In the present work, we synthesize Fe3O4/ZnO nanocomposite by sol-gel method using iron nitrate, ethylene glycol, zinc acetate dehydrate, and ammonium carbonate as precursors. Amorphous matrix and crystalline grains were formed in composite as well. The nanostructural properties of the nanocomposite were characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), transmission electron microscopy (TEM), vibrating sample magnetometer (VSM), and colloidal stability was checked out by Zeta potential. The biological effects of Fe3O4/ZnO nanocomposite were investigated by microculture tetrazolium test (MTT) against MCF-7 breast cancer cells as usage in medicine. Crystallite size, lattice strain, and dislocation density of the nanoparticles were estimated with Williamson-Hall method. The obtained results show that the highest death rate of Michigan Cancer Foundation-7 (MCF-7) cancer cells was for Fe3O4/ZnO nanocomposite with the molar ratio is 1:10 in the lower concentration (49 μ g/ml). Therefore, these nanocomposites due to their small size, colloidal stability, magnetic properties, and nontoxicity can be a good candidate as option in medicine application.