DIA/SWATH-Mass Spectrometry Revealing Melanoma Cell Proteome Transformations with Silver Nanoparticles: An Innovative Comparative Study

Melanoma is an aggressive cancer with increasing incidence and high mortality, driven by chemotherapy resistance and molecular dysregulation. Nanotechnology, specifically silver nanoparticles (AgNPs), shows promise as a treatment due to their ability to cause oxidative stress and apoptosis in cancer cells. However, traditional colloidal AgNPs lack selectivity, causing significant harm to normal cells. In this study, we present a green synthesis of AgNPs using plant extracts, which offers an eco-friendly alternative with a more targeted antitumor effect than traditional colloidal AgNPs. Using label-free DIA/SWATH MS quantitative proteomics, we compared the antitumor effects of AgNPs synthesized via traditional and green methods on A375 melanoma cells at 24 and 48 hours. Green AgNP exposure selectively reduced melanoma cell viability while sparing healthy keratinocytes (HaCaT), unlike Colloidal AgNPs. Proteomic analysis revealed that Green AgNPs exposure significantly decreased a panel of oncogenes and increasing carbohydrate metabolism, and dysregulating copper homeostasis. This is the first study to report the effects of differently synthesized AgNPs on melanoma cells using an integrated proteomic approach. It emphasizes the potential of green AgNPs in cancer therapy at a molecular level.