<p>Lycopene is the highest concentration of red, hydrophobic carotenoids in various fruits and vegetables. Because of low solubility and poor bioavailability, a delivery system must be utilized to have a prolonged target therapeutic site. The current research is focused on assessing the potential antioxidant and anticancer activity of Lycopene Cubosomal Nanoparticles against the HT-29 colon cancer cell line. The nanoformulation was ensured by various characterization methods, including FTIR, TEM, SEM, particle size, and zeta potential. The structure of Lycopene was energy-minimized and was docked into the ATP-binding pocket using SwissDock with AutoDock Vina as backend. DPPH and ABTS were used for the presentation of antioxidant activities. In addition, anticancer activity of the compounds was tested by MTT viability test. Cell cycle and annexin-V/PI double staining were also investigated by flow cytometry. Further, gene expression and protein content of different markers PI3K, AKT, mTOR, Caspase-3, and Bcl-2 were analyzed by qRT-PCR and ELISA methods. The results presented that Lycopene Cubosomal Nanoparticles are more active than Lycopene as regards antioxidant and anticancer activity. Additionally, PI3K, AKT, mTOR, and Bcl-2 gene expression and content were decreased, and the gene expression and content of Caspase-3 were increased in both treatments, particularly pointing towards Lycopene Cubosomal Nanoparticles. Highly soluble Lycopene Cubosomal Nanoparticles were revealed to be more active than weakly soluble Lycopene alone, in that activity was enhanced over weakly soluble Lycopene alone, and Lycopene Cubosomal Nanoparticles were found to be more active than Lycopene alone as an antioxidant and anticancer agent. </p>