Resveratrol (RSV)-loaded invasome gel is a promising nanoformulation for the treatment of skin cancer. The study aimed to develop and optimize RSV-loaded invasomes for topical administration and assess their efficacy in vivo. The optimized RSV-loaded invasomes had a particle size of 208.7 ± 74 nm, a polydispersity index (PDI) of 0.3 ± 0.03, high entrapment efficiency (77.7 ± 6%), and a negative zeta potential (-70.4 ± 10.9 mV). They exhibited an initial burst effect followed by controlled drug release for 24 h. RSV-loaded invasomal gel showed the highest skin deposition percentage (65%) in ex vivo rat skin, the highest potency (lowest IC50 of 6.34 μg/mL), and the highest cellular uptake when tested on squamous cancerous cells (SCCs). The antitumor effect of topical RSV-loaded invasomes was evaluated in vivo in Ehrlich-induced mice models. The results revealed that RSV-loaded invasomal gel exhibited the smallest tumor volume with no signs of organ toxicity, indicating its safety in skin cancer treatment. Upregulation of BAX and Caspase-3 gene levels and downregulation of NF-κB and BCL2 protein levels were demonstrated using RT-PCR and ELISA tests. The study is the first to develop RSV-loaded invasomal gel for topical skin cancer treatment. Invasomes are considered promising lipid-based nanosystems for topical RSV delivery with high skin penetration ability and anticancer effect in the treatment of skin carcinoma.Resveratrol (RSV)-loaded invasome gel is a promising nanoformulation for the treatment of skin cancer. The study aimed to develop and optimize RSV-loaded invasomes for topical administration and assess their efficacy in vivo. The optimized RSV-loaded invasomes had a particle size of 208.7 ± 74 nm, a polydispersity index (PDI) of 0.3 ± 0.03, high entrapment efficiency (77.7 ± 6%), and a negative zeta potential (-70.4 ± 10.9 mV). They exhibited an initial burst effect followed by controlled drug release for 24 h. RSV-loaded invasomal gel showed the highest skin deposition percentage (65%) in ex vivo rat skin, the highest potency (lowest IC50 of 6.34 μg/mL), and the highest cellular uptake when tested on squamous cancerous cells (SCCs). The antitumor effect of topical RSV-loaded invasomes was evaluated in vivo in Ehrlich-induced mice models. The results revealed that RSV-loaded invasomal gel exhibited the smallest tumor volume with no signs of organ toxicity, indicating its safety in skin cancer treatment. Upregulation of BAX and Caspase-3 gene levels and downregulation of NF-κB and BCL2 protein levels were demonstrated using RT-PCR and ELISA tests. The study is the first to develop RSV-loaded invasomal gel for topical skin cancer treatment. Invasomes are considered promising lipid-based nanosystems for topical RSV delivery with high skin penetration ability and anticancer effect in the treatment of skin carcinoma.