Investigating the expression patterns of Androgen Receptor in different grades of Oral Squamous Cell carcinoma 

Investigating the expression patterns of Androgen Receptor in different grades of Oral Squamous Cell carcinoma | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Investigating the expression patterns of Androgen Receptor in different grades of Oral Squamous Cell carcinoma Nousheen Faizullah Khan, Rafat Amin, Neha Baqai, Syeda Noureen Iqbal, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5387823/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Oral squamous cell carcinoma (OSCC) is a common malignant tumor, in the head and neck region with a high incidence and prevalence rate globally. Oral cancer is one of the most deadly and common cancers among men in Asian nations including Bangladesh, India, and Pakistan. Certain viral infections, alcohol, betel quid, areca nut, and tobacco use have all been linked to the development of OSCC. Although the role of hormones and their receptors have been explored in many types of cancers, in OSCC it is still poorly explored. Previous studies have reported that sex hormone androgens and androgen receptors (AR) are involved in growth and progression of breast and prostate cancers. However their role in OSCC remains controversial. The study aims to understand the expression patterns of the androgen receptor at the transcriptional and translational levels in various grades of patients with OSCC. Further to that the expression of AR changes in response to testosterone was also compared in OSCC parients in relation to healthy population. After obtaining consent, the participants were divided into four categories: healthy, poorly differentiated (PD), moderately differentiated (MD), and well differentiated (WD) squamous cell carcinoma (SCC). By creating an appropriate questionnaire, the patient's history was acquired. Samples of blood and tissue were obtained and processed, for testosterone level measurement using ELISA and RNA and protein estimation using commercially available kits, respectively. Tissue samples were visualized using H&E staining. We found that PDSCC group has highest expression levels of AR mRNA, followed by MDSCC and WDSCC. AR expression and serum testosterone levels were found to have moderately significant association in all subjects, determined by Spearman’s correlation. All groups showed full length AR expression while the splice variant AR-V7 was detected exclusively in MDSCC and PDSCC. This is suggestive of AR-V7 role in in OSCC progression. Higher levels of serum testosterone in OSCC patients than in healthy people provide an evidence for the carcinogenic potential of AR in OSCC. OSCC androgen testosterone androgen receptors squamous oral cancer. Figures Figure 1 Figure 2 Figure 3 Figure 4 INTRODUCTION Head and neck cancer (HNC) is the seventh most frequent disease worldwide, with about 660,000 new cases and 325,000 fatalities per year. 90% of HNCs are squamous cell carcinomas, which develop from the pharynx, larynx, and oral cavity's epithelial lining (Gormley et al., 2022 ). These oral squamous cell carcinomas (OSCC) have high prevalence in Asia (45% in India and 10% in Pakistan). According to Qureshi et al. the rising prevalence in Pakistan has made it the second most frequent malignancy in women and the most common malignancy in males. Karachi is one of the Pakistani cities with a noticeably higher OSCC rate (Qureshi et al., 2016 ). From 2017 to 2019, Karachi reported that 16.68% of adult cancer cases were oral (Pervez et al., 2020 ). It is mostly caused by frequent alcohol use, heavy chewing tobacco use, hereditary susceptibility, low socioeconomic level, poor dental health, diets lacking in antioxidants, and viral infections (Nagao et al., 2009). Despite of recent advancement in diagnostics and cancer treatment, oral cancer has five year survival rate (Taghavi et al., 2015). From 2004 till 2009, around three million newly identified cases of oral and oropharyngeal neoplasm were reported with 70000 death (Sharma et al., 2010 ). These alarming numbers has interested researchers to deeply understand this multifactorial disease. Apart from the aforementioned proven risk factors, extant research has also documented the role of sex hormones, namely testosterone, estrogen, and progesterone, together with their corresponding receptors, in the onset and advancement of OSCC (Mannarin et al. 2009). The carcinogenic effect of androgen and androgen receptor has also been studied among these sex hormones in a number of cancer types, such as breast and prostate cancer, but its significance in OSCC has received less attention (Contaldo et al., 2020 ). Testosterone is one of the androgens, or male sex hormones. The testes and adrenal glands release testosterone in males, whereas the ovaries secrete it in females. The androgen receptor is a nuclear transcription factor associated with steroids that binds to its ligand, testosterone, to control the expression of genes downstream. The AR gene consist of eight exons and seven introns, located on the X chromosome (Xp11–12), encoding four structural domains. These are N-terminal domain followed by highly conserved DNA- binding domains consisting of two zinc finger motifs of four cysteine residues in each, short hinge region and a conserved ligand-binding domain at C- terminal end. The androgen receptor is a 919 amino acids ligand-inducible protein with molecular weight of 110 kDa. It is present inside the cytoplasm. When androgens attach to the AR, the protein undergoes a conformational shift, becomes phosphorylated, and moves as a homodimer into the nucleus as part of the androgen/AR complex. After dimerization, AR attaches to target genes' androgen response elements to modify gene transcription in conjunction with other co-regulators (Čonkaš et al., 2023 ). Studies conducted on patients and in vitro experiments have demonstrated that AR, which is necessary for the proliferation of tumor cells, is expressed by OSCC cells (Marocchio et al., 2013 ; Wu et al., 2015 ). According to Wu et al., premalignant and OSCC tissue expressed more AR than benign tumor tissue. He added that there was increased expression of AR mRNA and proteins, both of which are essential for OSCC cell proliferation and progression (Wu et al., 2015 ). In this present study, we measure blood testosterone levels, assess androgen receptor expression levels at the mRNA and protein levels in various grades of patients with OSCC and in healthy persons, and investigate the relationship between serum testosterone levels and AR expression levels in OSCC. MATERIAL AND METHODS Research Design A total 60 participants were included in the study divided in to four groups of 15 each. The study included only male participants which were grouped as healthy, WDCCS, MDSCC, PDSCC. Tissues and blood samples were collected after an informed consent. Following obtaining consent, the details of the participants including demographic, health status, oral hygiene, oral addiction habits and pathological characteristics of OSCC were collected on a structured questionnaire either from directly questioning the participants, their attendant or participant’s medical records form the hospital Sample Collection For control group, Tissue specimens and blood samples were collected from Dow international Dental College at Dow University of Health Sciences, attended the clinic for third molar extraction. For OSCC patients, tissue and blood samples were collected from department of otorhinolaryngogy civil hospital Karachi. All the experimental work for this study was done at Dow Research Institute of Biotechnology and Basic Sciences (DRIBBS). 30µg of fresh oral tissue specimens were collected and stored at -70°C for extraction of RNA and proteins. Tissue samples for imaging were collected in containers containing formalin. 2ml of blood was also collected for testosterone quantification. Biochemical Investigation Blood sample was centrifuged at 2500g at 4 o C for 15 minutes to separate serum and stored at -70°C until analysis. Levels of Testosterone were quantified using DIAsource testosterone ELISA kit (catalogue #KAPD1559) for in vitro diagnostic testing, following manufacturer’s protocol. RNA and Protein Extraction The All-In-One DNA/RNA/Protein Mini-Preps Kit (Cat# BS88003 BioBasic) was used for RNA and protein extraction from a single biological specimen, as per manufacture specified. The resultant RNA was proceeded for cDNA synthesis and qRT-PCR, after checking the purity and quantity using a Nano drop. The protein was precipitated by incubating 10 min with 600ul of supplied PP solution followed by centrifugation at 9000g for 10 minutes. The supernatant was discarded, and the protein pellet was dissolved in 100ul PBS. the dissolved protein was kept − 80°C for western blotting. cDNA synthesis and qPCR RevertAid First Strand cDNA Snthesis Kit by Thermo Scientific (Cat#K1622) was used to synthesized first strand cDNA from DNAse treated RNA in a total reaction volume of 20ul, according to manufacturs instructions. After the synthesis of cDNA, the samples were subjected for quantitative real time PCR on Qunat-7 Flex Real time PCR Detection System by using SYBR green master mix (cat#A25742). Fast chemistry RNAse free 96 well plates with adhesive films were used for amplification using sequence specific primers designed by Primer Express3 (Table 2.1 ) software and at following cycling conditions 40 cycles at 95°C for 1 sec, 60 o C for 1 sec, 72 o C for 1 sec. GAPDH was used as indigenous control and comparative expression levels of Androgen receptor were calculated by 2−∆∆ Ct method. Reactions were set in triplicates and mean Ct values were used to analyze the expression profile. Table 2.1 Sequence of primers for RT-qPCR S.No Primer Sequence 1. GAPDH (F) 5’-AGGGCTGCTTTTAACTCTGGT-3’ 2. GAPDH (R) 5’-CCCCACTTGATTTTGGAGGGA-3’ 3. AR (F) 5’-CCTGGCTTCCGCAACTTACAC-3’ 4. AR (R) 5’-GGACTTGTGCATGCGGTACTC-3’ Western Blotting The extracted proteins were resolved using 10% SDS-PAGE gels under reducing conditions. Separated proteins from gels were transferred to nitrocellulose membranes. 