Expression of Vimentin and Ki-67 Proteins in Cervical Squamous Cell Carcinoma and their Relationships with Clinicopathological Features

Objectives: To investigate the expression of vimentin and Ki-67 proteins in cervical squamous cell carcinoma(CSCC) and their relationships with patient clinicopathological features. Materials and Methods:Fifty-sevenCSCC samples archived in Department of Pathology in the First Affiliated Hospital of Wenzhou MedicalUniversity were selected. The expression of vimentin and Ki-67 proteins in CSCC tissue were detected usingimmunohistochemical SP method, and correlations between them and their relationships with clinicopathologicalfeatures were analyzed. Results: Among 57 CSCC tissues, there were 43 with positive expression of Vimentin,and the positive rate was 75.4%; there were 57 cases with positive expression of Ki-67, and the positive ratecame up to 100.0%. The results of Pearson correlation analysis displayed that the expression of vimentin had asignificantly-positive correlation with Ki-67 in CSCC tissue (r=0.984, co0.000). The expression of both Ki-67 andvimentin was intimately associated with the presence or absence of local invasion and lymph node metastasisas well as differentiated degrees of the tumor (P=0.003, 0.017, 0.000; P=0.001, 0.008, 0.003) instead of the age,tumor size and clinical staging (P>0.05). Conclusions: Epithelial-mesenchymal transition (EMT) tends to appearin poorly-differentiated CSCC tissue, and the up-regulation of vimentin expression is accompanied by highexpression of Ki-67, suggesting that invasion and metastasis readily occur in these tumor cells.     

EGFR Inhibitor Enhances Cisplatin Sensitivity of Oral Squamous Cell Carcinoma Cell Lines

Epidermal growth factor receptor (EGFR) is involved in multiple aspects of cancer cell biology. EGFR has already been identified as an important target for cancer therapy, with various kinds of EGFR inhibitors currently used in treatment of several human cancers. Recently, EGFR and its downstream signaling pathways were identified as being associated with cisplatin sensitivity. In addition, EGFR inhibitors have shown significant promise for patients who failed cisplatin-based therapy. In this study, we investigated whether treatment with an EGFR inhibitor improves cisplatin sensitivity in oral squamous cell carcinoma (OSCC) cell lines. The effects of a combination of AG1478, a specific EGFR tyrosine kinase inhibitor, with cisplatin were evaluated in cultured OSCC cell lines and cisplatin-resistant sublines. Higher expression of EGFR and p-EGFR was found in the two cisplatin-resistant cell lines compared with the corresponding parental cell lines. In addition, augmented inhibition of OSCC cell growth by the combination of AG1478 with cisplatin was found in both cell lines. These results suggest that the combination of an EGFR inhibitor and cisplatin may be useful as a rational strategy for the treatment of patients with oral cancer with acquired cisplatin resistance.     

Circulating MicroRNA-21 Expression as a Novel Serum Biomarker for Oral Sub-Mucous Fibrosis and Oral Squamous Cell Carcinoma

Background: Circulating miRNAs (miRs) in the biofluids such as serum and plasma act as potential biomarkersfor early diagnosis, treatment and prognosis. In the present study, an attempt made to see the expression of miR-21in serum of 20 cases of Oral sub-mucous fibrosis (OSMF), 20 cases of Oral squamous cell carcinoma and 40 healthyvolunteers. The expression of miR-21 was evaluated in relation to different demographical and clinicopathologicalfeatures such as sex, tobacco, pan-masala, alcohol, smoking and clinical staging respectively with an aim to identifycorrelation with oral pre-cancer and cancer stages. Materials and Methods: The relative expression level of miR-21was determined by quantitative real-time RT-PCR (qRT-PCR) in the sera of 20 OSCC, 20 OSMF patients and 40healthy subjects as a control. Association between expression of miR-21 and OSCC clinical stages and demographicalparameters such as sex, pan-masala, tobacco, smoking, alcohol have also been analyzed in detail. Results: The resultsobtained by t-test revealed significant increase in the expression level of miR-21 in OSCC as compared to OSMF. Thestudy also revealed the positive correlation between higher miR-21 expression and pan-masala chewers as shown byt-test. The statistical test, ANOVA has also indicated a positive correlation between up-regulation of miR-21 in theclinical stages of the OSCC. Conclusion: The results of present study indicated up-regulation of circulating miR-21 inserum of OSCC as compared to OSMF (p=0.001), this study also elucidated the positive correlation between miR-21expression in OSCC/OSMF patients, only one demographical parameter (Pan-masala) and negative correlation forother parameters such as sex, tobacco, smoking, alcohol etc. Other findings suggested a significant increase (p=0.000)in the expression of miR-21 in clinical staging (I-IV) of oral cancer. More studies are needed to validate it as potentialdiagnostic and prognostic biomarker for OSMF and OSCC for better management.     

