Caffeine-boosted silver nanoparticles target breast cancer cells by triggering oxidative stress, inflammation, and apoptotic pathways

Abstract

Breast cancer (BC) constitutes a major global health concern and is the second foremost cause of cancer-related mortality among women worldwide. This research investigated the anticancer effectiveness of caffeine-conjugated silver nanoparticles (Caf-AgNPs) against MDA-MB-231 breast cancer cells, utilizing fluorouracil (5-FU) as a reference antitumor drug. The study illustrated that the strategic conjugation of caffeine with AgNPs substantially improved the therapeutic efficacy against breast cancer cell lines and simultaneously attenuated cytotoxicity in normal mouse liver (NBL) cells. Caf-AgNPs significantly increased ROS, malondialdehyde, COX-2, IL-1β, and TNF-α level in BC cells, which was accompanied by a decrease in glutathione levels. The increased levels of cytosolic cytochrome c, caspase-3, and Bax proteins, as well as a significant decrease in Bcl-2 expression and Bcl-2/Bax ratio, were indicative of the significant pro-apoptotic effects of Caf-AgNPs in MDA-MB-231 cells. Cancer cells subjected to Caf-AgNPs demonstrated elevated lactate dehydrogenase (LDH) membrane leakage, signifying cellular membrane disruption. Cell cycle analysis revealed a substantial proportion of early and late stage apoptosis in cancer cells exposed to Caf-AgNPs, accompanied by a notable downregulation of cyclin D1 and cyclin-dependent kinase 2 (CDK2) mRNA expression. Caf-AgNPs utilize several mechanisms for cellular destruction, including cell cycle arrest, oxidative stress induction, modulation of the inflammatory response, and mitochondrial apoptosis. Caf-AgNPs offer a promising and complex strategy for breast cancer intervention. © 2025 American Pharmacists Association