Cancer poses a significant challenge in the medical field, requiring thorough investigation into its mechanisms and the development of effective treatments. Recently, there has been increasing interest in integrating drugs with metal nanoparticles, which are notable for their unique size and physicochemical properties, aiming to enhance anticancer efficacy. Silver nanoparticles (AgNPs), especially those obtained through environmentally friendly methods known as green synthesis, have garnered attention. Royal jelly (RJ), a substance produced by bees recognized for its antioxidant, anti-inflammatory, and antibacterial properties, is particularly interesting. This study focuses on the green synthesis of AgNPs using royal jelly and its bioactivity against cancer cells. It provides a detailed characterization of the nanoparticles and examines their effects on cancer cells, specifically Hela cervical cancer and A549 lung cancer cell lines. The results highlight the cytotoxicity induced by AgNPs in HeLa and A549 cells, which is mediated through apoptosis via the PI3K/AKT signaling pathway. Our research findings demonstrate that one of the mechanisms underlying cell death involves increased concentration of ROS/RNS and downregulation of the VEGFa/MMP-2/COX-2 pathway. This study is among the few to elucidate the mechanism of the anticancer effects of nanoparticles synthesized through this method. Overall, our research contributes to the ongoing exploration of cancer biology and offers insights into potential therapeutic approaches by harnessing the capabilities of green-synthesized nanoparticles.