5% milk was added to the blots for blocking. The blots were then incubated overnight at 4°C with anti-human primary antibody (rabbit/anti-mouse) against AR (Cloud-clone corp. PAB252Ra01). Afterwards, they were incubated with secondary antibody for 45 minutes and washed, the membranes were treated with blocking solution for ten minutes. Finally, substrates (peroxidase and luminol) solutions were applied to the membrane. The individual signals and their matching band intensities were assessed, using Chemidoc (Biored) (Roman et al., 2013 ). Statistical analysis SPSS vs 24.0 was used for statistical analysis, examining continuous and categorical variables. To verify normality of data Shapiro-Wilk's test was used. Associations with tumor grade was analyzed using Chi-Square and Fisher's exact tests. One way Anova was performed to compare blood testosterone levels between controls and OSCC groups. We used the post hoc Dunnet test to do multiple comparisons between the groups. P-value less than 0.05 was deemed statistically significant. RESULTS Demographic characteristics of study participants Data on the research participants' demographics with varying OSCC grades have previously been reported (Baqai et al., 2024 ). In summary, the majority of the participants, who were primarily between the ages of 36 and 50, had a mean age of 46.55 years and a BMI of 22.39 ± 4.5. There were people from a variety of ethnic origins, with Urdu speakers making up the majority (n = 32; 53.3%). There was minimal difference (p < 0.05) in age, BMI, and ethnicity between OSCC patients and healthy individuals. Socioeconomically, n = 22(36.7%) people were poor, while 58.3% of people belonged to the middle class (n = 35). Most participants n = 40(66.7%) were educated at different levels. Blood profile of study participants The ABO blood profile information was provided by the study participants, displayed in Table 3.1 . We found that blood types of healthy people and OSCC patients are not significantly correlated. Blood groups B and O are equally prevalent (n = 15; 25%). Blood group A was present in WDSCC n = 5 (38.7%) and healthy individuals. Blood group O was more prevalent in OSCC patients, PDSCC group n = 7 (46.7%). A limitation to draw any conclusion is that 28.3% of participants n = 17 unaware of their blood group. (Table 3.1 ) Table 3.1 Blood Profile of All Study Participants Variable OSCC Patients’ Grades P-value Total n = 60 Healthy n = 15 WDSCC n = 15 MDSCC n = 15 PDSCC n = 15 n (%) n (%) n (%) n% n % Blood Group A 13(21.7) 6(40.0) 5(33.3)) 1(6.7) 1(6.7) 0.135ˆ B 15(25.0) 2(13.3) 4(26.7) 4(26.7) 5(33.3) O 15(25.0) 2(13.3) 2(13.3) 4(26.7) 7(46.7) Don’t know 17(28.3) 5(33.3) 4(26.7) 6(40.0) 2(13.3) *P value was calculated using *chi-square and ^ Fisher’s exact test and p. value < 0.05 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamouse cell carcinoma = PDSCC Clinico-Pathological Features of Study Participants We observed no significant relationship between co-morbidities and study groups, with 33.3% experiencing ear pain, 3.3% diabetic and CVD, and 1.7% OSCC patient reporting renal complications (Table 3.2 ). Table 3.2 Clinico-Pathological features of Study Participants. Variable OSCC Patients ‘Grade P-value Total n = 60 Healthy n = 15 WDSCC n = 15 MDSCC n = 15 PDSCC n = 15 n (%) n (%) n (%) n (%) n (%) Diabetes Mellitus Yes 2(3.3) 0(0.0) 0(0.0) 1(6.7) 1(6.7) 1.00ˆ No 58(96.7) 15(100) 15(100) 14(93.3) 14(93.3) Cardiovascular Disease Yes 2(3.3) 2(13.3) 0(0) 0(0.0) 0(0.0) 0.237ˆ No 58(96.7) 13(86.7) 15(100) 15(100) 15(100) Kidney Disease Yes 1(1.7) 0(0.0) 0(0.0) 1(6.7) 0(0.0) 1.00ˆ No 59(98.3) 15(100) 15(100) 14(93.3) 15(100) Pain in Ear Yes 20(33.3) 1(6.7) 8(53.3) 6(40.0) 5(33.3) 0.066* No 40(66.7) 14(93.3) 7(46.7) 9(60.0) 10(66.7) Any Other Disease Yes 2(3.3) 1(6.7) 0(0.0) 1(6.7) 0(0.0) 1.00ˆ No 58(96.7) 14(93.3) 15(100) 14(93.3) 15(100) *P value was calculated using *chi-square and ^ Fisher’s exact test and p. value < 0.05 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamous cell carcinoma = PDSCC Oral Profile of Study Participants Based on the questions given (supplementary material), Table 3.3 shortlisted the participants' oral health profiles. 56.7% participants brushed their teeth once daily and 33.3% brushed twice daily. We found that oral health practices of OSCC participants and healthy groups were strongly correlated (p = 0.019). 28.3% of participants complained about toothache while 41.2% were in good oral health. According to their medical history, molar extraction had been performed in only 26.7% participants involved in study. Table 3.3 Oral Profile of Study Participants Variable OSCC Patients’ Grades P-value Total n = 60 Healthy n = 15 WDSCC n = 15 MDSCC n = 15 PDSCC n = 15 n (%) n (%) n (%) n (%) n (%) Brushing Habits Once 34(56.7) 9(60.0) 10(66.7) 12(80.0) 3(20.0) 0.019ˆ Twice 20(33.3) 5(33.3) 3(20.0) 2(13.3) 10(66.7) None 6(10.0) 1(6.7) 2(13.3) 1(6.7) 2(13.3) Toothache Yes 17(28.3) 7(46.7) 1(6.7) 5(33.3) 4(26.7) 0.098* No 43(71.7) 8(53.3) 14(93.3) 10(66.7) 11(73.3) Previous Molar Extraction Yes 16(26.7) 4(26.7) 5(33.3) 6(40.0) 1(6.7) 0.192 No 44(73.3) 11(73.3) 10(66.7) 9(60.0) 14(93.3) *P value was calculated using *chi-square and ^ Fisher’s exact test and p. value < 0.05 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamous cell carcinoma = PDSCC Oral Addiction Habits of Study Participants Oral habits of participants are summarized in Table 3.4 . Majority of the participants (71.1%) were consumers of smokeless tobacco (gutka, pan, chalia). 30.0% of individuals were smokers exclusively, while 29% used multiple types, with chalia and gutka being the most common. From the data obtained, we found a significant correlation between OSCC groups and healthy individuals (p = 0.00). Table 3.4 Oral Addiction Habits of Study Participants Variables OSCC patient’s Grades P-value Total n = 60 Healthy n = 15 WDSCC n = 15 MDSCC n = 15 PDSCC n = 15 n (%) n (%) n (%) n (%) n (%) Smoking 0.05ˆ Yes 18(30.0) 1(6.7) 8(53.3) 4(26.7) 5(33.3) No 42(70.0) 14(93.3) 7(46.7) 11(73.3) 10(66.7) Smokeless Tobacco 0.000ˆ Yes 43(71.7) 3(20.0) 12(80.0) 15(100.0) 13(86.7) No 17(28.3) 12(80.0) 3(20.0) 0(0.0) 2(13.3) Type of Smokeless tobacco Gutka 8(19.5) 0(0.0) 2(16.7) 5(38.5) 1(7.7) 0.05ˆ Supari 3(7.3) 0(0.0) 0(0.0) 0(0.0) 3(23.1) Paan 5(12.2) 0(0.0) 3(25.0) 1(7.7) 1(7.7) Naswar 5(12.2) 1(33.3) 1(8.3) 1(7.7) 2(15.4) Chalia 8(19.5) 2(66.7) 0(0.0) 2(15.4) 4(30.8) More than one 12(29.3) 0(0.0) 6(50.0) 3(30.8) 2(15.4) *P value was calculated using *chi-square and ^ Fisher’s exact test and p. value < 0.05 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamous cell carcinoma = PDSCC Clinico-Pathological Characteristics of Oral Cavity in OSCC Patients The clinical characteristics of OSCC participants' oral cavity, was noted. These featured oral lesions, lumps, indifferent patches, chewing or swallowing difficulties, and restricted mouth opening (Table 3.6 ). Results showed oral lesion was the most prevalent characteristic in all grades (93.3%), followed by indifferent patches (80%), chewing or swallowing difficulties, (80%), restricted mouth opening (64.4%) and lumps (17.8%). From our observation significant relationship between oral cavity clinico-pathological features and OSCC grades is lacking (Table 3.5 ). Table 3.5 Clinico-Pathological characteristics of oral cavity in OSCC patients. Variable P-value Total n = 45 WDSCC n = 15 MDSCC n = 15 PDSCC n = 15 n (%) n (%) n (%) n (%) Any Oral Lesion Yes 42(93.3) 14(93.3) 13(86.7) 15(100.0) 0.762ˆ No 3(6.7) 1(6.7) 2(13.3) 0(0.0) Oral Lump Yes 8(17.8) 3(20.0) 3(20.0) 2(13.3) 1.000ˆ No 37(82.2) 12(80.0) 12(80.0) 13(86.7) White or Red Patch in Mouth Yes 36(80.0) 12(80.0) 12(80.0) 12(80.0) 1.000ˆ No 9(20.0) 3(20.0) 3(20.0) 3(20.0) Trouble in chewing or Swallowing Yes 36(80.0) 11(73.3) 11(73.3) 14(93.3) 0.340ˆ No 9(20.0) 4(26.7) 4(26.7) 1(6.7) Restricted mouth opening Yes 29(64.4) 9(60.0) 8(53.3) 12(86.0) 0.387* No 16(35.6) 6(40.0) 7(46.7) 3(20.0) *P value was calculated using *chi-square and ^ Fisher’s exact test and p-value < 0.05 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamous cell carcinoma = PDSCC Hereditary Profile of OSCC Patients Previously we have reported that the tongue (26.7%) and buccal mucosa (cheek) (55.6%) are the two most often implicated tumour sites (Baqai et al., 2024 ). Here the hereditary profile of OSCC patients is displayed in Table 3.7 . 44 individuals had no history available, and just one patient claims having had PDSCC in the past. No discernible relationship between the various tumor grades and family history (p > 0.05) was found. Table 3.6 Family history of OSCC Variable Tumor Grade P-value Total n = 45 WDSCC(n = 15) MDSCC(n = 15) PDSCC(n = 15) n(%) n% n% n=% Available 1(6.7) 0(0.0) 0(0.0) 1(100) 0.89ˆ Unavailable 44(93.3) 15(34.4) 15(34.4) 14(13.8) *P value was calculated using *chi-square and ^ Fisher’s exact test and p-value < 0.05 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamous cell carcinoma = PDSCC Association of Serum Testosterone levels and AR Expression As determined by Spearman's correlation (Table 3.7 ; Fig. 3.1 ) Healthy (r = 0.414; p = 0.126) and WDSCC (r = 0.232; p = 0.425) participants had a weakly non-significant correlation with AR expression, while MDSCC (r = 0.657; p = 0.008) and PDSCC (r = 0.682; p = 0.005) patients showed a moderate positive correlation. Overall, serum testosterone levels and androgen receptor expression in healthy individuals and different grades of OSCC patients showed significantly moderate positive correlation (r = 0.568; p = 0.000). Table 3.7 Association of Serum testosterone levels with AR expression Expression Testosterone levels Overall Healthy WDSCC MDSCC PDSCC r = 0.568 r = 0.414 r = 0.232 r = 0.657 r = 0.682 p = 0.000 p = 0.126 p = 0.425 p = 0.008* p = 0.005* r = Spearman’s correlation and p-value < 0.01 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamous cell carcinoma = PDSCC Blood Serum Testosterone Levels of Study Participants Biochemical assays were performed to quantify the individuals' blood testosterone levels. Statistical analysis (Shapiro-Wilk test) of the obtained data showed that testosterone levels were normal across the research groups (P > 0.05). Variance and means were examined using parametric testing. A significant variation in testosterone level for all four groups can be seen (Table 3.8 , Fig. 3.2 ) (P. Value0.001). According to Dunnett's test, as histological grades rose, mean testosterone levels in WDSCC (mean difference = 14.49, P-value ˂ 0.001), MDSCC (mean difference = 16.90, P-˂0.001), and PDSCC (mean difference = 16.58, P-˂0.001) increased considerably in comparison to the control group. The study highlights how important it is to understand and manage testosterone levels in various cancer types. Table 3.8 Multiple comparison (Dunnet test) between different groups and Testosterone levels Tumor Grade Absolute Mean Difference P-value Healthy vs WDSCC 14.49 ˂0.001 Healthy vs MDSCC 16.90 ˂0.001 Healthy vs PDSCC 16.58 ˂0.001 *P value was calculated using *chi-square and ^ Fisher’s exact test and p-value < 0.05 was considered significant. Well differentiated squamous cell carcinoma = WDSCC, Moderately differentiated squamous cell carcinoma = MDSCC, Poorly differentiated squamous cell carcinoma = PDSCC Histopathological Grading of OSCC Patients Histopathological examination of H&E stained tissue samples was done previously. The samples were classified according to the cancer grading methodology into WDSCC, MDSCC, and PDSCC (Baqai et al., 2024 ). AR Expression in Healthy Oral Tissue and OSCC Tissue through RT-PCR The transcript levels of AR produced in healthy and OSCC was quantified using Real-time qPCR. As shown in Fig. 3.3 expression of AR mRNA was higher in oral squamous cell carcinoma (OSCC) tissues in contrast to healthy tissues. Moreover, significantly higher AR mRNA expression can be seen in PDSCC tissues as compared to MDSCC and WDSCC. Expression of Androgen Receptor Protein in Healthy Subjects and OSCC Patients The Androgen Receptor protein formed in tissue samples from OSCC patients and healthy persons were visualized and compared. AR protein was fully expressed in healthy controls and WDSCC group. A band of full length AR 110kDa can be seen on blots of healthy and WDSCC (Fig. 3.4 ). AR protein expressed in MDSCC and PDSCC groups appeared as band near 75kDa protein marker, suggesting expression of AR protein's V7 splice variant. DISCUSSION Oral squamous cell carcinoma (OSCC) is one of the highly prevalent malignancy in developing counties like India and Pakistan (Kim et al., 2016 ). The disease is asymptomatic at early and advanced stages, making it difficult to detect and treat. It is a multifactorial disease and the most contributing factors include excessive tobacco use, alcohol intake, areca, betel quid nut, ultraviolet radiation, infectious agents, oxidative stress, and compromised DNA repair mechanisms (Sabir et al., 2012 ). Human sex hormones, particularly