Oral cancer is one of the most common cancer worldwide, ranking 6th in position. But India ranks 1st among males and 3rd among females.1 oral cancer accounts for 2% 4% of all cancer cases. In some regions, the prevalence of oral cancer is higher, reaching around 45% in India.2 Despite advancements in medical interventions, morbidity and mortality rates have remained relatively unchanged over the previous 30 years. Males had rates of 6.6 and 3.1/100,000 morbidity and mortality, respectively, while females have rates of 2.9 and 1.4/100,000. 3 Additionally, the incidence of OSCC is increasing among young white individuals age 18 to 44 years, particularly among white women. The percentage of 5-year survival for patients with OSCC varies from 40-50%.4 Regardless of the easy access of oral cavity for clinical examination, Oral malignancies is usually diagnosed in advanced stages. Oral cancer accounts for approx. 3% of all malignancies in total, It is an aggressive malignant neoplasm with high mortality and morbidity occurring in middle aged and older individuals with 6.5 yrs survival rate, which is about 55% despite therapeutic advances.5 Majority are associated with tobacco consumption and are preceded by precancerous lesions. OSCC is a multi-step process that involve genetic events leading to modification of normal function of oncogenes and tumour suppressor genes. Earliest morphological changes that could be detected are appearance of premalignant lesions that includes leucoplakia and erythroplakia.50% of leucoplakias exhibits dysplasia and overall malignant transformation potential of about 0.1-2%. 6 Loss of epithelial morphology and acquisition of mesenchymal characteristics termed as epithelial to mesenchymal transition, which is typical for carcinoma. Cells preceding EMT exhibits down regulation of many epithelial markers including E-Cadherin, 1 desmoplakin, cytokeratin, Claudins.7 Specific pattern of e- cadherin and p-53 expression can predict invasiveness and may be used as markers for early diagnosis. E cadherin plays a role in adhesion of cells. E-cadherin has a major role in establishing cell polarity and in maintaining normal tissue architecture. The intracellular domain of E-cadherin is linked to the actin cytoskeleton through its interaction with its cytoplasmic-binding partners, the catenin. 8 The loss of function of the p53 tumour suppressor gene is linked to nearly all human cancers, indicating the importance of p53 in cancer development. P53 is a tumour suppressor protein produced by the p53 gene.9 Loss of P53 function results in a loss of cell cycle control and the accumulation of damage-induced mutations, which leads to cell malignancy. P53 protein inactivation occurs when the p53 gene is mutated or when the P53 protein is stabilised by binding to other proteins such as MDM2. P53 protein expression has been studied extensively in oral cancers and pre-malignancies. In oral pre-malignancies, P53 protein expression ranges from 17 to 67 %, while in oral cancers, P53 protein expression ranges from 11 to 75 %.10 Combination of relevant data, adequate sampling, histo-pathological examination, immune-histo-chemistry are important to identify neoplastic lesions with high malignant potential of invasion and metastasis. Correlation of local invasiveness, metastatic potential of tumour and cellular proliferation can be done using e cadherin and p53 expression. Hence detection of high risk cases for distant metastasis at an early stage is mandatory for better prognosis. The present study was undertaken to systematically evaluate the immune-histo-chemical expression of E-cadherin and p53 in normal and neoplastic oral epithelium, and to compare their expression across different tumor grades, with the goal of determining whether these markers can independently or jointly serve as reliable predictors of the biological behavior of oral epithelial tumors.

The present study is the descriptive study conducted in the Department of Pathology, GANDHI HOSPITAL for a period of two years. Duration of Study: October 2019 to October 2021 Sample size: A total of 60 cases with normal and oral Squamous cell cancer cases were included in the study. Normal oral epithelium and all the cases of oral squamous cell cancer diagnosed on H&E were subjected to IHC staining specific for E-cadherin and p53. Inclusion Criteria: All properly labelled and fixed tumour specimen of patients of all ages and both sexes diagnosed as oral squamous cell cancer and normal oral epithelium on routine H&E are included in the study. Exclusion Criteria: • All cancers of oral cavity other than oral squamous cell cancer • Autolysed/necrosed tissue specimens • Inadequate biopsy. Investigations: • Surgical profile and CT/MRI of head and neck • Biopsy from suspicious area. • Routine H&E staining • IHC staining for E-cadherin and p53 expression • Evaluation of slides by experts 48. Statistical Analysis: The SPSS 22 software was used for statistical analysis. The data is presented in the form of percentages. The p-value of <0.05 was considered statistically significant.

