltrating lymphocytes (TILs), in particular the presence of FoxP3+ Tregs in different grades of oral squamous cell carcinoma using monospecific antibody staining of formalin-fixed, paraffin-embedded tissue sections. The correlation between tumour grade and the intensity of tumour infiltrating lymphocytes was tried to be tested, to assume a putative linkage between them. Thirty-four cases of histologically proven primary oral squamous cell carcinoma (OSCC) of different grades of differentiation were assorted to groups 1-3. Three-micron sections of tissue were cut and captured on electrically charged slides (Vision BioSystem, Mount Waverley, Australia) and stained using monospecific antibody against FoxP3+ Treg phenotype (dilution 1:40, Mouse monoclone No: 236A/E7, Ab 20034, IgG1; Abcam, Cambridge, UK). Automated protocols were employed for staining and scoring of staining intensity using Bond™ system (Vision BioSystem). Significant difference in staining intensities (Tregs) was noted among the histologic grades of tumour, where well-differentiated OSCC had significantly low expression of FoxP3+ Tregs in comparison with moderately and poorly differentiated tumours. A significant linear correlation was established between tumour grade and the intensity of TILs, where high grade tumours (poor differentiation) were more markedly infiltrated. There was also a significant positive correlation between FoxP3+ Tregs and TILs in cases studied. The correlation of these three variables noted in the study (FoxP3+ Tregs, tumour grade and TILs) and their significance in a meta-analysis may prove useful in targeting patients with high-risk neoplasms for more aggressive treatment protocols and management strategies.

Overexpression of COX-2 in oral squamous cell carcinoma patients undergoing chemoradiotherapy.

Aims and Objectives:To evaluate the prognostic significance of cyclooxygenase-2 (COX-2) overexpression in oral squamous cell carcinoma (OSCC) patients undergoing chemoradiation therapy. Purpose of the study was to determine whether COX-2 could be used as a diagnostic and prognostic index in OSCC.Materials and Methods:Forty-four patients of SCC were included in the present study and immunohistochemical examination was done for COX-2 expression. Negative and <5% COX-2 positivity were taken as negative expression and ≥5% COX-2 positivity as positive expression group. ≥30% COX-2 positivity was taken as overexpressed group and <30% COX-2 positivity was taken as underexpressed group. All the data were analyzed statistically.Results:COX-2 overexpression in OSCC was found in 15.90% cases. The proportion of COX-2 overexpression was higher in patients with large tumor size than in those with small tumor size. The proportions of COX-2 positive expression cases were higher with cervical lymph node metastasis. Negative COX-2 expression was higher in well-differentiated OSCC and positive expression was higher in moderately differentiated tumors. COX-2 underexpressed cases had better response to chemoradiation therapy as compared to cases with overexpressed COX-2.Conclusion:COX-2 expression in OSCC can be used as a prognostic marker. Studies with large sample size and long-term follow-up are required to find out the exact role and prognostic significance of COX-2 expression in OSCC.