In this investigation, the anticancer potentiality and biological mechanism of gold nanoparticles (AuNPs) was studied in SUDHL-4 cell line. Metallic AuNPs were prepared and stabilized with ethanol clove (Syzygium aromaticum) extract. The green synthesis of AuNPs was characterized and evaluated by UV-Visible Spectroscopic, X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Transmission electron microscopy (TEM), Dynamic Light Scattering (DLS) and biological activities using various biochemical assays. Green synthesis of AuNPs was confirmed by instrument method. The TEM images show polydisperse, mostly spherical AuNPs particles of 12 -20 nm. AuNPs were decreased the growth and viability of SU-DHL-4 cell line and increased the apoptosis. The treatments of SU-DHL-4 cells with AuNPs resulted in a moderate considerably increase in Reactive oxygen species (ROS) production. We measured apoptosis by Annexin-V/propidium iodide (PI) in the existence and nonexistence of the antioxidant N-acetyl-Lcysteine (NAC), the glutathione-depleting agent buthionine sulfoximine (BSO), or caspase inhibitors to determine the mechanism of cell death. AuNPs are unique potential anticancer agents that cause ROS-dependent apoptosis in SUDHL-4 cell line which was improved by depletion of glutathione (GHS) and inhibited by N-acetyl-L-cysteine on Z-VAD-FMK.