androgen and androgen receptor, have been studied in various types of cancer, such as breast and prostate cancer (Yeap et al., 1999 ). Therefore their role in OSCC has also been speculated (Mannarini et al., 2009 ). Androgen is a major steroidal sex hormone secreted by the testes, ovaries, and adrenal glands. It regulates various body functions, particularly in males. Androgen receptors have various functional domains and are located on chromosomes Xq11-12. When bound to a ligand, AR modulates the expression of androgen-targeted genes by controlling DNA transcription (Yong et al., 1998 ). The expression levels of androgen receptors vary across various tissues, but higher expression levels are observed in the prostate, liver, and adipocytes (Sar et al., 1990 ). The expression pattern of androgen receptors in OSCC cells has not been analyzed at different histological grades at mRNA and protein levels, and serum testosterone levels have not been evaluated in OSCC. Here in this study, we examined and compared androgen receptor expression patterns in OSCC patients and healthy individuals, and analyzed serum testosterone levels. The study involved 60 male participants divided into four groups: healthy, well differentiated, moderately differentiated, and poorly differentiated squamous cell carcinoma. OSCC effects patients that aged from 36–50 years, with a mean age of 45.6. This is coherent with previous studies that show high prevalence of OSCC in younger populations (Abdulla et al., 2018 ; Alamgir et al., 2016 ; Mahmood et al., 2018 ). However, Mirza et al. reported that mostly patients were diagnosed at advanced age (Mirza et al., 1996). We found that most enrolled participants have a normal BMI and no significant difference in BMI between healthy and OSCC patients was noted. Hence, OSCC didn't affect the BMI of an individual (Baqai et al., 2024 ). Moreover, the majority of participants were Urdu speaking (53.3%), because it is predominant population in Karachi followed by Sindhi (Baqai et al., 2024 ). This is consistent with previous studies in Karachi, where oral cancer prevalence is higher among Urdu-speaking populations (Johnson et al., 2011 ; Mazahir et al., 2006 ; Shumaila et al., 2016 ; Mehdi et al., 2018 ).we also found our research is that education levels between healthy and OSCC patients differ significantly, as expected OSCC patients are mostly less educated compared to healthy participants. From our results we observed that blood group B and O are the most frequent blood groups among OSCC patients, with blood group B being more prevalent in OSCC. Similar results have been documented previously where blood group B was more prevalent in OSCC (Poornima et al., 2018 ; Mortazavi et al., 2014 ). The biochemical cause for it is yet to be deduced. Poor oral hygiene is linked to oral cancer, as it enhances the carcinogenic effects of other risk factors like tobacco. Most participants have once a day tooth brushing habit, which is consistent with previous studies (Mathur et al., 2019 ; Balaram et al., 2002 ; Guha et al., 2007 ). OSCC patients had a higher history of loose teeth, suggesting a connection between loose teeth and poor oral hygiene as well as an increased risk of oral cavity cancer. However the difference was non-significant between both OSCC patients and those in good health. Our findings contradicts previous research that linked smoking with higher OSCC risk. We didn't find any discernable association between smoking habit and risk of oral squamous cell carcinoma (OSCC). It is possibly due to low socioeconomic status in Pakistan compared to western countries (Tanaka et al., 2011). However, in South Asian countries prevalence of chewing tobacco is 22%, while in western countries it is less than 1%. (John et al., 2018 ). This is due to easy availability and low prices of chewing tobacco. Studies by Shenoi et al. and Anwar et al. have shown a significant association between smokeless tobacco and oral cancer, with gutka being the most prevalent type (Shenoi et al., 2012 ; Younas et al., 2016 ; Shumalia et al., 2016). Excessive use of gutka, niswar, and manpuri contributes to the development of OSCC (Tanaka et al., 2011). Low socioeconomic status, poor education, and lack of knowledge about carcinogenic substances may contribute to this prevalence. Oral cancer sites vary widely, with buccal mucosa (41.7%) being the most involved site in oral squamous cell carcinoma (OSCC), followed by the tongue (20.0%). Studies from India and western countries have reported different oral cancer localizations, with the tongue being more frequent in western countries due to excessive alcohol use (Tanaka et al., 2011; Jerjes et al., 2010 ). In our population, excessive chewing tobacco may be the cause of buccal mucosa involvement. Many study reported the presence of oral lesion and leukoplakia as a prominent clinical feature in majority of OSCC patients (Napier et al., 2008). We also found that oral lesions and leukopalakia as the most prevalent clinical symptoms in OSCC patients, with oral lumps found in 17.8% of cases. Here we demonstrated the role of AR in different histological grades of Oropharyngeal Squamous Cell Carcinoma (OSCC) tissues. We found that in different grades of OSCC mRNA is expressed at different concentrations. Higher expression was found in all three OSCC grades (PDSCC, WDSCC and, MDSCC) as compared to healthy individuals. This suggests that altered expression of AR might have a role in the growth and progression of OSCC. Studies have highlighted the importance of AR expression for the growth and progression of OSCC cells, with higher expression levels observed in premalignant and malignant tissue compared to normal mucosa tissue (Wu et al., 2015 ). In prostate and breast cancer, the expression of AR has been well discussed, but its role in OSCC remains unclear. From previous studies it was found that AR expression is detected in 16% of OSCC cases, with significant differences between genders (Marocchio et al., 2013 ; Mohamed et al., 2018 ). In laryngeal carcinoma, AR expression is 68.3%, and in juvenile nasopharyngeal fibroma, it is highly expressed in tumor fibroblast cells (Chen et al., 2006 ; Hagen et al., 1994 ). Fard et al. reported that in gastric cancer, AR is overexpressed in 57 patients at transcriptional level (Fard et al., 2020 ), and in renal carcinoma, it is significantly higher in patients with pathological stage 2 tumors documented by Ha et al. (Ha et al., 2015 ). In contrast to our findings, some studies have reported the opposite role of AR, such as its antitumor role in ER-positive breast cancer stated by Hickey et al. and its low expression in OSCC tissue compared to normal control tissue (Colella et al., 2011 ). The human androgen gene, encoding a full-length 110kda protein, has multiple shortened isoforms lacking functional domains. These are called splice variants (AR-Vs) (Cao et al., 2016 ). These splice variants (ARs-Vs) have the ability to execute transcription, resulting in an uncontrollable tumor growth, even when the androgen signaling is absent (Chan et al., 2015 ; Watson et al., 2010 ). Studies have reported the presence of multiple splice variants of AR with ARVs mutated or truncated at the ligand binding domain.in prostate and breast cancers. The most detected and reviewed AR-Vs is AR-V7 (Watson et al., 2010 ). A direct relation exist between AR expression and serum testosterone levels. Thus, an enhanced testosterone levels can modify AR expression and stimulate cell proliferation in advanced stages of OSCC. We also found a strong correlation of serum testosterone between healthy oral tissues and histologically graded OSCC tissues. In poor and moderately differentiated tissue the testosterone levels were higher as compared to well differentiated OSCC. It implies that as the cancer progressed the testosterone levels rose. This is the first study to simultaneously assess serum testosterone levels in OSCC, as well as expression of AR mRNA and protein. Hence, patients with oral squamous cell carcinoma may have serum testosterone levels that can serve as a potential biomarker for OSCC. Ethical consideration The research project was presented in front of Institutional Review Board (IRB) of DUHS and the ethical approval for the study design, experimentation and ethical matters was granted by the board (REF IRB-1535/DUHS/Approval/2020). CONCLUSION To the best of our knowledge, no literature have been found that investigated the expression of AR mRNA and protein levels and correlation of serum testosterone levels with the expression of AR in OSCC patients. We first time investigated the expression of androgen receptor (AR) mRNA and protein levels and found that it is overexpressed in different histological grades of OSCC as compared to healthy individuals. The study also found a significant correlation between serum testosterone levels and AR expression in OSCC patients, and observed full length androgen receptor and its splice variant AR-V7 in poorly and moderately differentiated samples. Declarations Author Contribution Nousheen Faizullah Khan: sample collection and processing, all protocols included in methodology, compilation of results and prepared all the figures and statistical evaluation, initial drafting of manuscriptRafat Amin: Idea conceptualization, protocol optimization, supervision of the study, manuscript editing and proof readingNeha Baqai: sample collection and processing, protocol optimization, statistical evaluation, manuscript proof readingSyeda Noureen Iqbal: provision of samples, clinical validation of study subjects, manuscript revisionZeba Ahmed: provision of samples, clinical validation of study subjects, manuscript revisionTehseen Fatima: Idea conceptualization, protocol optimization, supervision of the study, manuscript writeup, editing and final proof reading References Abdulla, R., Adyanthaya, S., Kini, P., Mohanty, V., D'Souza, N., & Subbannayya, Y. (2018). 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Demographic and clinical profile of oral squamous cell carcinoma patients: A retrospective study. Indian journal of cancer, 49(1), 21-26. Shumaila, Y., Naila Irum, H., Farah, A., & Haya, M. (2016). Role of chewing habits and cigarette smoking in differentiation of oral squamous cell carcinoma. Taghavi, N., & Yazdi, I. (2015). Prognostic factors of survival rate in oral squamous cell carcinoma: clinical, histologic, genetic and molecular concepts. Arch Iran Med, 18(5), 314-319. Tanaka, T., & Ishigamori, R. (2011). Understanding carcinogenesis for fighting oral cancer. Journal of oncology, 2011. Watson, P. A., Chen, Y. F., Balbas, M. D., Wongvipat, J., Socci, N. D., Viale, A., Kim, K., & Sawyers, C. L. (2010). Constitutively active androgen receptor splice variants expressed in castration-resistant prostate cancer require full-length androgen receptor. Proceedings of the national academy of sciences, 107(39), 16759-16765. Wu, T. F., Luo, F. J., Chang, Y. L., Huang, C. M., Chiu, W. J., Weng, C. F., Hsu, Y. K., & Yuan, T. C. (2015). The oncogenic role of androgen receptors in promoting the growth of oral squamous cell carcinoma cells. Oral Dis, 21(3), 320-327. https://doi.org/10.1111/odi.12272 Yeap, B. B., Krueger, R. G., & Leedman, P. J. (1999). Differential posttranscriptional regulation of androgen receptor gene expression by androgen in prostate and breast cancer cells. Endocrinology, 140(7), 3282-3291. https://doi.org/10.1210/endo.140.7.6769 Yong, E. L., Ghadessy, F., Wang, Q., Mifsud, A., & Ng, S. C. (1998). Androgen receptor transactivation domain and control of spermatogenesis. Rev Reprod, 3(3), 141-144. https://doi.org/10.1530/ror.0.0030141 Younas, S., Hadi, N. I., Ahmed, F., & Mohammad, H. (2016). Role of chewing habits and cigarette smoking in differentiation of oral squamous cell carcinoma. Pak. J. Med, 5(3), 32-37. Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5387823","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":379609928,"identity":"5fc253c0-daf2-44fa-82cc-207542900387","order_by":0,"name":"Nousheen Faizullah Khan","email":"","orcid":"","institution":"School of Post Graduate Studies, Dow university of Health Sciences, Ojha Campus Karachi, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Nousheen","middleName":"Faizullah","lastName":"Khan","suffix":""},{"id":379609929,"identity":"7d3abf66-303b-4fa1-8a4e-e1c4af2dbe4d","order_by":1,"name":"Rafat Amin","email":"","orcid":"","institution":"Dow University of Health Sciences, Dow College of Biotechnology","correspondingAuthor":false,"prefix":"","firstName":"Rafat","middleName":"","lastName":"Amin","suffix":""},{"id":379609930,"identity":"1e6e5564-463d-471b-ad51-45bf0ff4e32d","order_by":2,"name":"Neha Baqai","email":"","orcid":"","institution":"School of Post Graduate Studies, Dow university of Health Sciences, Ojha Campus Karachi, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Neha","middleName":"","lastName":"Baqai","suffix":""},{"id":379609931,"identity":"ac524505-30f8-4e26-b459-3aeb23fd813f","order_by":3,"name":"Syeda Noureen Iqbal","email":"","orcid":"","institution":"Oral and Maxillofacial Surgery Department, DIEKIOHS, Ojha Campus, Dow university of Health Sciences.","correspondingAuthor":false,"prefix":"","firstName":"Syeda","middleName":"Noureen","lastName":"Iqbal","suffix":""},{"id":379609932,"identity":"ac5cbccf-92fa-4223-b3fe-07eb7c2bb5c0","order_by":4,"name":"Zeba Ahmed","email":"","orcid":"","institution":"Dow Medical College-Dr.Ruth KM Pfau Civil Hospital Karachi, Dow University of Health Sciences","correspondingAuthor":false,"prefix":"","firstName":"Zeba","middleName":"","lastName":"Ahmed","suffix":""},{"id":379609933,"identity":"96086f7a-b05b-4edf-97ed-cad9988a65f2","order_by":5,"name":"Tehseen Fatima","email":"data:image/png;base64,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","orcid":"","institution":"Dow University of Health Sciences, Dow College of Biotechnology","correspondingAuthor":true,"prefix":"","firstName":"Tehseen","middleName":"","lastName":"Fatima","suffix":""}],"badges":[],"createdAt":"2024-11-04 11:53:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5387823/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5387823/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":70510892,"identity":"eade8497-0620-433a-ba20-b42b7ee6a968","added_by":"auto","created_at":"2024-12-04 00:38:34","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":20432,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 3.1 : Correlation of serum testosterone levels and AR expression\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5387823/v1/d9ad5eb2cb4c25db4fc4ec51.png"},{"id":70510434,"identity":"8d10db6b-c39b-434e-a163-ff3ea5c174f6","added_by":"auto","created_at":"2024-12-04 00:30:34","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":16726,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 3.2: Serum testosterone levels among the study participants.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher’s exact test and p-value \u0026lt;0.05 was considered significant. Well differentiated squamous cell carcinoma= WDSCC, Moderately differentiated squamous cell carcinoma= MDSCC, Poorly differentiated squamous cell carcinoma= PDSCC\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5387823/v1/85e5f506d231d7ab133afbf7.png"},{"id":70510432,"identity":"01ac9b8a-6913-45d4-a23e-828a11577b98","added_by":"auto","created_at":"2024-12-04 00:30:34","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":21799,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 3.3: Expression profiling of Androgen receptor in different histological grades of OSCC\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5387823/v1/f53d778ceaf3dbbee99f386c.png"},{"id":70510435,"identity":"73d14e23-2d2c-4708-9834-46b1b9182207","added_by":"auto","created_at":"2024-12-04 00:30:34","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":331610,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFigure 3.4: \u003c/strong\u003eWestern blot analysis of three sets of representative samples of Healthy controls and different histopathological grades of OSCC patients.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5387823/v1/454ed0171625fa4a05b6eaa4.png"},{"id":71998583,"identity":"7d486f7c-a08d-4112-8085-f33cac8a29be","added_by":"auto","created_at":"2024-12-20 12:54:27","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2160116,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5387823/v1/174bacc2-266b-489a-9f1a-14454f84139f.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Investigating the expression patterns of Androgen Receptor in different grades of Oral Squamous Cell carcinoma ","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eHead and neck cancer (HNC) is the seventh most frequent disease worldwide, with about 660,000 new cases and 325,000 fatalities per year. 90% of HNCs are squamous cell carcinomas, which develop from the pharynx, larynx, and oral cavity's epithelial lining (Gormley et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These oral squamous cell carcinomas (OSCC) have high prevalence in Asia (45% in India and 10% in Pakistan). According to Qureshi et al. the rising prevalence in Pakistan has made it the second most frequent malignancy in women and the most common malignancy in males. Karachi is one of the Pakistani cities with a noticeably higher OSCC rate (Qureshi et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). From 2017 to 2019, Karachi reported that 16.68% of adult cancer cases were oral (Pervez et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). It is mostly caused by frequent alcohol use, heavy chewing tobacco use, hereditary susceptibility, low socioeconomic level, poor dental health, diets lacking in antioxidants, and viral infections (Nagao et al., 2009). Despite of recent advancement in diagnostics and cancer treatment, oral cancer has five year survival rate (Taghavi et al., 2015). From 2004 till 2009, around three million newly identified cases of oral and oropharyngeal neoplasm were reported with 70000 death (Sharma et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThese alarming numbers has interested researchers to deeply understand this multifactorial disease. Apart from the aforementioned proven risk factors, extant research has also documented the role of sex hormones, namely testosterone, estrogen, and progesterone, together with their corresponding receptors, in the onset and advancement of OSCC (Mannarin et al. 2009). The carcinogenic effect of androgen and androgen receptor has also been studied among these sex hormones in a number of cancer types, such as breast and prostate cancer, but its significance in OSCC has received less attention (Contaldo et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTestosterone is one of the androgens, or male sex hormones. The testes and adrenal glands release testosterone in males, whereas the ovaries secrete it in females. The androgen receptor is a nuclear transcription factor associated with steroids that binds to its ligand, testosterone, to control the expression of genes downstream. The AR gene consist of eight exons and seven introns, located on the X chromosome (Xp11\u0026ndash;12), encoding four structural domains. These are N-terminal domain followed by highly conserved DNA- binding domains consisting of two zinc finger motifs of four cysteine residues in each, short hinge region and a conserved ligand-binding domain at C- terminal end. The androgen receptor is a 919 amino acids ligand-inducible protein with molecular weight of 110 kDa. It is present inside the cytoplasm. When androgens attach to the AR, the protein undergoes a conformational shift, becomes phosphorylated, and moves as a homodimer into the nucleus as part of the androgen/AR complex. After dimerization, AR attaches to target genes' androgen response elements to modify gene transcription in conjunction with other co-regulators (Čonkaš et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Studies conducted on patients and in vitro experiments have demonstrated that AR, which is necessary for the proliferation of tumor cells, is expressed by OSCC cells (Marocchio et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Wu et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). According to Wu et al., premalignant and OSCC tissue expressed more AR than benign tumor tissue. He added that there was increased expression of AR mRNA and proteins, both of which are essential for OSCC cell proliferation and progression (Wu et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In this present study, we measure blood testosterone levels, assess androgen receptor expression levels at the mRNA and protein levels in various grades of patients with OSCC and in healthy persons, and investigate the relationship between serum testosterone levels and AR expression levels in OSCC.\u003c/p\u003e"},{"header":"MATERIAL AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eResearch Design\u003c/h2\u003e \u003cp\u003eA total 60 participants were included in the study divided in to four groups of 15 each. The study included only male participants which were grouped as healthy, WDCCS, MDSCC, PDSCC. Tissues and blood samples were collected after an informed consent. Following obtaining consent, the details of the participants including demographic, health status, oral hygiene, oral addiction habits and pathological characteristics of OSCC were collected on a structured questionnaire either from directly questioning the participants, their attendant or participant\u0026rsquo;s medical records form the hospital\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSample Collection\u003c/h3\u003e\n\u003cp\u003eFor control group, Tissue specimens and blood samples were collected from Dow international Dental College at Dow University of Health Sciences, attended the clinic for third molar extraction. For OSCC patients, tissue and blood samples were collected from department of otorhinolaryngogy civil hospital Karachi. All the experimental work for this study was done at Dow Research Institute of Biotechnology and Basic Sciences (DRIBBS). 30\u0026micro;g of fresh oral tissue specimens were collected and stored at -70\u0026deg;C for extraction of RNA and proteins. Tissue samples for imaging were collected in containers containing formalin. 2ml of blood was also collected for testosterone quantification.\u003c/p\u003e\n\u003ch3\u003eBiochemical Investigation\u003c/h3\u003e\n\u003cp\u003eBlood sample was centrifuged at 2500g at 4\u003csup\u003eo\u003c/sup\u003eC for 15 minutes to separate serum and stored at -70\u0026deg;C until analysis. Levels of Testosterone were quantified using DIAsource testosterone ELISA kit (catalogue #KAPD1559) for in vitro diagnostic testing, following manufacturer\u0026rsquo;s protocol.