Abstract: Breast cancer patients with positive epidermal growth factor receptor (EGFR) expression have significantly worse post‐relapse prognosis than patients with negative EGFR expression. Vinorelbine (NVB) is usually reserved as a salvage therapy after anthracyclines and taxanes in patients with breast cancer. To see whether EGFR expression has a predictive value in NVB‐mediated effect on human breast cancer cells, we examined 50 primary breast cancer samples. Of these, 42% were found to be NVB sensitive by ATP‐tumour chemosensitivity assay. Sensitivity was correlated with EGFR expression level (p = 0.001). To dynamically examine EGFR’s effect on NVB sensitivity in breast cancer cells, we used the real‐time cell electronic sensing system with EGFR‐positive and EGFR‐negative breast cancer cell lines, MCF‐7 and MDA‐MB‐435s, respectively. MCF‐7 is NVB sensitive, while MDA‐MB‐435 is NVB resistant. NVB‐induced cytotoxicity to MCF‐7 can be partly reversed with inhibitory anti‐EGFR antibody. NVB up‐regulated EGFR expression in MCF‐7 cells, which affects ERK1/2 phosphorylation. This cellular response mechanism may cause greater input to non‐lethally damaged cells. These data suggest that EGFR expression can be used as a prognostic factor for breast cancer sensitivity to NVB, which could help identify appropriate treatments for breast cancer.