Objectives: Members of epidermal growth factor receptor (EGFR) family and cyclin D1 are frequently overexpressed in breast cancer. However, studies that systematically examine protein expression and gene amplification of HER-2, EGFR and cyclin D1 in breast cancer and correlate it with overall survival in the same group of patients are missing; hence results of the different studies are frequently opposite. Emerging evidences suggest the existence of a new mode of EGFR signaling pathway in which activated EGFR undergoes nuclear translocalization and subsequently regulates gene expression and potentially mediates other cellular processes. Therefore the aim of this study is to examine protein expression and gene amplification of HER-2, membrane EGFR (mEGFR), nuclear EGFR (nEGFR), cyclin D1 and chromosome 17 status in invasive ductal breast carcinoma and correlate findings with pathohistological parameters, menopausal status and overall survival. Material and Methods: Tissue microarrays (TMA) were built from the 118 archival, formalin fixed paraffin embedded invasive ductal carcinoma. Immunohistochemistry was performed for HER-2, mEGFR, nEGFR and cyclin D1 according to manufactures instructions. Gene evaluation of HER-2, EGFR, cyclin D1 and chromosome 17 status were performed by fluorescent in situ hybridization (FISH) using Abbott, Vysis probes (Downers Grove, IL, USA). Results: HER-2 overexpression was found in 23.27% tumors, corresponding gene amplification was detected in 18.10% samples while 20.68% cases had chromosome 17 polisomy. mEGFR protein overexpression and EGFR gene amplification were found in 1.8% of tumors. nEGFR expression was detected in 40.69% tumors with 11.50% having high nEGFR staining (nEGFR +++). Strong cyclin D1 expression and CCND1 amplification were found in 63.71% and 13.4% of the cases, respectively. Statistically significant correlation was found between immunohistochemistry and FISH analysis for HER-2, EGFR and cyclin D1. Univariate analysis for HER-2 gene amplification showed borderline statistical significance with shorter overall survival (OS) in whole cohort of patients (p=0.052) and statistically significant correlation with shorter OS in premenopausal group of patients (p= 0.0324). Strong cyclin D1 expression significantly correlated with better OS in pre-menopausal group of women (p= 0.0027) but there was no significant correlation in post-menopausal group or whole patient’s cohort although trend toward better survival has been noticed. Importantly, Kaplan-Meier survival analysis and log-rank test revealed a significant inverse correlation between high nEGFR and OS in whole cohort (p= 0.0002) as well in premenopausal group of patients (p= 4.43 x 10-6). Also, tumors with moderate/strong expression of cyclin D1 and nEGFR had significantly shorter cumulative survival comparing to tumors with moderate/strong cyclin D1 expression and negative/low nEGFR (p=0.00078). Multivariate analysis revealed nEGFR to be independent prognostic factors for OS and showed 3.4 times greater mortality risk for nEGFR+++ patients comparing to nEGFR negative patients (hazard ratio = 3.402; p=0.0026). Conclusion: Univariate analysis showed that patients with high nEGFR staining have statistically significant worse OS while multivariate analysis revealed that nEGFR represents independent prognostic factor. Also, our data indicate that in context of high nEGFR staining, increased expression of cyclin D1, actually contributes to the poor survival rate in invasive ductal breast cancer, but when expressed alone strong cyclin D1 expression represents a good prognostic marker. Our findings together with earlier reports suggest that nEGFR may play potential important role in the aggressive biology of breast cancer and subsequently can lead to worse clinical outcomes.