Comparison of PD-L1 Expression in Oral Squamous Cell Carcinoma and Premalignant Lesions of Oral Cavity

Objective: Objectives of this study were to compare  expression of Programmed Death-Ligand 1(PD-L1) protein in oral squamous cell carcinoma (OSCC) and oral potentially malignant disorder (OPMD) cases; and to compare the PD-L1 protein expression in histological grades of OSCC and also in OPMD’s with Dysplasia and without Dysplasia. Materials & Methods: In this study,  25 cases of Oral squamous cell carcinoms, 25 cases of Oral Potentially Malignant Disorders and 10 cases of non-neoplastic oral mucosa (control) cases were included. FFPE blocks of OSCC and OPMD cases were contributed by Department of Pathology, Histopathology Division,Pakistan Institute of Medical Sciences, Islamabad. Immunohistochemical staining of cases with PD-L1 monoclonal antibody (1:100; Dako) was carried out at Histopathology division , PMC Labs,  Peshawar Medical College,Peshawar, Riphah International University, Islamabad . Epithelial cells (membranous and cytoplasmic) positivity was observed for PD-L1 Antibody. Data was analyzed in SPSS version20. For qualitative variables frequencies and percentages were calculated whereas for quantitative variables means and standard deviations were recorded. The Chi-square test was applied to evaluate the significant difference in categorical variables . p-value of ≤0.05 was taken as significant. Results: PD-L1 expression in OSCC cases turned out to be 48% (n=12/25) as compared to 8% of OPMD cases (n=2/25) with a significant p value of 0.002 and all non-neoplastic oral mucosa cases were negative. PD-L1 expression in high grade OSCC cases was quite high (61% n=11/18) as compared to low grade OSCC (14% n=1/7) cases with a significant p value of 0.035. Conclusion: A statistically significant increased PD-L1 expression was noted in  OSCC as compared to OPMD. Expression of PD-L1 was more intense in high grade OSCC cases. The relation of PD-L1 expression to age ,gender or location of OSCC and OPMD cases , and presence of dysplasia in OPMD cases was statistically not significant.     

The Occurrence of Activated Leukocyte Cell Adhesion Molecule (ALCAM) and Its Predictive Factors in Patients with Oral Squamous Cell Carcinoma

Objectives: To determine the occurrence of Activated Leukocyte Cell Adhesion Molecule (ALCAM) and its predictive factors in patients with oral squamous cell carcinoma (OSCC). Methods: This cross sectional study was concocted on 102 patients with OSCC referred to Imam Khomeini Hospital of Tehran during 1997-2015. The data collection tool a checklist consisted of demographic and pathologic (lymph node involvement, differentiation, tumor size and tumor location) characteristics which extracted from patients’ medical records. To evaluate ALCAM, a new sample of tumor tissue was prepared from archive. Finally, the multivariable logistic regression model was used to determine the predictive factors of ALCAM by STATA14. Results: the number (%) of men and women were 70 (68.6) and 32 (31.4%), respectively. The mean age (S.D) of participants was 61.7 (15.6) years. Of the total samples, 32 (38.2), 19 (18.6), 36 (35.3) and 8 (7.8%) samples were related to the tongue, oral mucosa, skin and lips, respectively. More than half of the tumors had good differentiation and lymph node involvement and 74.5% were ≥20 mm. Also, 79.41% of the samples were positive for the overall incidence of ALCAM. The most important predictors of the overall incidence of ALCAM were tumor size (OR: 3.46, 95% CI: 1.71 - 7.01) and tumor location (OR: 3, 95% CI: 1.03 - 8.72). Similarly, for incidence of cytoplasmic ALCAM were age (OR: 2.56, 95% CI: 1.38 - 4.76) and location of the tumor (OR: 3.23, 95% CI: 1.08 - 9.64). However, the only predictor of membranous ALCAM incidence was lymph node involvement (OR: 0.36, 95% CI: 0.19 - 0.66). Conclusion: The results of our study suggest preliminary evidence for the potential clinical application of ALCAM as a prognostic biomarker for OSCC which may be the basis for future clinical application, however further studies are recommended.     

