The present paper explains the straightforward procurement of Ag NPs interceded through gelatin as an inherent stabilizing and reducing agent in moderate condition for the treatment of breast cancer by investigating the P53 and STAT3 signaling pathways. Moreover, The formation of Ag NPs reinstated by gelatin and constructional features have been investigated through UV-Vis, FT-IR, TEM, FE-SEM, EDX and elemental mapping methods. The proofs and available information imply that gelatin-capped Ag NPs silver lead to the smallest mean dimensions of 10-15 nm. The cells exposed to Ag NPs/Gelatin were analyzed through the MTT assay over a period of 48 h to assess their cytotoxic effects on HUVEC and MCF10 breast cancer cells. The cancer cell demonstrated a decrease in viability upon exposure to Ag NPs/Gelatin, yielding an IC50 value of 125 mu g/mL. An in-depth examination of the P53 and STAT3 signaling pathway indicated that Ag NPs/Gelatin influenced Bax, Bcl2, P53 and pSTAT3/STAT3 in MCF10 through the pathway modulation. The Ag NPs/Gelatin reduced the cancer cells colony formation, G0/G1 and G2/M phases in cell cycle flow cytometry analysis and increased the S phase, LDH release (%), apoptosis (%), and ROS (a.u.). Furthermore, the antioxidant assessment indicated that the IC50 value for Ag NPs/Gelatin about DPPH was 15 mu g/mL. Therefore, Ag NPs/Gelatin have the potential to act as an advantageous natural anti-cancer agent in breast cancer treatment.