TABLE-1: SOCIO-DEMOGRAPHY Age Group (in yrs) Total No. of patients Percentage 31-40 17 28.33% 41-50 16 26.66% 51-60 21 35.00% 61-70 5 8.33% >70 1 1.66% Gender Male 46 76.66% Female 14 23.33% Betelquid chewing 48 80% Alcoholic 54 90% Smooker Non-Chronic 50 98% Chronic 1 2% Oral hygiene Bad 46 76.66% Good 14 23.33% Table-2: Lesion features Site No of cases Percentage Lip 1 1.66% Buccal mucosa 18 30.00% Tongue 24 40.00% Floor of mouth 1 1.66% Retromolar trigone 3 5.00% Hardpalate 8 13.33% Softpalate, uvula 3 5.00% Anteriorpillar 2 3.33% Type of tumour Malignant 35 58.33% Benign 25 41.66% Table: 3- Association between E-Cadherin & p-53 with Gender Gender E- Cadherin P-53 Positive (%) Negative (%) Positive (%) Negative (%) Male 22(75.9) 4(66.7) 20(74.1) 6 (75) Female 7(24.1) 2 (33.3) 7 (25.9) 2(25) P-Value 0.638 0.958 Table-4: Distribution based on site of lesion malignant tumors with E-Cadherin &p-53 SITE E-Cadherin P-53 Positive Negative Combined (%) Positive Negative Combined (%) Lip 1 0 1(2.8) 0 1 1(2.8%) Buccal mucosa 8 2 10(28.6) 8 2 10(28.6%) Tongue 14 2 16(45.7) 12 4 16(45.7%) Floorof mouth 0 1 1(2.8) 1 0 1(2.8%) Hardpalate 5 0 5(14.3) 4 1 5 (14.3%) Softpalate, uvula 1 1 2(5.7) 2 0 2(5.7%) Table-5: Association between E-Cadherin & p-53 with tumour grade Tumour grade E-Cadherin P-53 Positive Negative Total (%) Positive Negative Total (%) Well Differentiated 20 (57.14%) 3 (8.57%) 23 (65.7%) 16 (45.71%) 7 (20%) 23 (65.7%) Moderately Differentiated 7 (20%) 2 (5.71%) 9 (25.7%) 9 (25.71%) 0 (0%) 9 (25.7%) Poorly Differentiated 2 (5.71%) 1 (2.85%) 03 (8.6%) 2 (5.71%) 1 (2.85%) 03 (8.6%) P-Value 0.0004* <0.001* *statistically significant Results: A total of 60 cases, comprising both normal subjects and patients with oral squamous cell carcinoma (OSCC), were included in this study. There was a marked male predominance, with males accounting for 77% of the cases, compared to 23% females, resulting in a male-to-female ratio of 3.28:1. The majority of patients (35%) fell within the 51–60 years age group, followed by 28% in the 31–40 years category, and 27% within 41–50 years. The remaining distribution included 8% between 61–70 years, and 1.66% older than 70 years. The mean age of patients was 49.81 ± 10.56 years. Among the 60 cases, 90% reported alcohol consumption, of whom half were chronic alcoholics. Regarding tobacco use, 85% were smokers, with 2% identified as chronic smokers. Poor oral hygiene was observed in 76.66% of cases. Analysis of lesion sites revealed that the tongue was the most common location (40%), followed by the buccal mucosa (30%), hard palate (13%), soft palate (5%), uvula (5%), retromolar trigone (5%), anterior pillar (3%), lips (2%), and the floor of the mouth (2%). E-cadherin was positive in 82.85% of cases and negative in 17.14%. Among positive cases, 76% were male and 24% female. Notably, E-cadherin expression was reduced in neoplastic oral epithelium compared to normal oral mucosa. Of the cases positive for E-cadherin (n=29), the most frequent lesion site was the tongue (14 cases), followed by buccal mucosa (8 cases), hard palate (5 cases), soft palate (1 case), and lower lip (1 case). Immunohistochemical analysis demonstrated p53 positivity in 77% of the cases, with 23% negative. Among p53-positive cases, 74% were male and 26% female. The distribution of lesions among p53-positive cases (n=27) was as follows: tongue (12 cases), buccal mucosa (8 cases), hard palate (4 cases), soft palate (2 cases), and floor of mouth (1 case). Regarding the histopathological grading, well-differentiated tumors constituted 65.71% of the cases, moderately differentiated tumors 25.71%, and poorly differentiated tumors 8.5%. Among the well-differentiated tumors, 57.14% were E-cadherin positive, while E-cadherin positivity was observed in 20% of moderately differentiated and 5.7% of poorly differentiated tumors. This association was statistically significant (Chi-square: 23.12, p=0.004). Additionally, p53 positivity was identified in 46% of well-differentiated, 26% of moderately differentiated, and 6% of poorly differentiated tumors, also representing a statistically significant correlation (Chi-square: 6.34, p<0.001).

