Chemotherapy, a gold standard for treating most of the cancers, involves drastic side-effects and multidrug resistance. An attractive alternative is development of combination therapy employing antimicrobial peptides with chemotherapeutic drugs. In vivo studies: Anti-cancer therapeutic efficacy of 5-fluororuacil (5-FU) in conjunction with nisin (50 mg/kg body weight) was evaluated against murine skin cancer, in terms of tumor biostatistics, histopathology, electron microscopy, infrared spectroscopy and transcriptional studies. In vitro studies: Dose and time dependent cytotoxicity of agents were assessed against A431 cell line using MIT assay, LDH assay and acridine orange/ethidium bromide dual staining. Significant percentage decrease(s) in mean tumor volume and tumor burden were observed in nisin+ 5-FU combination treated groups as compared to alone treated groups. Histoarchitecture of treated skins demonstrated restoration towards normal skin tissue (being highest in the combination group). Modulation of apoptotic, angiogenic and proliferative genes were observed in treated groups. IC 5 0 of combination was found to be 2 mu g/ml as compared to nisin alone (32 mu g/ml) and 5-FU alone (16 mu g/ml) with combination index of 0.188. Dual staining showed that rate of induction of apoptosis was higher in the combination group as compared to single agents. Nisin and 5-FU in combination were found to be synergistic both in vivo and in vitro. (C) 2019 Published by Elsevier Inc.