Promoter Hypermethylation of LATS2 Gene in Oral Squamous Cell Carcinoma (OSCC) Among North Indian Population.

ABSTRACTLarge Tumor Suppressor (LATS2) gene are Tumor Suppressor gene, linked with epigenetic modifications. LATS2 promoter hypermethylation is an important epigenetic silencing mechanism leading to cancer. Cancer is the most common, vicious and dangerously increasing diseases of the world today, associated with high morbidity and mortality. Oral cancers (OC) are the blazing universal dilemma and is the sixth most frequent cancer observed in Indian population. Tobacco consumption is the main cause of the increase in OSCC. The association between LATS2 in the pathogenesis of cancers propose that their combination might be studied as a possible molecular marker for particular subgroups of patients. Therefore, the present study tried to investigate whether LATS2 promoter methylation was associated with oral squamous cell carcinoma (OSCC) in North Indian subjects. DNA methylation quantitative studies of LATS2 Tumor Suppressor genes were performed by methylation-specific polymerase chain reaction (MSP). 38 out of 70 patients (55 %) were found to be methylated for LATS2 gene, a statistically significant result was obtained (p-value < 0.005) for LATS2 genes. The results suggest that epigenetic changes may be related to the down-regulation of LATS2 expression. It can be concluded that LATS2 gene plays a significant role in the diagnosis of cancer and provide a better alternative as a diagnostic biomarker. Our data infer that a low LATS2 expression due to methylation may contribute to the cancer progression and could be useful for the diagnosis of OSCC. Therefore, investigation of promoter methylation in such genes may provide a biomarker which may prove to be useful in early detection of Oral Cancer.Keywords: Oral squamous cell carcinoma (OSCC), Epigenetic changes, LATS2 gene, Promoter Hypermethylation, Methylation-Specific PCR (MSP), BiomarkersAbbreviations: OSCC- Oral Squamous Cell Carcinoma; DNA- Deoxyribonucleic acid; LATS-Large Tumor Suppressor (gene); MSP-Methylation-Specific Polymerase Chain Reaction     

Expression of Laminin in Oral Squamous Cell Carcinomas

Background and objectives: Laminin is a significant basement membrane (BM) glycoprotein, the expression ofwhich reflects BM integrity more precisely than do other ECM proteins. The present study aimed to evaluate lamininexpression in oral squamous cell carcinomas OSCC and to determine any associations with clinico-pathologicalparameters (surgical margin status, lymph node involvement, survival and recurrence). Methods: Laminin expression wasevaluated in 31 cases of biopsy-proven OSCC by immunohistochemical staining and its association with prognosticatorsand the Brynes grading system was determined by appropriate statistical analysis. Results: We observed a significantincrease in linear staining pattern (p<0.001) at the tumour-host interface in well-differentiated OSCC cases, in contrastto poorly differentiated lesions which exhibited intense cytoplasmic expression within tumour cells. Higher cytoplasmiclaminin expression was seen in 33.3% of cases with involved surgical margins and 69.2% of cases with lymph nodemetastasis (along with weak/absent staining of laminin around the tumour-host interface – Basement membrane aroundtumour islands). Similarly, in 60% of the cases who died and in 81.8% of cases with tumour recurrence, moderatelyintense cytoplasmic laminin expression was seen within tumour cells. On comparing variables of the Brynes gradingsystem, significant cytoplasmic expression of laminin was linked with mild inflammation (p<0.0016) and increasedmitotic activity (p<0.008). Conclusion: Based on these observations, immunohistochemical expression of lamininmight be useful to evaluate histological differentiation and aggressiveness of OSCCs.     

