Verbascum sinuatum and Nisin Probiotics Promote Apoptosis and Inhibit Metastasis by Up-regulation of p53 and Caspase 9 and Down-regulation ofMMP9 and SUR

Introduction: Breast cancer, the most common cancer in women, arises from genetic and epigenetic alterations that lead to aberrant gene function. Doxorubicin, a widely used chemotherapeutic agent, effectively inhibits cancer cell growth and proliferation but is associated with significant side effects. Natural substances, such as Verbascum sinuatum (V. sinuatum) and probiotics, may help reduce the required doses of chemotherapy drugs. In this study, we evaluated the relative expression of p53, Caspase 9 (CASP9), MMP9, and SUR in four treatment groups: V. sinuatum alone, Doxorubicin + V. sinuatum, Doxorubicin + Nisin, and V. sinuatum + Nisin. Methods: The MCF-7 breast cancer cell line was cultured, and the MTT assay was employed to assess cell viability. Gene expression analysis was performed using real-time PCR, evaluating the relative expression of CASP9, P53, SUR, and MMP9 in cells treated with V. sinuatum, Nisin, and Doxorubicin. Results: The IC50 of V. sinuatum in MCF-7 cells was 12.08 μg/mL. The greatest upregulation of P53 expression occurred in the Nisin + Doxorubicin group (7.95-fold, p < 0.0001), while CASP9 expression was highest in the Doxorubicin + V. sinuatum group (8.98-fold, p < 0.0001). The largest reduction in MMP9 expression was observed in the Nisin + Doxorubicin group (52.63%, p < 0.0001). Notably, SUR expression was undetectable in the Nisin and V. sinuatum groups. Conclusion: V. sinuatum extract and Nisin probiotic show potential as complementary treatments alongside chemotherapy, enhancing apoptosis and reducing metastasis-related gene expression in breast cancer cells.