ObjectiveIn the present study, we green synthesized ZnO NPs (zinc oxide nanoparticles) from Senna occidentalis leaf extract which were subsequently assessed for their cytotoxic and antioxidant activity on colon cancer SW480 cell line.Results Zinc oxide nanoparticles were characterized by using UV-Vis spectroscopy, X-ray diffractometer (XRD), Particle size analyzer (PSA), Fourier transform infrared spectroscopy (FTIR), Field emission scanning electron microscopy (FE-SEM), Energy-dispersive X-ray (EDS) and atomic force microscope (AFM) analysis. The PSA, XRD and AFM showed 20-50 nm size and nearly cuboidal and irregular shaped of the ZnO NPs. The synthesized ZnO nanoparticles were evaluated for their anticancer activity on human colon cancer cell line (SW480) by using MTT and neutral red uptake assay. The SW480 colon cancer cells were treated with various concentrations of ZnO NPs in the range of 20–100 µg/ml for 2 hrs. The result showed that ZnO NPs could reduce cell viability of SW480 cells up to 50% at the concentration of 100 µg/ml and induce membrane leakage in a concentration dependent manner.ConclusionThe anticancer activity of zinc oxide nanoparticles has showed that these can be used as effective anti-cancer agent against colon cancer cell lines (SW480).