In this study, an unpretentious, non-toxic, and cost-effective dissolution casting method was utilized to synthesize a group of anticancer and biologically active hybrid nanocomposite materials containing biopolymer cellulose acetate. Pristine ZnO and Ag(0.01, 0.05, 0.1)/ZnO hybrid nanofillers based on variable Ag NP loadings were prepared via green procedures in the presence of gum arabic (GA). The chemical structures and the morphological features of the designed nanocomposite materials were investigated by PXRD, TEM, SEM, FTIR, TGA, and XPS characterization techniques. The characterization techniques confirmed the formation of CA@Ag(0.01, 0.05, 0.1)/ZnO hybrid nanocomposite materials with an average crystallite size of 15 nm. All investigated materials showed two degradation steps. The thermal stability of the fabricated samples was ranked in the following order: CA/ZnO < CA@Ag(0.01)/ZnO < CA@Ag(0.05)/ZnO = CA@Ag(0.1)/ZnO. Hence, the higher Ag doping level slightly enhanced the thermal stability. The developed nanocomposites were tested against six pathogens and were used as the target material to reduce the number of cancer cells. The presence of Ag NPs had a positive impact on the biological and the anticancer activities of the CA-reinforced Ag/ZnO composite materials. The CA@Ag(0.1)/ZnO hybrid nanocomposite membrane had the highest antimicrobial activity in comparison to the other fabricated materials. Furthermore, the developed CA@Ag(0.1)/ZnO hybrid nanocomposite material effectively induced cell death in breast cancer.