Chemotherapeutic Potential of a Tri-Herbal Formulation Against NMU-Induced Mammary Tumors via Histological Remodeling and Genoprotection

Background and Objective: Herbal bioactives from Nigella sativa (black cumin), Boswellia sacra (frankincense), and Carica papaya (papaya leaf) possess potent antioxidant and anti-inflammatory activities. This study evaluated the histopathological and genotoxic effects of a tri-herbal formulation combining these botanicals in N-nitrosomethylurea (NMU)-induced mammary carcinogenesis in mice. Materials and Methods: Fifty female mice were randomly divided into 5 groups (n = 10): Control, NMU+cisplatin (2 mg/kg/week), NMU+Tri-herbal low dose (100 mg/kg/day), NMU+tri-herbal mid dose (200 mg/kg/day), and NMU+Tri-herbal high dose (400 mg/kg/day). Mammary tumours were induced via intraperitoneal NMU injections (50 mg/kg) administered weekly for three weeks. One week after the final NMU dose, treatments were given orally for eight weeks. Body weight and tumour growth were monitored weekly. After euthanasia, mammary and visceral tissues were examined histologically (H&E), and DNA damage in blood and tumour cells was quantified using the alkaline comet assay. All quantitative data were analysed using one-way analysis of variance followed by Tukey’s post hoc test, and differences were considered statistically significant at p<0.05. Results: Tri-herbal-treated groups showed significant reductions in tumour volume and improved body weight relative to NMU controls (p<0.05). Histopathology revealed reduced cellular proliferation, increased apoptotic activity, and near-normal glandular restoration. Comet assay indices (% tail DNA, tail moment) indicated markedly decreased genotoxicity compared with NMU+cisplatin, suggesting enhanced genomic stability. Conclusion: The tri-herbal combination of N. sativa, B. sacra, and C. papaya exhibits robust chemopreventive potential against NMU-induced mammary tumours through antioxidative, anti-inflammatory, and DNA-protective mechanisms, supporting its promise as a complementary phytotherapeutic in breast cancer management.