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The fight against cancer is one of the main challenges for medical research. Recently, nanotechnology has made significant progress, providing possibilities for developing innovative nanomaterials to overcome the common limitations of current therapies. In this context, silver nanoparticles (AgNPs) represent a promising nano-tool able to offer interesting applications for cancer research. Following this path, we combined the silver proprieties with <i>Artemisia</i> <i>arborescens</i> characteristics, producing novel nanoparticles called <i>Artemisia–AgNP</i>s. A “green” synthesis method was performed to produce <i>Artemisia–AgNP</i>s, using <i>Artemisia</i> <i>arborescens</i> extracts. This kind of photosynthesis is an eco-friendly, inexpensive, and fast approach. Moreover, the bioorganic molecules of plant extracts improved the biocompatibility and efficacy of <i>Artemisia–AgNP</i>s. The <i>Artemisia–AgNP</i>s were fully characterized and tested to compare their effects on various cancer cell lines, in particular HeLa and MCF-7. <i>Artemisia–AgNP</i>s treatment showed dose-dependent growth inhibition of cancer cells. Moreover, we evaluated their impact on the cell cycle, observing a G1 arrest mediated by <i>Artemisia–AgNP</i>s treatment. Using a clonogenic assay after treatment, we observed a complete lack of cell colonies, which demonstrated cell reproducibility death. To have a broader overview on gene expression impact, we performed RNA-sequencing, which demonstrated the potential of <i>Artemisia–AgNP</i>s as a suitable candidate tool in cancer research.