Background <p>Graviola emerged as a promising anticancer agent, with nanotechnology enhancing drugs’ therapeutic potential. The purpose of this work was to explore graviola extract and its nano-platform’s effects on tongue carcinoma (SCC154 cells) in vitro model.</p> Methods <p>Graviola leaves extract (GLE) was isolated, and its phenolic content was identified. Three nano-formulations (F1-F3) were optimized for GLE delivery, with F1 chosen for its optimal size and stability to synthesize graviola nanoparticles (GNPs). SCC154 cells were split into three groups: group Ι (untreated SCC154 cells), group ΙΙ: SCC154 cells + ethanolic GLE, and group ΙΙΙ: SCC154 cells + GLE encapsulated in a nano-void delivery system (GNPs). In vitro tests assessed cell viability via MTT assay, cell cycle, and apoptosis by flow cytometry, DNA damage using comet assay, and gene expression of the key molecular markers (PI3K, AKT, mTOR, and GSK-3β) by quantitative real-time polymerase chain reaction. Transmission electron microscopic examination of cells was also performed.</p> Results <p>GLE and GNPs reduced SCC154 cells’ proliferation compared to untreated cells, with GNPs showing significantly higher cytotoxicity. Both treatments also induced apoptosis, arrested the cell cycle, and caused DNA damage with a significant pronounced effect in the GNPs-treated group. Gene expression analysis revealed a substantial decline in PI3K, AKT, mTOR, and GSK-3β in both treated groups relative to the control group, with a significant downregulation in the GNPs-treated group. Ultrastructural examination revealed severe destruction in tongue carcinoma cells of both treated groups, with substantial damage in the GNPs-treated group.</p> Conclusion <p>GNPs showed a better impact than GLE in tongue carcinoma therapy, causing cytotoxicity and apoptosis, potentially through the PI3K/AKT/mTOR pathway.</p>