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Objectives: Estrogen is one of the important factors in the progression of breast cancer. Cytochrome P450 Superfamily (CYPs) plays an important role in the metabolism of estrogen. The CYP1A1 gene has been shown to be involved in the metabolism of estradiol. Several studies have reported a significant association between CYP1A1 polymorphisms and breast cancer. Methods:In the present work, the association between CYP1A1 polymorphism (Thr461Asn) and breast cancer in Guilan province was investigated. The blood samples were obtained from 120 patients and 80 controls. After DNA extraction, specific primers were adapted to amplify the specific fragment using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) and the results were examined through gel-electrophoresis. The product of amplification was digested with restriction enzyme BsmA1.Results: Genotype distributions of the CYP1A1 gene showed no significant difference between patients and controls (P = 0.12, Allele A in patients 0.16, in controls 0.10, Allele C in patients 0.84, and controls 0.9, respectively). Conclusions:The results from this study suggest that the Thr461Asn polymorphism of CYP1A1 gene may not be associated with the risk of breast cancer in the population of Guilan province. Further studies are needed to confirm the role of CYP1A1 gene in breast cancer.