Skin cancer is a prevalent type of cancer, accounting for 30% of all cancer cases globally. Resveratrol (RSV), an anticancer drug, is effective against skin cancer but faces limitations such as poor aqueous solubility, first-pass metabolism, and instability, which hinder its topical use. This study aimed to develop and optimize RSV-loaded invasomes for topical administration and evaluate their efficacy in vivo. The optimized RSV-loaded invasomes exhibited small particle size (208.7 ± 74 nm), low polydispersity index (0.3 ± 0.03), high entrapment efficiency (77.7 ± 6%), and negative zeta potential (−70.4 ± 10.9 mV). They showed an initial burst effect followed by controlled drug release over 24 hours. The RSV-loaded invasomal gel demonstrated the highest skin deposition percentage (65%) in ex vivo rat skin, the lowest IC50 (6.34 μg/mL) in squamous cancerous cells (SCCs), and the highest cellular uptake compared to other formulations. In vivo studies in Ehrlich-induced mice models revealed that the RSV-loaded invasomal gel exhibited the smallest tumor volume with no signs of organ toxicity, indicating its safety and efficacy in treating skin cancer. RT-PCR and ELISA tests showed upregulation of BAX and Caspase-3 gene levels and downregulation of NF-kB and BCL2 protein levels, respectively. This study is the first to develop RSV-loaded invasomal gel for topical skin cancer treatment, demonstrating its potential as a promising lipid-based nanosystem for topical RSV delivery, with high skin penetration ability and anticancer effects.Skin cancer is a prevalent type of cancer, accounting for 30% of all cancer cases globally. Resveratrol (RSV), an anticancer drug, is effective against skin cancer but faces limitations such as poor aqueous solubility, first-pass metabolism, and instability, which hinder its topical use. This study aimed to develop and optimize RSV-loaded invasomes for topical administration and evaluate their efficacy in vivo. The optimized RSV-loaded invasomes exhibited small particle size (208.7 ± 74 nm), low polydispersity index (0.3 ± 0.03), high entrapment efficiency (77.7 ± 6%), and negative zeta potential (−70.4 ± 10.9 mV). They showed an initial burst effect followed by controlled drug release over 24 hours. The RSV-loaded invasomal gel demonstrated the highest skin deposition percentage (65%) in ex vivo rat skin, the lowest IC50 (6.34 μg/mL) in squamous cancerous cells (SCCs), and the highest cellular uptake compared to other formulations. In vivo studies in Ehrlich-induced mice models revealed that the RSV-loaded invasomal gel exhibited the smallest tumor volume with no signs of organ toxicity, indicating its safety and efficacy in treating skin cancer. RT-PCR and ELISA tests showed upregulation of BAX and Caspase-3 gene levels and downregulation of NF-kB and BCL2 protein levels, respectively. This study is the first to develop RSV-loaded invasomal gel for topical skin cancer treatment, demonstrating its potential as a promising lipid-based nanosystem for topical RSV delivery, with high skin penetration ability and anticancer effects.