Quercetin Induces Cell Cycle Arrest and Apoptosis in YD10B and YD38 Oral Squamous Cell Carcinoma Cells

Objective: Oral squamous cell carcinoma (OSCC) exhibits the highest lethality among head and neck cancers. Treatment for OSCC is limited due to diverse side effects. Quercetin is a natural flavonoid compound found in many kinds of plants and foods. Quercetin has been reported to be a modulator of proliferation and survival in various types of cancers due to its cytotoxic effects. We aimed to investigating chemopreventative roles of quercetin in YD10B and YD38 OSCC cells. Methods: For our study, two different types of OSCC cells were used. YD10B cells are tongue SCC cells with the p53 mutation and YD38 cells are lower gingiva SCC cells without the p53 mutation, respectively. The anticancer effects of quercetin were examined by cell viability, cell cycle, annexin-PI staining, and western blot. Result: Our results showed that quercetin decreased cell viability and induced G1 cell cycle arrest in YD10B and YD38 OSCC cells. Moreover, quercetin remarkably decreased the expression of cell cycle upregulating proteins and increased the expression of a CDK inhibitor. Quercetin also significantly increased the number of annexin-V-positive cells in a dose-dependent manner in both types of OSCC cells. This apoptotic potential of quercetin triggered cleavage of PARP followed by activation of p38 MAPK signaling pathway. Conclusion: In conclusion, this study demonstrates that quercetin shows different anti-cancer responses in OSCC with and without p53 mutation, respectively. Despite different p53 status in OSCC cells, quercetin led to apoptotic signals in both cells. Quercetin repressed cell proliferation with G1 cell cycle arrest and apoptosis by activating the p38 signaling pathway in two OSCC cells with different p53 status. These findings might provide new strategy for OSCC therapy by quercetin.     

口腔鳞癌患者外周血及癌组织中IL-17和Foxp3的表达及意义

目的 探讨口腔鳞癌患者外周血及癌组织中IL-17和Foxp3的表达及意义。方法 选取40例原发初诊经病理证实的口腔鳞状细胞癌患者为实验对象,术前流式细胞术测定外周血中IL-17与Foxp3所占比例及CD3、CD4、CD8、CD56的表达。组织标本为术中立即取材,SP法测定IL-17及Foxp3表达。选取20例良性肿瘤患者瘤旁正常口腔黏膜作组织对照。实验分为A（Ⅰ、Ⅱ期鳞癌患者）、B（Ⅲ、Ⅳ期鳞癌患者）、C（健康者）三组进行组间比较。 结果 A、B组外周血CD4+IL-17+ 细胞、CD4+Foxp3+细胞所占比例均显著高于C组, IL-17+、Foxp3+的表达水平呈正相关（r=0.772,P<0.05）,其中B组CD4+IL-17+、CD4+Foxp3+细胞所占比例高于A组（P<0.05）。与C组相比,A、B组外周血中CD3+和CD4+细胞的百分率、CD4+/CD8+的比值降低,CD8+细胞的百分率升高,差异均有统计学意义（P<0.05）。 IL-17、Foxp3在A、B组中阳性率均明显高于C组,在A组中阳性表达率低于B组,差异均有统计学意义（P<0.05）,且A、B组癌组织中IL-17与Foxp3的表达水平呈正相关（r=0.386,P<0.05）。结论 IL-17及Foxp3在口腔鳞癌的发生发展中起着一定的促进作用,这种作用可能与机体免疫状态有关。     

Oral Psychosomatic Disorders in Family Caregivers of Oral Squamous Cell Carcinoma Patients

Objective: To investigate the oral psychosomatic disorders (PSDs) in family caregivers (FCs) of oral cancer (OC) patients and to evaluate the correlation between these oral PSDs to severity of depression anxiety and stress. Methods: A total of 50 participants were included each in first degree relative (FDR), second degree relative (SDR) and control group. All the participants completed DASS-21 questionnaire and were subjected to thorough clinical history and oral examination. Results: All the FCs reported statistically significant higher mean levels of depression, anxiety and stress compared to controls (p˂0.001). A significantly greater number of FCs (40.00%) reported oral PSDs than control group (12.00%). Most prevalent oral PSD in FCs was aphthous stomatitis followed by oral lichen planus, bruxism, burning mouth syndrome and myofascial pain dysfunction syndrome. Moreover, there was a preponderance of these diseases in FDR (60.86%) compared to SDR (26.08%). FCs with moderate to very severe depression, anxiety and stress showed higher prevalence of these oral PSDs compared to the ones with mild depression, anxiety and stress. Conclusion: The observations of higher prevalence of oral PSDs in FCs with psychological alterations can enhance healthcare professionals’ awareness to better understand FCs’ oral healthcare needs.     

