Plant-mediated metallic nanoparticles have beenreported for a diversified range of applications in biological sciences. In the present study, we propose the Polianthes tuberosa flower as a reducing and stabilizing agent for the synthesis of silver nanoparticles (PTAgNPs). The PTAgNPs were exclusively characterized using UV–Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), atomic force microscopy, zeta potential, and transmission electron microscopy (TEM) studies. In a biological assay, we investigated the antibacterial and anticancer activity of silver nanoparticles in the A431 cell line. The PTAgNPs demonstrated a dose-dependent activity in E. coli and S. aureus, suggesting the bactericidal nature of AgNPs. The PTAgNPs exhibited dose-dependent toxicity in the A431 cell line, with an IC₅₀ of 54.56 µg/mL arresting cell growth at the S phase, as revealed by flow cytometry analysis. The COMET assay revealed 39.9% and 18.15 severities of DNA damage and tail length in the treated cell line, respectively. Fluorescence staining studies indicate that PTAgNPs cause reactive oxygen species (ROS) and trigger apoptosis. This research demonstrates that synthesized silver nanoparticles have a significant effect on inhibiting the growth of melanoma cells and other forms of skin cancer. The results show that these particles can cause apoptosis or cell death in malignant tumor cells. This suggests that they could be used to treat skin cancers without harming normal tissues.