The bio-green methods of synthesis nanoparticles (NPs) have advantages over chemo-physical procedures due to cost-effective and ecofriendly products. The goal of current investigation is biosynthesis of zinc oxide NPs (ZnO-NPs) and evaluation of their biological assessment. Water extract of Brassica napus pollen [rapeseed (RP)] prepared and used for the synthesis of ZnO-NPs and synthesised ZnO-NP characterised using ultraviolet-visible, X-ray diffraction, Fourier-transform infrared spectroscopy, field emission scanning electron microscope and transmission electron microscope. Antioxidant properties of ZnO-NPs, cytotoxic and pro-apoptotic potentials of NPs were also evaluated. The results showed that ZnO-NPs have a hexagonal shape with 26 nm size. ZnO-NPs synthesised in RP (RP/ZnO-NPs) exhibited the good antioxidant potential compared with the butylated hydroxyanisole as a positive control. These NPs showed the cytotoxic effects against breast cancer cells (M.D. Anderson-Metastasis Breast cancer (MDA-MB)) with IC 50 about 1, 6 and 6 μg/ml after 24, 48 and 72 h of exposure, respectively. RP/ZnO-NPs were found effective in increasing the expression of catalase enzyme, the enzyme involved in antioxidants properties of the cells. Bio-green synthesised RP/ZnO-NPs showed antioxidant and cytotoxic properties. The results of the present study support the advantages of using the bio-green procedure for the synthesis of NPs as an antioxidant and as anti-cancer agents.