ABSTRACT Background: Toll-like receptors (TLRs) have generated an extraordinary amount of interest in cancer research in the last decade. More recently, TLR-mediated signaling pathways have been shown to support tumor cell growth in vitro and in vivo. Aim: Estimating the concentrations of TLR6 and TLR4 in serum and breast tissue of women with breast tumors. Materials and Methods: Blood and breast tissue were collected from 50 women (14–60 years) undergoing breast surgery at AL-Hilla Teaching Hospital and Al-Sadiq Hospital in Babylon Province. Histological confirmation of breast diseases, whether benign or malignant tumors, was done by the histological laboratory in the hospital. Twenty blood samples were collected from healthy women as controls. TLR4 and TLR6 were determined by the enzyme-linked immunosorbent assay in the serum of patients and controls as well as they were also determined in supernatant of cells in patients’ breast tissues. Results: The mean of TLR4 concentration was nonsignificantly increased in the serum of patients 4.58 ± 1.60 pg/ml than controls 4.18 ± 0.72 pg/ml (P = 0.28). The systemic mean of TLR6 in the serum of the patient was 2.11 pg/ml, while in control was 1.58 pg/ml with significant differences (P = 0.01). TLR4 concentrations also appeared systemically higher than local in different diseases. TLR6 concentrations were increased in local (tissue) than systemic with different disease types. There was a significant positive correlation between the concentration of TLR4 and TLR6 in the serum (systemic) of women with breast tumors (r = 0.353; P = 0.04). Conclusions: The systemic mean of TLR6 in the serum of the patient was 2.11 pg/ml, while in control was 1.58 pg/ml with significant differences (P = 0.01).