\u003c/p\u003e\n\u003ch3\u003eRNA and Protein Extraction\u003c/h3\u003e\n\u003cp\u003eThe All-In-One DNA/RNA/Protein Mini-Preps Kit (Cat# BS88003 BioBasic) was used for RNA and protein extraction from a single biological specimen, as per manufacture specified. The resultant RNA was proceeded for cDNA synthesis and qRT-PCR, after checking the purity and quantity using a Nano drop. The protein was precipitated by incubating 10 min with 600ul of supplied PP solution followed by centrifugation at 9000g for 10 minutes. The supernatant was discarded, and the protein pellet was dissolved in 100ul PBS. the dissolved protein was kept \u0026minus;\u0026thinsp;80\u0026deg;C for western blotting.\u003c/p\u003e\n\u003ch3\u003ecDNA synthesis and qPCR\u003c/h3\u003e\n\u003cp\u003eRevertAid First Strand cDNA Snthesis Kit by Thermo Scientific (Cat#K1622) was used to synthesized first strand cDNA from DNAse treated RNA in a total reaction volume of 20ul, according to manufacturs instructions. After the synthesis of cDNA, the samples were subjected for quantitative real time PCR on Qunat-7 Flex Real time PCR Detection System by using SYBR green master mix (cat#A25742). Fast chemistry RNAse free 96 well plates with adhesive films were used for amplification using sequence specific primers designed by Primer Express3 (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e2.1\u003c/span\u003e) software and at following cycling conditions 40 cycles at 95\u0026deg;C for 1 sec, 60\u003csup\u003eo\u003c/sup\u003eC for 1 sec, 72\u003csup\u003eo\u003c/sup\u003eC for 1 sec. GAPDH was used as indigenous control and comparative expression levels of Androgen receptor were calculated by \u003csup\u003e2\u0026minus;∆∆\u003c/sup\u003eCt method. Reactions were set in triplicates and mean Ct values were used to analyze the expression profile.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2.1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cb\u003eSequence of primers for RT-qPCR\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS.No\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePrimer\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSequence\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGAPDH (F)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u0026rsquo;-AGGGCTGCTTTTAACTCTGGT-3\u0026rsquo;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e2.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGAPDH (R)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u0026rsquo;-CCCCACTTGATTTTGGAGGGA-3\u0026rsquo;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e3.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAR (F)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u0026rsquo;-CCTGGCTTCCGCAACTTACAC-3\u0026rsquo;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e4.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAR (R)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u0026rsquo;-GGACTTGTGCATGCGGTACTC-3\u0026rsquo;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eWestern Blotting\u003c/h2\u003e \u003cp\u003eThe extracted proteins were resolved using 10% SDS-PAGE gels under reducing conditions. Separated proteins from gels were transferred to nitrocellulose membranes. 5% milk was added to the blots for blocking. The blots were then incubated overnight at 4\u0026deg;C with anti-human primary antibody (rabbit/anti-mouse) against AR (Cloud-clone corp. PAB252Ra01). Afterwards, they were incubated with secondary antibody for 45 minutes and washed, the membranes were treated with blocking solution for ten minutes. Finally, substrates (peroxidase and luminol) solutions were applied to the membrane. The individual signals and their matching band intensities were assessed, using Chemidoc (Biored) (Roman et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eSPSS vs 24.0 was used for statistical analysis, examining continuous and categorical variables. To verify normality of data Shapiro-Wilk's test was used. Associations with tumor grade was analyzed using Chi-Square and Fisher's exact tests. One way Anova was performed to compare blood testosterone levels between controls and OSCC groups. We used the post hoc Dunnet test to do multiple comparisons between the groups. P-value less than 0.05 was deemed statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003eDemographic characteristics of study participants\u003c/h2\u003e\n \u003cp\u003eData on the research participants\u0026apos; demographics with varying OSCC grades have previously been reported (Baqai et al., \u003cspan class=\"CitationRef\"\u003e2024\u003c/span\u003e). In summary, the majority of the participants, who were primarily between the ages of 36 and 50, had a mean age of 46.55 years and a BMI of 22.39\u0026thinsp;\u0026plusmn;\u0026thinsp;4.5. There were people from a variety of ethnic origins, with Urdu speakers making up the majority (n\u0026thinsp;=\u0026thinsp;32; 53.3%). There was minimal difference (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in age, BMI, and ethnicity between OSCC patients and healthy individuals. Socioeconomically, n\u0026thinsp;=\u0026thinsp;22(36.7%) people were poor, while 58.3% of people belonged to the middle class (n\u0026thinsp;=\u0026thinsp;35). Most participants n\u0026thinsp;=\u0026thinsp;40(66.7%) were educated at different levels.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eBlood profile of study participants\u003c/h2\u003e\n \u003cp\u003eThe ABO blood profile information was provided by the study participants, displayed in Table \u003cspan class=\"InternalRef\"\u003e3.1\u003c/span\u003e. We found that blood types of healthy people and OSCC patients are not significantly correlated. Blood groups B and O are equally prevalent (n\u0026thinsp;=\u0026thinsp;15; 25%). Blood group A was present in WDSCC n\u0026thinsp;=\u0026thinsp;5 (38.7%) and healthy individuals. Blood group O was more prevalent in OSCC patients, PDSCC group n\u0026thinsp;=\u0026thinsp;7 (46.7%). A limitation to draw any conclusion is that 28.3% of participants n\u0026thinsp;=\u0026thinsp;17 unaware of their blood group. (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3.1\u003c/span\u003e)\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eBlood Profile of All Study Participants\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eVariable\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eOSCC Patients\u0026rsquo; Grades\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;60\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHealthy\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en%\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en %\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eBlood Group\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13(21.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6(40.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(33.3))\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"4\"\u003e\n \u003cp\u003e0.135\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15(25.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15(25.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7(46.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDon\u0026rsquo;t know\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17(28.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6(40.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher\u0026rsquo;s exact test and p. value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant. Well differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamouse cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003eClinico-Pathological Features of Study Participants\u003c/h2\u003e\n \u003cp\u003eWe observed no significant relationship between co-morbidities and study groups, with 33.3% experiencing ear pain, 3.3% diabetic and CVD, and 1.7% OSCC patient reporting renal complications (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3.2\u003c/span\u003e).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003e\u003cstrong\u003eClinico-Pathological features of Study Participants.\u003c/strong\u003e\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eVariable\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eOSCC Patients \u0026lsquo;Grade\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;60\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHealthy\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eDiabetes Mellitus\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(3.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1.00\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e58(96.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eCardiovascular Disease\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(3.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e0.237\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e58(96.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13(86.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eKidney Disease\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(1.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1.00\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e59(98.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003ePain in Ear\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8(53.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6(40.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e0.066*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e40(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7(46.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9(60.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAny Other Disease\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(3.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1.00\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e58(96.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher\u0026rsquo;s exact test and p. value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant. Well differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eOral Profile of Study Participants\u003c/h2\u003e\n \u003cp\u003eBased on the questions given (supplementary material), Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3.3\u003c/span\u003e shortlisted the participants\u0026apos; oral health profiles. 56.7% participants brushed their teeth once daily and 33.3% brushed twice daily. We found that oral health practices of OSCC participants and healthy groups were strongly correlated (p\u0026thinsp;=\u0026thinsp;0.019). 28.3% of participants complained about toothache while 41.2% were in good oral health. According to their medical history, molar extraction had been performed in only 26.7% participants involved in study.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eOral Profile of Study Participants\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eVariable\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eOSCC Patients\u0026rsquo; Grades\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;60\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHealthy\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eBrushing Habits\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOnce\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e34(56.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9(60.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.019\u0026circ;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTwice\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6(10.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eToothache\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17(28.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7(46.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e0.098*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e43(71.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8(53.