In recent years, nanoscience has emerged as a prominent research field, focusing on exploring the therapeutic properties of herbal medicine. This research presents the green synthesis of Ag-doped ZnO nanoparticles using the fruit extract of Sophora pachycarpa (S. pachycarpa@Ag-doped ZnO NPs). Various characterization methods, including XRD, EDAX, TEM, and FT-IR, were employed to identify the nanoparticles. The XRD technique confirmed the crystallinity of S. pachycarpa@Ag-doped ZnO NPs, with an average size of 47nm. In this investigation, we explored the potential toxicity of S. pachycarpa fruit extract and S. pachycarpa@Ag-doped ZnO NPs on U266B1 cells. The cells were treated for 24 and 72hours with varying concentrations (0.5-1mg/ml) of S. pachycarpa and S. pachycarpa@Ag-doped ZnO NPs. The percentage of cell viability was determined using the AlamarBlue Assay. Additionally, the expression level of the P53 gene was evaluated by Real-Time PCR, with the β-actin gene serving as an internal reference gene for comparison. Viability results demonstrated that approximately 92% and 62% of cells remained alive after treatment with 1mg/ml dose of S. pachycarpa for 24 and 72hours, respectively, compared to the control. In the case of cells treated with S. pachycarpa@Ag-doped ZnO NPs, an IC50 of 585.664µg/ml was acquired after 24h of treatment. The expression level of P53 in cells treated with S. pachycarpa@Ag-doped ZnO NPs and S. pachycarpa significantly increased in a concentration-dependent manner, reaching 10380 and 1.62 fold expressions in 2mg/ml of S. pachycarpa@Ag-doped ZnO NPs and S. pachycarpa, respectively. Based on the obtained results, it can be concluded that S. pachycarpa@Ag-doped ZnO NPs can be considered a new avenue for Multiple myeloma (MM) treatment. However, further studies are necessary to identify more detailed aspects of the drug's mechanism of action, such as investigating the signaling pathway and protein level via western blotting.