Ki-67 Prognostic Value in Different Histological Grades of Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma

Introduction: Abnormal cell proliferation appears to be a possible predictor of tumorigenesis, Ki-67 proteinexpression is closely related to the cell proliferation and could be used as a biomarker for the growth in the most ofhuman tumors. The aim of the study: Investigating of Ki-67 expression in the pathological grades of oral epithelialdysplasia and oral squamous cell carcinomas. Materials and Methods: The sample consisted of 30 formalin-fixed,paraffin-embedded specimens of oral epithelial dysplasia (OED), 30 other of oral squamous cell carcinomas (OSCC), and10 normal oral epithelium (NOE) were conventionally stained with hematoxylin and eosin and immunohistochemicallystained with Ki-67 monoclonal antibody. Results: Expression of Ki-67 was restricted to the basal layers in the normaloral epithelium whereas Ki-67 positive cells in oral epithelial dysplasia (OED) were located in the basal, suprabasaland spinous layers, Ki-67 expression was increased in high-risk cases. Ki-67 positive cells in well-differentiated(OSCC) were located mainly in the periphery of the tumor nests, in moderately-differentiated (OSCC) were locatedin both peripheral and part of a center of the tumor nests whereas it was diffused in most of the Poorly-differentiated(OSCC). Statistical analysis indicated a significant difference between the expression in (OED) and (NOE), (OSCC)and (NOE), and no differences between (OED) and (OSCC). Conclusion: This study has concluded that Ki-67 antigencould be used as a marker for the histological grading of OED and OSCC, Expression of Ki 67 increased according tothe severity of oral epithelial dysplasia.     

改良和传统肩胛舌骨上颈淋巴结清扫术对口腔鳞癌患者疗效及预后的影响

目的 分析改良和传统肩胛舌骨上颈淋巴结清扫术对口腔鳞癌(OSCC)患者的临床疗效和预后情况.方法 选取80例确诊为口腔鳞癌患者为研究对象,采用随机数字表法将所有患者随机分为对照组与试验组,每组40例,试验组给予改良的肩胛舌骨上颈淋巴结清扫术治疗,对照组采用传统的根治性颈淋巴结清扫术,比较两组患者手术时间、术中出血量、肩...     

Comparative Evaluation of SMAD-2 Expression in Oral Submucous Fibrosis and Reactive Oral Lesions

Background: The event of fibrosis encompasses involvement of definite immunological and molecular mechanisms. As quite a lot of pro-fibrotic pathways are concerned, a multipronged approach is obligatory to cognize the fibrotic events. SMAD signaling pathway hasn’t been studied oral fibrotic events.In the progression of cramming the SMAD signaling pathway in OSMF, the first initiator protein of the pathway was considered for evaluation in the present study. Materials and Methods: A total of 100 subjects consisting of 20 controls, 40 patients with reactive lesions such as Traumatic Fibroma, Epulis Fissuratum and Gingival Hyperplasia and 40 patients with Oral Submucous Fibrosis were recruited for the study. Tissue homogenates were assayed by quantitative sandwich enzyme immunoassay technique using Human Mothers Against Decapentaplegic Homolog 2 (Smad2). Results: SMAD 2 expression values showed significant difference between control and OSMF group. However, the difference between reactive lesions with control and OSMF were not statistically significant. Conclusion: Graded increase of SMAD 2 expression from control,reactive lesions and OSMF were observed accentuating the role of SMAD signalling pathway in fibro genesis. Further this can be validated to generate effective antifibrotic targets.     

