In order to overcome the limitations of conventional therapeutic systems in the treatment of cancer, nanoparticles (NPs) have been rapidly produced and developed as a separate treatment method for control of cancer. Synthesis of nanoparticles using plant-based materials (green synthesis), due to the easy and cost-effective synthesis, production of non-toxic, sustainable and environmentally friendly products, can be considered the most appropriate method for preparation of NPs. In this study, after synthesis of Bi 2 O 3 NPs using Ginger ( Zingiber officinale ) root (rhizome) extract, the synthesized NPs were characterized and their potential application as selective anticancer agents against HCT116 colorectal cancer cells was evaluated through regulation of PI3K/AKT/mTOR signaling pathway, whereas the human kidney (HK-2) cells were used as normal cells. FTIR analysis showed a band at 673 cm −1 attributed to Bi-O vibration with a fingerprint region at 1291 cm −1 demonstrating the attachment of the organic molecules to the synthesized Bi 2 O 3 NPs. UV–visible study showed a λ max of around 268 nm, whereas XRD analysis showed eight clear peaks, demonetizing the crystalline phase of synthesized Bi 2 O 3 NPs. TEM analysis showed that spherical-shaped Bi 2 O 3 NPs have a size range of 20–50 nm with a man size of around 35 nm. Finally, DLS analysis determined that Bi 2 O 3 NPs have a hydrodynamic size of about 71.19 nm (PDI of 0.179) and a zeta potential value of −44.39 mV, revealing the good colloidal stability of NPs. Cellular assays (MTT, LDH, flow cytometry, and RT-qPCR) showed that synthesized Bi 2 O 3 NPs selectively induced anticancer effects against HCT116 colorectal cancer cells through membrane leakage, generation of ROS, induction of apoptosis via dysregulation of Bax, Bcl-2 and caspase-3 at mRNA level mediated via regulation of PI3K/AKT/mTOR signaling pathway. In conclusion, it may be suggested that the presence study could provide useful information for the potential anticancer effects mediated by synthesized Bi 2 O 3 NPs in vitro, although further studies, including in vivo studies and clinical trials, are needed to support our findings.