SiO2, commonly referred to as silica, represents a versatile material utilized across various domains including biomedical, construction, food, energy, and cosmetic sectors. This study is anticipated to offer insights into the synthesis of SiO2-NP from industrial hemp waste, thus contributing to sustainability efforts and waste management systems. Within this investigation, SiO2-NP were biogenically synthesized utilizing waste leaves from industrial hemp, a sustainable plant source. The biogenically synthesized SiO2-NP underwent characterization via several techniques including UV–visible absorption spectroscopy (UV–Vis), X-ray diffraction (XRD), Energy Dispersion X-ray Spectrometer (EDX), and Scanning Electron Microscopy (SEM) analysis. The UV–Vis spectrum indicated an absorption peak of 360 nm for SiO2-NP. XRD analysis determined an average particle size of 28 nm. SEM analysis revealed spherical morphology of the synthesized SiO2-NP. Additionally, EDX analysis unveiled a composition consisting of 50.6 % O and 27.2 % Si. SiO2-NP exhibited antibacterial activity against both gram-positive and gram-negative bacteria strains. According to anticancer activity analyses, IC50 doses of SiO2-NP in MCF7 and MDA-MB-231 breast cancer cells were found to be 146.73 ± 7.54 and 54.93 ± 5.54, respectively, for 48 h. After treating the cells with these doses, an increase in the expression of apoptosis-related genes was detected. The SiO2-NP has an IC50 value with 2.69 ± 0.42 mg/mL against AChE enzyme activity.