Background: Esophageal cancer is one of the main causes of cancer mortality in the world. Golestan province, in the northern part of Iran, has the highest esophageal cancer rate in the world. The north and south districts of Golestan province can be classified as low and high-risk areas for esophageal cancer. One of the potential risk factors for esophageal cancer in this population is a nutrient-deficient diet. Dietary antioxidant compounds such as selenium, vitamin E, vitamin A, and β-carotene are reactive oxygen species (ROC) scavengers that play a key role in cellular responses to oxidative stress and preventing DNA damage. This study aims to compare the serum levels of selenium, vitamin E, and vitamin A in healthy individuals in high and low-risk areas of esophageal cancer. Methods: This study is a population of 242 healthy individuals. Serum selenium levels were assessed by atomic absorption spectroscopy. Vitamin E and A were assessed by reversed-phase high-performance liquid chromatography. Results: Vitamin E levels of healthy individuals in high-risk areas were significantly lower than in low-risk areas, while there was no significant difference between the selenium and vitamin A levels of healthy individuals in high-risk areas and low-risk areas. Also, there was no significant difference between selenium, vitamin E, and vitamin A levels in urban and rural areas and men and women in Golestan province. Conclusion: High levels of selenium with lower levels of vitamin E, along with other risk factors, may be associated with esophageal squamous cell carcinoma in high-risk areas of Golestan province.