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11(73.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003ePrevious Molar Extraction\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6(40.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e0.192\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e44(73.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11(73.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9(60.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher\u0026rsquo;s exact test and p. value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant. Well differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003eOral Addiction Habits of Study Participants\u003c/h2\u003e\n \u003cp\u003eOral habits of participants are summarized in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3.4\u003c/span\u003e. Majority of the participants (71.1%) were consumers of smokeless tobacco (gutka, pan, chalia). 30.0% of individuals were smokers exclusively, while 29% used multiple types, with chalia and gutka being the most common. From the data obtained, we found a significant correlation between OSCC groups and healthy individuals (p\u0026thinsp;=\u0026thinsp;0.00).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab5\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eOral Addiction Habits of Study Participants\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"7\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eVariables\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" colspan=\"3\"\u003e\n \u003cp\u003eOSCC patient\u0026rsquo;s Grades\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;60\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHealthy\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSmoking\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"3\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.05\u0026circ;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18(30.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8(53.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42(70.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7(46.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11(73.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eSmokeless Tobacco\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.000\u0026circ;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e43(71.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15(100.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13(86.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17(28.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eType of Smokeless tobacco\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGutka\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8(19.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(16.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(38.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(7.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"6\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.05\u0026circ;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eSupari\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(7.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(23.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePaan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(12.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(25.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(7.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(7.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNaswar\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5(12.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(33.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(8.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(7.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(15.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eChalia\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8(19.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(66.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(15.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(30.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMore than one\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(29.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6(50.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(30.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(15.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"7\"\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher\u0026rsquo;s exact test and p. value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant. Well differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n \u003ch2\u003eClinico-Pathological Characteristics of Oral Cavity in OSCC Patients\u003c/h2\u003e\n \u003cp\u003eThe clinical characteristics of OSCC participants\u0026apos; oral cavity, was noted. These featured oral lesions, lumps, indifferent patches, chewing or swallowing difficulties, and restricted mouth opening (Table\u003cspan class=\"InternalRef\"\u003e3.6\u003c/span\u003e). Results showed oral lesion was the most prevalent characteristic in all grades (93.3%), followed by indifferent patches (80%), chewing or swallowing difficulties, (80%), restricted mouth opening (64.4%) and lumps (17.8%). From our observation significant relationship between oral cavity clinico-pathological features and OSCC grades is lacking (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3.5\u003c/span\u003e).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab6\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eClinico-Pathological characteristics of oral cavity in OSCC patients.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"6\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eVariable\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" colspan=\"4\"\u003e\u0026nbsp;\u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal n\u0026thinsp;=\u0026thinsp;45\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eWDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eMDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePDSCC\u003c/p\u003e\n \u003cp\u003en\u0026thinsp;=\u0026thinsp;15\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eAny Oral Lesion\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e42(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13(86.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15(100.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e0.762\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eOral Lump\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8(17.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2(13.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1.000\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e37(82.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13(86.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eWhite or Red Patch in Mouth\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e36(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e1.000\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eTrouble\u0026nbsp;in\u0026nbsp;chewing\u0026nbsp;or\u0026nbsp;Swallowing\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e36(80.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11(73.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11(73.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e0.340\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4(26.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eRestricted\u0026nbsp;mouth\u0026nbsp;opening\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e29(64.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9(60.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8(53.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12(86.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e0.387*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16(35.6)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6(40.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7(46.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3(20.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher\u0026rsquo;s exact test and p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant. Well differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n \u003ch2\u003eHereditary Profile of OSCC Patients\u003c/h2\u003e\n \u003cp\u003ePreviously we have reported that the tongue (26.7%) and buccal mucosa (cheek) (55.6%) are the two most often implicated tumour sites (Baqai et al., \u003cspan class=\"CitationRef\"\u003e2024\u003c/span\u003e). Here the hereditary profile of OSCC patients is displayed in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3.7\u003c/span\u003e. 44 individuals had no history available, and just one patient claims having had PDSCC in the past. No discernible relationship between the various tumor grades and family history (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) was found.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab7\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.6\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eFamily history of OSCC\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"6\"\u003e\u003c/colgroup\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eVariable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\"\u003e\n \u003cp\u003eTumor Grade\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"3\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal n\u0026thinsp;=\u0026thinsp;45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWDSCC(n\u0026thinsp;=\u0026thinsp;15)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMDSCC(n\u0026thinsp;=\u0026thinsp;15)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePDSCC(n\u0026thinsp;=\u0026thinsp;15)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003en(%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003en%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003en%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003en=%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAvailable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(6.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0(0.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1(100)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e0.89\u0026circ;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eUnavailable\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e44(93.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(34.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15(34.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e14(13.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher\u0026rsquo;s exact test and p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant.\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eWell differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n \u003ch2\u003eAssociation of Serum Testosterone levels and AR Expression\u003c/h2\u003e\n \u003cp\u003eAs determined by Spearman\u0026apos;s correlation (Table \u003cspan class=\"InternalRef\"\u003e3.7\u003c/span\u003e; Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3.1\u003c/span\u003e) Healthy (r\u0026thinsp;=\u0026thinsp;0.414; p\u0026thinsp;=\u0026thinsp;0.126) and WDSCC (r\u0026thinsp;=\u0026thinsp;0.232; p\u0026thinsp;=\u0026thinsp;0.425) participants had a weakly non-significant correlation with AR expression, while MDSCC (r\u0026thinsp;=\u0026thinsp;0.657; p\u0026thinsp;=\u0026thinsp;0.008) and PDSCC (r\u0026thinsp;=\u0026thinsp;0.682; p\u0026thinsp;=\u0026thinsp;0.005) patients showed a moderate positive correlation. Overall, serum testosterone levels and androgen receptor expression in healthy individuals and different grades of OSCC patients showed significantly moderate positive correlation (r\u0026thinsp;=\u0026thinsp;0.568; p\u0026thinsp;=\u0026thinsp;0.000).