This study aimed to investigate the antitumor-promoting effects of total ethanolic extract from Perilla leaf (PLE) in a two-stage chemical carcinogenesis protocol in mice. Quantitative analysis active compounds in PLE were performed by thin layer chromatography with normal phase of silica gel F254 and HPLC-PDA with reversed phase of C18 column at UV 300 nm. The cutaneous tumors were initiated by a single application by 7,12-dimethylbenz[a]anthracene (DMBA) on the dorsal shaved skin, then promoted by repeated applications of croton oil during 20 weeks. PLE was topically applied daily until the end of the experiment. The results observed from thin layer chromatogram highlighted the presence of rosmarinic acid and luteolin in PLE. By means of HPLC-PDA analysis, the total content of rosmarinic acid and luteolin were found in amounts of 1.178 ± 0.011 % (w/w) and 0.105 ± 0.001 % (w/w), respectively. The PLE-treated group prolonged the latency period of tumor appearance up to 2 weeks and also significantly reduced tumor volume in dorsal skin of mice as compared to untreated group. In conclusion, the total ethanolic extract from leaves of Perilla frutescens demonstrated promising antitumor-promotion activity in mouse skin.