Background: Uterine leiomyomas could be considered as benign tumor of human uterus smooth muscle with unknown etiology and pathophysiology.Furthermore, they are the most common indication of hysterectomy. The tumor suppressor gene p53 has been involved in various malignancies. Mutation in its promoter site may play a role in tumorigenesis of many malignancies including leiomyoma. Materials and Methods: For study of polymorphisms and allele frequency, 234 female patients with pathologically diagnosed uterine leiomyoma and 100 healthy blood donors as control group were assessed. DNAs were extracted from peripheral blood cells, amplified using polymerase chain reaction and restriction fragment length polymorphism (RFLP) technique was utilized for their analysis. Result: Proportions of A homozygote/heterozygote/G homozygote for SNP -250 A/G in leiomyoma group were 97.8%, 1.7%, and 0.4%, and in control group 97%, 3%, and 0%, respectively. In case of -216 T/C polymorphism, proportions of T homozygote/heterozygote/C homozygote in leiomyoma were 98%, 1.7%, and 0%, and in control samples 98%, 2%, and 0%, respectively. Genotype frequency of A homozygote/heterozygote/G homozygote for SNP-103 A/G was 97.9%, 1.7%, and 0.4% in leiomyoma group, and 98%, 2%, and 0% in control group, respectively. Proportions of A homozygote/heterozygote/G homozygote for SNP-33 A/G in leiomyoma group were 97.8%, 2.2%, and 0%, and 97%, 3%, and 0% in case samples, respectively. Discussion: Based on the present results in an Iranian female population, surprisingly there was no significant differences between leiomyoma cases and control samples regarding allele frequencies of p53 promoter polymorphism.Therefore, The p53 promoter polymorphism is not associated with the susceptibility of uterine leiomyomas in Iranian women.