Background: Cancer development is a sequential process as a result of various cellular adaptation events. Oral cancer (squamous cell carcinoma) is the most predominant variety of head and neck cancer. Betanin (BTN) is isolated from beetroot extracts and is a highly bioavailable antioxidant. BTN exerts a chemopreventive and cytotoxic activity on numerous cancer cells. However, precise identification of the molecules responsible for this tumor-inhibitory effect is pending. This study aimed to understand the molecular mechanisms underlying the chemopreventive effects of BTN in 7,12-dimethylbenz (a) anthracene (DMBA)-induced oral cancer in experimental hamsters. Materials and Methods: The interactions of BTN with antioxidant enzymes, lipid peroxidation, apoptosis, and inflammatory markers in the presence of DMBA were investigated in male golden Syrian hamsters. Results: Oral supplementation of BTN treatment (50 mg/kg BW) daily to oral tumor-bearing rats successfully prevented DMBA-induced oral carcinogenesis. Furthermore, BTN administration significantly prevented weight loss and reduced the tumor occurrence, burden, volume, and biochemical parameters such as TBARS, LOOH, SOD, CAT, GPx, GSH, and vitamins E and C. The histological analysis and expression pattern of molecular markers (increased apoptosis (caspase-3 and 9), proliferative markers (PCNA and Cyclin-D1), and inflammatory markers (TNF-α and COX-2) investigated in hamsters' buccal mucosa tissues revealed a significant anti-tumorigenic nature of BTN. Conclusion: The findings of this study show that BTN markedly reduces DMBA-induced oral cancer in hamsters.