The present study was conducted to investigate the prognostic significance of co-expression patterna of HER-2,IL-6, TNF-a and TGF-β1 in breast cancer, by correlating the number of markers with positive expression withclinicopathological characteristics indicative of tumor progression and overall survival. One hundred thirtyconsecutive patients with primary breast cancer were prospectively included and evaluated. Serum concentrationsof the above markers were measured by ELISA. Median split was used to subdivide patients with marker positiveor negative expression. The presence of ≥3 positive markers was independently associated with extended lymphnode (>3) involvement (aOR, 11.94, p=0.001) and lymphovascular invasion (aOR, 12.04, p=0.018), increasingthe prognostic significance of each marker considered separately. Additional prognostic information regardingsurvival was also provided; as the number of positive markers increased, a gradually reduction of survival timewas observed. In addition, patients with 4 positive markers had significantly shorter survival (25 vs 39 months,p=0.006) and a more than 4 fold increased risk of death (aHR, 4.35, p=0.003) compared to patients with 3 positivemarkers. Our findings suggest that the coexpression pattern of these four markers could be used clinically as auseful marker for tumor extension and outcome of breast cancer.