Molecular Grading of Oral Squamous Cell Carcinomas Infected with EBV

Background: Squamous Cell Carcinoma (SCC) is a type of cancer that is often found in oral cavity and areas ofthe head and neck. Viruses are major etiological factors through production of factors that can disturb proliferationand apoptosis regulators such as p53i, c-myc and bcl-2. This study aimed to determine the molecular grading of oralsquamous cell carcinoma (OSCCs) infected with the Epstein-Barr Virus (EBV). Methods: Twenty-seven OSCC patientsunderwent biopsy to detect EBV infection through in situ hybridization for RNA EBV (EBER) and immunohistochemicalanalysis of latent membrane protein-1 (LMP-1) and EBV nuclear antigen-1 (EBNA-1). To assess molecular grades, cellproliferation and apoptosis regulator expression i.e. inactive p53 (p53i), c-myc and bcl-2, were immunohistochemicallyanalysed. Results: The cases were divided into two groups; infected and non-infected by EBV. Regression analysisshowed that only EBNA-1 expression could affect p53i expression. Based on regression equations molecular grading ofOSCCs infected by EBV was divided into three: Grade I (low), EBNA-1 expression was 7.60, and p53i expressionwas 9.74-17.5; Grade II (medium), EBNA-1 expression was 7.61-19.7, and p53i 17.5-30.1; Grade III (high), EBNA-1expression was 19.71, and p53i ≥ 30.1. Conclusion: In OSCC infected with EBV, only EBNA-1 expression caninfluence p53i expression.     

Correlation of Habits and Clinical Findings with Histopathological Diagnosis in Oral Submucosal Fibrosis Patients

Background: Oral submucosal fibrosis (OSMF) is one of the most prevalent premalignant conditions in Indiawhich is easy to diagnose but difficult to manage. At present it is considered as irreversible and incurable. It hasalso been referred to as an epidemic in India. Aims and Objectives: To correlate the frequency and durationof habits with clinical staging, functional staging and histopathological grading and to correlate the clinicaland functional staging with histopathological grading. Materials and Methods: The study included a total of90 subjects, 80 with OSMF in the experimental group and 10 patients in the control group. Patient personalhistory was recorded with chewing habits, including frequency and duration of chewing. The site of keepingthe quid, time duration and whether he/she swallows it or spits it were also noted. Clinical staging was doneon the presence of palpable fibrous bands. Functional staging was accomplished by measuring mouth opening.Incisional biopsy was done for all the patients for histopathological examination. Histopathological gradingwas according to Pindborg and Sirsat. Results: The experimental group comprised 71 males and 9 females, themajority of which were in the age group of 21-30 years. Correlation of habits with clinical staging, functionalstaging and histopathological grading were significant (p<0.05). Clinical and functional staging did not correlatewith histopathological grading, but the correlation of clinical and functional staging was highly significant(p<0.01). Conclusions: The widespread habit of chewing gutkha is a major risk factor for OSMF, especially inthe younger age group. In this study, it was found that with increase in the duration and frequency of the habitthe severity of the disease increased.     

Salivary MMP-9 as a Biomarker for the Diagnosis of Oral Potentially Malignant Disorders and Oral Squamous Cell Carcinoma

Objective: To compare the salivary MMP – 9 concentration among subjects with oral squamous cell carcinoma (OSCC), oral potentially malignant disorders (OPMD), tobacco users, and control groups. Materials and methods: A total of 88 subjects were enrolled and divided into four study groups viz., OSCC (n=24), OPMD (n=20), tobacco habits (n=22), and healthy controls (n=22). All subjects gave unstimulated saliva samples for the evaluation MMP – 9 by ELISA kit. Demographic information like age, gender, type of tobacco, and duration of the habit were recorded. Results: Subjects with OSCC and OPMD had significantly higher mean MMP-9 levels than subjects with tobacco habits and control groups (P<0.001). Also, poorly differentiated OSCC group had significantly higher mean saliva MMP-9 than moderate and well-differentiated OSCC. The optimal cut-off point was 214.37 ng/mL with a sensitivity of 100% and specificity of 59% for OSCC versus the control group. The optimal cut-off point was as 205.87 ng/mL with a sensitivity of 100% and a specificity of 54% for OPMD versus the control group. Conclusion: The data obtained from this study indicated that OSCC and OPMD had an increased level of salivary MMP-9. Salivary MMP-9 could be a useful, non-invasive adjunct technique in the diagnosis, treatment, and follow-up of OSCC and OPMD.     