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab8\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.7\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAssociation of Serum testosterone levels with AR expression\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"6\"\u003e\u003c/colgroup\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"4\"\u003e\n \u003cp\u003eExpression\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"5\"\u003e\n \u003cp\u003eTestosterone levels\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOverall\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHealthy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWDSCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMDSCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePDSCC\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003er\u0026thinsp;=\u0026thinsp;0.568\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003er\u0026thinsp;=\u0026thinsp;0.414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003er\u0026thinsp;=\u0026thinsp;0.232\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003er\u0026thinsp;=\u0026thinsp;0.657\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003er\u0026thinsp;=\u0026thinsp;0.682\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ep\u0026thinsp;=\u0026thinsp;0.000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ep\u0026thinsp;=\u0026thinsp;0.126\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ep\u0026thinsp;=\u0026thinsp;0.425\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ep\u0026thinsp;=\u0026thinsp;0.008*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ep\u0026thinsp;=\u0026thinsp;0.005*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003e\u003cstrong\u003er\u0026thinsp;=\u0026thinsp;Spearman\u0026rsquo;s correlation and p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.01 was considered significant. Well differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n \u003ch2\u003eBlood Serum Testosterone Levels of Study Participants\u003c/h2\u003e\n \u003cp\u003eBiochemical assays were performed to quantify the individuals\u0026apos; blood testosterone levels. Statistical analysis (Shapiro-Wilk test) of the obtained data showed that testosterone levels were normal across the research groups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Variance and means were examined using parametric testing. A significant variation in testosterone level for all four groups can be seen (Table \u003cspan class=\"InternalRef\"\u003e3.8\u003c/span\u003e, Fig. \u003cspan class=\"InternalRef\"\u003e3.2\u003c/span\u003e) (P. Value0.001). According to Dunnett\u0026apos;s test, as histological grades rose, mean testosterone levels in WDSCC (mean difference\u0026thinsp;=\u0026thinsp;14.49, P-value ˂ 0.001), MDSCC (mean difference\u0026thinsp;=\u0026thinsp;16.90, P-˂0.001), and PDSCC (mean difference\u0026thinsp;=\u0026thinsp;16.58, P-˂0.001) increased considerably in comparison to the control group. The study highlights how important it is to understand and manage testosterone levels in various cancer types.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u0026nbsp;\u003ctable id=\"Tab9\" border=\"1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3.8\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eMultiple comparison (Dunnet test) between different groups and Testosterone levels\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003ccolgroup cols=\"3\"\u003e\u003c/colgroup\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTumor Grade\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eAbsolute Mean Difference\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eHealthy vs WDSCC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e˂0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eHealthy vs MDSCC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e˂0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003eHealthy vs PDSCC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u003cstrong\u003e˂0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\"\u003e\u003cstrong\u003e*P value was calculated using *chi-square and ^ Fisher\u0026rsquo;s exact test and p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered significant. Well differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;WDSCC, Moderately differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;MDSCC, Poorly differentiated squamous cell carcinoma\u0026thinsp;=\u0026thinsp;PDSCC\u003c/strong\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e\n \u003ch2\u003eHistopathological Grading of OSCC Patients\u003c/h2\u003e\n \u003cp\u003eHistopathological examination of H\u0026amp;E stained tissue samples was done previously. The samples were classified according to the cancer grading methodology into WDSCC, MDSCC, and PDSCC (Baqai et al., \u003cspan class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n \u003ch2\u003eAR Expression in Healthy Oral Tissue and OSCC Tissue through RT-PCR\u003c/h2\u003e\n \u003cp\u003eThe transcript levels of AR produced in healthy and OSCC was quantified using Real-time qPCR. As shown in Fig. \u003cspan class=\"InternalRef\"\u003e3.3\u003c/span\u003e expression of AR mRNA was higher in oral squamous cell carcinoma (OSCC) tissues in contrast to healthy tissues. Moreover, significantly higher AR mRNA expression can be seen in PDSCC tissues as compared to MDSCC and WDSCC.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\n \u003ch2\u003eExpression of Androgen Receptor Protein in Healthy Subjects and OSCC Patients\u003c/h2\u003e\n \u003cp\u003eThe Androgen Receptor protein formed in tissue samples from OSCC patients and healthy persons were visualized and compared. AR protein was fully expressed in healthy controls and WDSCC group. A band of full length AR 110kDa can be seen on blots of healthy and WDSCC (Fig. \u003cspan class=\"InternalRef\"\u003e3.4\u003c/span\u003e). AR protein expressed in MDSCC and PDSCC groups appeared as band near 75kDa protein marker, suggesting expression of AR protein\u0026apos;s V7 splice variant.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eOral squamous cell carcinoma (OSCC) is one of the highly prevalent malignancy in developing counties like India and Pakistan (Kim et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The disease is asymptomatic at early and advanced stages, making it difficult to detect and treat. It is a multifactorial disease and the most contributing factors include excessive tobacco use, alcohol intake, areca, betel quid nut, ultraviolet radiation, infectious agents, oxidative stress, and compromised DNA repair mechanisms (Sabir et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eHuman sex hormones, particularly androgen and androgen receptor, have been studied in various types of cancer, such as breast and prostate cancer (Yeap et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1999\u003c/span\u003e). Therefore their role in OSCC has also been speculated (Mannarini et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Androgen is a major steroidal sex hormone secreted by the testes, ovaries, and adrenal glands. It regulates various body functions, particularly in males. Androgen receptors have various functional domains and are located on chromosomes Xq11-12. When bound to a ligand, AR modulates the expression of androgen-targeted genes by controlling DNA transcription (Yong et al., \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). The expression levels of androgen receptors vary across various tissues, but higher expression levels are observed in the prostate, liver, and adipocytes (Sar et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1990\u003c/span\u003e). The expression pattern of androgen receptors in OSCC cells has not been analyzed at different histological grades at mRNA and protein levels, and serum testosterone levels have not been evaluated in OSCC. Here in this study, we examined and compared androgen receptor expression patterns in OSCC patients and healthy individuals, and analyzed serum testosterone levels.\u003c/p\u003e \u003cp\u003e The study involved 60 male participants divided into four groups: healthy, well differentiated, moderately differentiated, and poorly differentiated squamous cell carcinoma. OSCC effects patients that aged from 36\u0026ndash;50 years, with a mean age of 45.6. This is coherent with previous studies that show high prevalence of OSCC in younger populations (Abdulla et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Alamgir et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Mahmood et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). However, Mirza et al. reported that mostly patients were diagnosed at advanced age (Mirza et al., 1996). We found that most enrolled participants have a normal BMI and no significant difference in BMI between healthy and OSCC patients was noted. Hence, OSCC didn't affect the BMI of an individual (Baqai et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Moreover, the majority of participants were Urdu speaking (53.3%), because it is predominant population in Karachi followed by Sindhi (Baqai et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). This is consistent with previous studies in Karachi, where oral cancer prevalence is higher among Urdu-speaking populations (Johnson et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Mazahir et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Shumaila et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Mehdi et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).we also found our research is that education levels between healthy and OSCC patients differ significantly, as expected OSCC patients are mostly less educated compared to healthy participants.\u003c/p\u003e \u003cp\u003eFrom our results we observed that blood group B and O are the most frequent blood groups among OSCC patients, with blood group B being more prevalent in OSCC. Similar results have been documented previously where blood group B was more prevalent in OSCC (Poornima et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Mortazavi et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The biochemical cause for it is yet to be deduced. Poor oral hygiene is linked to oral cancer, as it enhances the carcinogenic effects of other risk factors like tobacco. Most participants have once a day tooth brushing habit, which is consistent with previous studies (Mathur et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Balaram et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Guha et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). OSCC patients had a higher history of loose teeth, suggesting a connection between loose teeth and poor oral hygiene as well as an increased risk of oral cavity cancer. However the difference was non-significant between both OSCC patients and those in good health.