Green plant-based nanoparticles are becoming increasingly popular as a substitute for physical and chemical methods. A technique was used to synthesize selenium nanoparticles (SeNPs) using an ethanol extract of aerial parts of Adiantum capillus-veneris L. (ACV) as reducing agents for the H2SeO3 solution. The SeNPs were characterized using UV-vis, FTIR, AFM, SEM, XRD, EDX, and a zeta potential analyzer. The UV-Vis spectrum showed peaks at 269.0 nm and 311 nm. The FTIR spectrum of synthesized SeNPs confirms the existence of functional groups linked to bioactive molecules. The AFM results showed the lowest rate of nanosize reduction at 53.7 nm for nanosizes less than 100 nm. The SEM showed SeNPs with an appearance of small spherical with a diameter range of 27.91–12.28 nm. The XRD patterns of SeNPs synthesized by ACV extraction showed peaks at 28.48 and 58.76. The EDX spectrum showed elements such as Ca, Na, Mg, and K from the ACV leaf extract. The zeta potential of ACV SeNPs was 20.99 mV with a negative charge. The biological testing of SeNPs showed that they have antioxidant properties using the DPPH test, which was notably different from vitamin C. The in vitro cytotoxicity of SeNPs was investigated using the MTT assay. The results showed a non-toxic effect on the normal human fibroblast cell line (NHF) and anticancer activity against the breast cancer cell line (MCF-7).