Oxidative Stress, Inflammatory Dysregulation, and TGF-α-Mediated Tumor Promotion in Oral Squamous Epithelial Cells in individuals with Gutkha addiction

Abstract

Gutkha is widely consumed in South Asia. It is a smokeless tobacco containing areca nut and tobacco, and is associated with oral submucous fibrosis (OSF) and oral cancer due to the induction of inflammation and oxidative stress. This case-control study examined the effects of chronic gutkha chewing on oxidative stress markers, lipid profile, and inflammatory/tumor markers in male subjects aged 15–60 years from Sialkot, Punjab, Pakistan. Seventy habitual gutkha users (chewing ≥4–5 times/day for ≥10 years, with oral lesions) were compared to 30 age-matched healthy non-tobacco-using controls. Statistical analysis was done using R software (version 4.5.1; R version 4.5.1 (2025-06-13 ucrt) -- "Great Square Root"). Group comparisons were conducted using the independent two-sample t-test. Results showed a significant reduction in antioxidant enzyme activities in gutkha users: catalase (U/L), superoxide dismutase (U/L), reduced glutathione (µmol/L), and nitric oxide production (nmol/mg). Lipid profile (mg/dl) showed dyslipidemia with increased total cholesterol, triglycerides, and LDL-cholesterol. Inflammatory and tumor markers were markedly elevated: interleukin-12 (pg/ml) and transforming growth factor-α (pg/ml). The present findings indicate that chronic gutkha use induces oxidative stress, dysregulates antioxidant defenses, promotes dyslipidemia, and elevates pro-inflammatory and growth factor markers, potentially contributing to oral premalignancy and cancer progression. It is concluded that there is a novel association of elevated TGF-α and IL-12 with gutkha chewing in this population.