Stathmin! An immunohistochemical analysis of the novel marker in Oral Squamous Cell Carcinoma and Oral Leukoplakia.

Background: Stathmin is an intracellular phosphoprotein that controls the microtubule dynamics by further regulating proper attachment and alignment of chromosomes in a dividing cell. Thus, any mutation or aberrantly expressed protein that reduces the fidelity of spindle assembly will enhance chromosomal instability contributing to aneuploidy. Oral Squamous Cell Carcinoma is an extensively studied malignancy that occurs due to accumulated genetic changes due to carcinogens. The current study is done to evaluate the stathmin role and its expression in OSCC and Oral epithelial dysplasia (OED). Objective: The aim of the present study is to evaluate the role of stathmin in OSCC and Oral dysplasia and also to correlate the expression of Stathmin with respect to the different histopathological grades of OED and OSCC. Materials and Methods: 30 neutral buffered formalin fixed, paraffin embedded (FFPE) tissues of Oral Leukoplakia/OED and 30 FFPE tissues of OSCC were subjected to immunohistochemistry with stathmin antibody. Five fields of each case with 300 cells were examined and a mean percentage of positive–stained slides were determined. The percentages were recorded accordingly with their respective histological grades. The results were analysed statistically. Results: The results of the present study demonstrated higher mean values of stathmin in tissues with OSCC (2.50) compared to leukoplakia (2.11) and normal tissues (0.00) with a high level of statistical significance (0.0001). There is also an increase in the percentage levels of stathmin with increase in the histological grade of differentiation in OSCC as well as leukoplakia. Conclusion: The present study found a statistical correlation between increased grades of the disease with expression levels of stathmin. This confirms that stathmin expression can contribute to disease progression and that stathmin might have a potential role as an early diagnostic biomarker and can be a therapeutic target for OSCC.     

Quantitative Proteomic Analysis of Non-Tobacco Associated Oral Squamous Cell Carcinoma Reveals Deregulation of Cytoskeletal and Apoptotic Proteins

Background: The exact etiology of non-tobacco associated oral squamous cell carcinoma (NT-OSCC) is still unknown. The lack of established biomarkers for oral NT-OSCC has resulted in less effective management and poor prognosis. Here, we report for the first time a panel of potential markers identified from the quantitative proteomic analysis of NT-OSCC using two-dimensional gel-electrophoresis (2D-GE) using matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF) coupled with mass spectrometry (MS) and further analysis using protein analysis through evolutionary relationships (PANTHER) database. Objective: To quantitatively analyze the proteomic profile of non-tobacco associated oral squamous cell carcinoma. Methods: Twenty fresh tissue samples were collected from healthy controls and NT-OSCC, ten each, and were subjected to proteomic analysis. Sample quantification for the presence of protein was done using Bradford assay and bovine serum albumin was used as a standard protein to obtain the standard graph. Fractionation of protein was done using sodium dodecyl sulphate – polyacrylamide gel electrophoresis (SDS-PAGE) and they were separated based on their molecular weight. MS analysis was done and the purified peptides were analysed using MALDI-TOF. PANTHER database for functional classification and pathway analysis was done for identification of protein expression. Results: Our approach of combining 2D-GE with MS identified four candidate proteins including keratin, alpha-1-antitrypsin (AAT), S100 and serpin B5 with significant differential expression in NT-OSCC as compared with healthy controls. The results showed that the levels of these proteins were significantly upregulated in NT-OSCC when compared to the healthy controls that suggests that these proteins can be used as candidate targets for NT-OSCC therapeutics. Conclusion: The differentially expressed proteins are found to be involved in apoptotic signalling pathways, cytoskeletal dynamics and are known to play a critical role in oral tumorigenesis. Put together, the results provide available baseline information for understanding the development and progression of NT-OSCC. These identified proteins on further validation may be used as potential biomarkers in future for early detection and predicting therapeutic outcome of patients with NT-OSCC.