Estrogen receptor alpha (ER-α) belong to the nuclear receptor superfamily of ligand-inducible transcription factors. It is involve in both mammary development and in breast carcinogenesis. Many polymorphisms suggested association with the incidence of breast cancer. The most widely studied Polymorphisms are PvuII and XbaI restriction sites, they lies within intron one of estrogen receptor alpha gene. This study was done to evaluate the relationship of two well- studied polymorphisms in the estrogen receptor alpha gene, (PvuII and XbaI), and the risk of breast cancer in Jordanian women. In our study, 100 blood samples from Jordanian women with breast cancer had been obtained and used for subsequent DNA extraction and genotyped for the PvuII and XbaI polymorphisms using Restriction Fragment Length Polymorphism (RFLPs) technique. Also, a 100 blood samples from normal Jordanian women had been used as control. For statistical analysis, SPSS computerized program to calculate P-value for polymorphisms and Hardy-Weinberg equation for genotype frequencies was used. There was no significant difference between breast cancer patients and healthy control individuals in the distribution of the PvuII genotypes. There was no significant difference between breast cancer patients and healthy control individuals in the distribution of the PvuII genotypes (P>0.05;P=0.27), no association between XbaI polymorphism and breast cancer risk (P>0.05;P=0.111). Also there was no significant association with haplotype analysis (P>0.05;P=0.07) except for AGTC haplotype (P≤0.05;P=0.043) and AATC haplotype (P≤0.05;P=0.027). This result suggests that PvuII and XbaI polymorphism doesn’t play a role in the susceptibility to breast cancer in Jordanian women