Oral cancer is a major global health concern, ranking sixth worldwide and causing an estimated 330,000 deaths annually. Two-thirds of cases occur in developed countries. In India, oral cancer is responsible for 30-50% of all malignancies, and India ranks first among males and third among females globally. A male predominance is observed (ratio of 2.2:1). Most oral squamous cell carcinomas (OSCC) appear in the fourth and fifth decades of life, often at leukoplakia sites, though developing countries report its appearance at erythroplakia sites.11,12 The most common locations affected are the tongue (posterior lateral and ventral border), buccal mucosa, floor of mouth, labial mucosa, retromolar area, lateral border of the tongue, and palate. Smokeless tobacco use is more prevalent than smoking in both genders. A significant genetic predisposition is recognized, involving point mutations, gene amplifications, rearrangements, and deletions. Some individuals inherit an impaired ability to metabolize carcinogens or repair DNA damage. Tobacco use, particularly betel quid, tobacco with lime, beedi, and hookah, is a major etiological factor. Smoking and chewing tobacco remain the leading causes of OSCC, with DNA damage potentially caused by methylating agents, reactive oxygen species, and betel quid intermediates. Clinically, OSCC typically appears as a proliferative growth or ulcerated lesion with a necrotic core and folded margins; induration is a sign of invasion. Early-stage oral cancers are often asymptomatic, resulting in more than half of diagnoses occurring at an advanced stage, with a five-year survival rate between 10-40%. Lower survival is attributed to chronic disease progression, inadequate knowledge of molecular pathogenesis, and delayed detection. Evidence from clinical, laboratory, and morphologic studies suggests that non-invasive squamous mucosal lesions progress to high-grade dysplasia and carcinoma in situ. The majority of oral cancers are squamous cell carcinoma, and numerous studies focus on diagnosis, prognosis, and treatment strategies. Epithelial-mesenchymal transformation (EMT), which involves the conversion of epithelial cells to highly mobile mesenchymal cells, contributes to invasion, metastasis, and poor prognosis. This study analyzed E-cadherin and p53 markers regarding tumor differentiation and nodal metastases.13 Gender distribution in this study showed male predominance, with males constituting 77% of cases and females 23%, a male-to-female ratio of 3.28:1. These findings align with previous studies reporting similar gender ratios.14-17 Male predominance is attributed to higher rates of tobacco, paan, gutkha, and alcohol use. Studies on p53 in OSCC also report more cases among males. There was no significant correlation between gender and E-cadherin positivity. The majority of patients (35%) belonged to the 51–60 years age group, with a mean age of 49.81 ± 10.56 years. This distribution is consistent with other studies, where most cases occur between 51–70 years or older than 50 years. With increasing age, the number of E-cadherin positive cases rises.17-21 Regarding lesion sites, 40% were on the tongue, 30% on the buccal mucosa, 13% on the hard palate, 5% each on soft palate, uvula, and retromolar trigone, and less common sites included anterior pillar, lips, and floor of the mouth. These findings correspond with previous studies in site distribution.22-24 Correlating E-cadherin positivity with lesion sites, the tongue was most frequent among positive cases, followed by buccal mucosa and hard palate. The p value for association between E-cadherin and lesion site was not significant. The relationship between p53 positivity and lesion sites similarly favored the tongue and buccal mucosa. The most common habit among patients was smokeless tobacco use, followed by smoking, with occasional mixed habits; no significant association was found between habits and E-cadherin expression. Ethnic and geographic differences influence risk factor profiles and marker expression. Lifetime alcohol use was substantially linked to p53 expression, whereas betel quid chewing and tobacco showed a decreasing risk of p53 expression with higher exposure. E-cadherin was positive in 82.85% of cases. Loss of E-cadherin is associated with reduced cell adhesion, increased invasion, and metastasis, particularly in poorly differentiated tumors. Most well-differentiated squamous cell carcinomas showed higher E-cadherin expression compared to less differentiated tumors. p53 was positive in 77% of cases. Increased p53 expression correlates with higher tumor grade, DNA damage, and malignant transformation. Most OSCC cases exhibit mutant p53, which hinders normal cell cycle control and may promote tumor progression and cell proliferation—mutations are detectable in the majority of OSCC cases. The study cohort included 65.71% well differentiated, 25.71% moderately differentiated, and 8.5% poorly differentiated tumors, consistent with previous studies22-24 showing variation in the prevalence of histological grades. Expression of E-cadherin was statistically higher in well differentiated cases, while p53 expression was more frequent in poorly differentiated tumors and associated with nodal metastasis. Co-evaluation of E-cadherin and p53 suggests that wild-type p53 may upregulate E-cadherin and suppress EMT transcription factors; therefore, combined expression analysis might improve prognostic assessment. No significant association was found between gender or tumor site and marker expression, but advanced age, poor differentiation, and nodal metastasis were strongly associated with reduced E-cadherin and increased p53 positivity. These results highlight the utility of E-cadherin and p53 markers in prognosis and management of OSCC.

In well-differentiated tumours, 57% were positive for E-Cadherin and 46% were positive for p53. E-cadherin levels were lower in neoplastic oral epithelium than in normal oral epithelium. p53 levels increased as pathological and histological grade increased. E-cadherin is utilised to identify individuals at high risk of cancer progression, tumour invasiveness, metastasis, prognosis, and recurrence. P53 expression was found to be significantly correlated with tumour grade.

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