Silver nanoparticles (AgNPs) were successfully synthesized using silver nitrate via the biological route using the culture filtrate of
Ganoderma enigmaticum
as well as
Trametes ljubarskyi
white rot fungi materials at room temperature. The proposed synthetic technique was applied for the first time for AgNPs preparation via the biological route through a low-cost pathway, which considered as an adequate direction of preparation compared to the commercial methods. This study reports the in vitro cytotoxic effect of biologically synthesized AgNPs in disposing of the human lung cancer cell line (A549) and human breast cancer cell (MCF-7) by using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. In addition, the viability of the tested cell lines was tested after treatment for 24 h in the presence of the prepared nanoparticles. The obtained results indicated the reduced viability of cancer cell lines with improving concentrations of AgNPs (40–120 μg/mL) at 24 h. Furthermore, at 120 μg/mL concentration, the fungal nanoparticles showed substantial cytotoxic effects toward the treated cells. Consequently, the results designated that the biologically synthesized silver nanoparticles have effective behavior for treating A549 and
MCF-7
cancer cells from the laboratory experiment approach; however, additional studies are required to validate these results in vivo models as anticancer agents depending on their cytotoxic activity.