\u003c/p\u003e \u003cp\u003eOur findings contradicts previous research that linked smoking with higher OSCC risk. We didn't find any discernable association between smoking habit and risk of oral squamous cell carcinoma (OSCC). It is possibly due to low socioeconomic status in Pakistan compared to western countries (Tanaka et al., 2011). However, in South Asian countries prevalence of chewing tobacco is 22%, while in western countries it is less than 1%. (John et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This is due to easy availability and low prices of chewing tobacco. Studies by Shenoi et al. and Anwar et al. have shown a significant association between smokeless tobacco and oral cancer, with gutka being the most prevalent type (Shenoi et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Younas et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Shumalia et al., 2016). Excessive use of gutka, niswar, and manpuri contributes to the development of OSCC (Tanaka et al., 2011). Low socioeconomic status, poor education, and lack of knowledge about carcinogenic substances may contribute to this prevalence. Oral cancer sites vary widely, with buccal mucosa (41.7%) being the most involved site in oral squamous cell carcinoma (OSCC), followed by the tongue (20.0%). Studies from India and western countries have reported different oral cancer localizations, with the tongue being more frequent in western countries due to excessive alcohol use (Tanaka et al., 2011; Jerjes et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). In our population, excessive chewing tobacco may be the cause of buccal mucosa involvement. Many study reported the presence of oral lesion and leukoplakia as a prominent clinical feature in majority of OSCC patients (Napier et al., 2008). We also found that oral lesions and leukopalakia as the most prevalent clinical symptoms in OSCC patients, with oral lumps found in 17.8% of cases.\u003c/p\u003e \u003cp\u003eHere we demonstrated the role of AR in different histological grades of Oropharyngeal Squamous Cell Carcinoma (OSCC) tissues. We found that in different grades of OSCC mRNA is expressed at different concentrations. Higher expression was found in all three OSCC grades (PDSCC, WDSCC and, MDSCC) as compared to healthy individuals. This suggests that altered expression of AR might have a role in the growth and progression of OSCC. Studies have highlighted the importance of AR expression for the growth and progression of OSCC cells, with higher expression levels observed in premalignant and malignant tissue compared to normal mucosa tissue (Wu et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In prostate and breast cancer, the expression of AR has been well discussed, but its role in OSCC remains unclear. From previous studies it was found that AR expression is detected in 16% of OSCC cases, with significant differences between genders (Marocchio et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Mohamed et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In laryngeal carcinoma, AR expression is 68.3%, and in juvenile nasopharyngeal fibroma, it is highly expressed in tumor fibroblast cells (Chen et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Hagen et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1994\u003c/span\u003e). Fard et al. reported that in gastric cancer, AR is overexpressed in 57 patients at transcriptional level (Fard et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and in renal carcinoma, it is significantly higher in patients with pathological stage 2 tumors documented by Ha et al. (Ha et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). In contrast to our findings, some studies have reported the opposite role of AR, such as its antitumor role in ER-positive breast cancer stated by Hickey et al. and its low expression in OSCC tissue compared to normal control tissue (Colella et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe human androgen gene, encoding a full-length 110kda protein, has multiple shortened isoforms lacking functional domains. These are called splice variants (AR-Vs) (Cao et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). These splice variants (ARs-Vs) have the ability to execute transcription, resulting in an uncontrollable tumor growth, even when the androgen signaling is absent (Chan et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Watson et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Studies have reported the presence of multiple splice variants of AR with ARVs mutated or truncated at the ligand binding domain.in prostate and breast cancers. The most detected and reviewed AR-Vs is AR-V7 (Watson et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). A direct relation exist between AR expression and serum testosterone levels. Thus, an enhanced testosterone levels can modify AR expression and stimulate cell proliferation in advanced stages of OSCC. We also found a strong correlation of serum testosterone between healthy oral tissues and histologically graded OSCC tissues. In poor and moderately differentiated tissue the testosterone levels were higher as compared to well differentiated OSCC. It implies that as the cancer progressed the testosterone levels rose. This is the first study to simultaneously assess serum testosterone levels in OSCC, as well as expression of AR mRNA and protein. Hence, patients with oral squamous cell carcinoma may have serum testosterone levels that can serve as a potential biomarker for OSCC.\u003c/p\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eEthical consideration\u003c/h2\u003e \u003cp\u003e The research project was presented in front of Institutional Review Board (IRB) of DUHS and the ethical approval for the study design, experimentation and ethical matters was granted by the board (REF IRB-1535/DUHS/Approval/2020).\u003c/p\u003e \u003c/div\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eTo the best of our knowledge, no literature have been found that investigated the expression of AR mRNA and protein levels and correlation of serum testosterone levels with the expression of AR in OSCC patients. We first time investigated the expression of androgen receptor (AR) mRNA and protein levels and found that it is overexpressed in different histological grades of OSCC as compared to healthy individuals. The study also found a significant correlation between serum testosterone levels and AR expression in OSCC patients, and observed full length androgen receptor and its splice variant AR-V7 in poorly and moderately differentiated samples.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eNousheen Faizullah Khan: sample collection and processing, all protocols included in methodology, compilation of results and prepared all the figures and statistical evaluation, initial drafting of manuscriptRafat Amin: Idea conceptualization, protocol optimization, supervision of the study, manuscript editing and proof readingNeha Baqai: sample collection and processing, protocol optimization, statistical evaluation, manuscript proof readingSyeda Noureen Iqbal: provision of samples, clinical validation of study subjects, manuscript revisionZeba Ahmed: provision of samples, clinical validation of study subjects, manuscript revisionTehseen Fatima: Idea conceptualization, protocol optimization, supervision of the study, manuscript writeup, editing and final proof reading\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbdulla, R., Adyanthaya, S., Kini, P., Mohanty, V., D\u0026apos;Souza, N., \u0026amp; Subbannayya, Y. 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(2010). Clinicopathological parameters, recurrence, locoregional and distant metastasis in 115 T1-T2 oral squamous cell carcinoma patients. Head \u0026amp; neck oncology, 2, 1-21.\u003c/li\u003e\n\u003cli\u003eJohn, R. M., Yadav, A., \u0026amp; Sinha, D. N. (2018). Smokeless tobacco taxation: lessons from Southeast Asia. Indian Journal of Medical Research, 148(1), 46-55.\u003c/li\u003e\n\u003cli\u003eJohnson, N. W., Jayasekara, P., \u0026amp; Amarasinghe, A. A. (2011). Squamous cell carcinoma and precursor lesions of the oral cavity: epidemiology and aetiology. Periodontol 2000, 57(1), 19-37. https://doi.org/10.1111/j.1600-0757.2011.00401.x\u003c/li\u003e\n\u003cli\u003eKim, J. W., Park, Y., Roh, J. L., Cho, K. J., Choi, S. H., Nam, S. Y., \u0026amp; Kim, S. Y. (2016). Prognostic value of glucosylceramide synthase and P-glycoprotein expression in oral cavity cancer. 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I., Ahmed, F., \u0026amp; Mohammad, H. (2016). Role of chewing habits and cigarette smoking in differentiation of oral squamous cell carcinoma. Pak. J. Med, 5(3), 32-37.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"OSCC, androgen, testosterone, androgen receptors, squamous, oral cancer.","lastPublishedDoi":"10.21203/rs.3.rs-5387823/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5387823/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eOral squamous cell carcinoma (OSCC) is a common malignant tumor, in the head and neck region with a high incidence and prevalence rate globally. Oral cancer is one of the most deadly and common cancers among men in Asian nations including Bangladesh, India, and Pakistan. Certain viral infections, alcohol, betel quid, areca nut, and tobacco use have all been linked to the development of OSCC. Although the role of hormones and their receptors have been explored in many types of cancers, in OSCC it is still poorly explored. Previous studies have reported that sex hormone androgens and androgen receptors (AR) are involved in growth and progression of breast and prostate cancers. However their role in OSCC remains controversial.\u003cbr\u003e\nThe study aims to understand the expression patterns of the androgen receptor at the transcriptional and translational levels in various grades of patients with OSCC. Further to that the expression of AR changes in response to testosterone was also compared in OSCC parients in relation to healthy population.\u003cbr\u003e\nAfter obtaining consent, the participants were divided into four categories: healthy, poorly differentiated (PD), moderately differentiated (MD), and well differentiated (WD) squamous cell carcinoma (SCC). By creating an appropriate questionnaire, the patient's history was acquired. Samples of blood and tissue were obtained and processed, for testosterone level measurement using ELISA and RNA and protein estimation using commercially available kits, respectively. Tissue samples were visualized using H\u0026amp;E staining. We found that PDSCC group has highest expression levels of AR mRNA, followed by MDSCC and WDSCC. AR expression and serum testosterone levels were found to have moderately significant association in all subjects, determined by Spearman’s correlation. All groups showed full length AR expression while the splice variant AR-V7 was detected exclusively in MDSCC and PDSCC. This is suggestive of AR-V7 role in in OSCC progression. Higher levels of serum testosterone in OSCC patients than in healthy people provide an evidence for the carcinogenic potential of AR in OSCC.\u003c/p\u003e","manuscriptTitle":"Investigating the expression patterns of Androgen Receptor in different grades of Oral Squamous Cell carcinoma ","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-12-04 00:30:29","doi":"10.21203/rs.3.rs-5387823/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"15003b1e-661a-4781-9dd8-c12d3151f8d2","owner":[],"postedDate":"December 4th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-12-20T12:54:08+00:00","versionOfRecord":[],"versionCreatedAt":"2024-12-04 00:30:29","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5387823","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5387823","identity":"rs-5387823","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}