Uterine fibroids are the most common benign tumors of the female genital tract. Steroid hormones, especially estradiol and progesterone, play an important role in the pathobiology of this frequent disease. Recent studies suggested that both expression levels and polymorphisms of estrogen receptor (ER) alpha and beta might affect development of uterine fibroids. In this study, we tested whether single nucleotide polymorphisms (SNPs) in the promoter of estrogen receptor beta gene (ESR2) are associated with susceptibility to uterine fibroids. For this purpose, we compared the frequency of three SNPs in the promoter region of ESR2 gene (rs2987983, rs3020450 and rs3020449) in 101 women with uterine fibroids and 102 healthy women serving as controls by means of allele-specific tetra-primer polymerase chain reaction (PCR). Regarding allele frequency, allele positivity, genotype and haplotype frequencies of these SNPs we did not observe any significant difference between healthy women and women with uterine fibroids. In conclusion, our data clearly suggest that the tested SNPs in the promotor region of human ESR2 gene are not associated with the development of uterine fibroids.