Punica granatum, specifically the fruit, has a long ethno medical history and is a phytochemical reservoir of great medicinal value. The phytochemistry and pharmacological actions of all P. granatum components suggest a wide range of clinical applications. The aim of the present study is to investigate the anticancer potential of aqueous extract of P. granatum (AEPG) in experimental models. The chemical composition of the AEPG was assessed by HPLC-DAD. In vivo antitumor activity was assessed in sarcoma 180 bearing mice. To evaluate the toxicological aspects related to the AEPG treatment, hematological, biochemical, histopathological and morphological analyses of treated animals were performed. Gallic acid, punicalagin α, punicalagin β, and ellagic acid were identified as the major phytochemical compounds of the extract. AEPG and 5-fluorouracil (5-FU) induced significant inhibition of tumor growth when compared with saline (p < 0.05). The percentage of apoptotic cells was significantly increased in 5-FU (p < 0.01) and AEPG treated groups (p < 0.01). No significant difference was observed between 5-FU and the three doses of AEPG. 5-FU induced toxic effects, such as decrease of body weight, splenic atrophy, and leukopenia, but these effects were not found in AEPG treated groups. The results provide evidence that AEPG exhibits comparable antitumor effects as 5-FU in a murine model, likely the result of increased apoptotic rate, but with no remarkable side effects presented by 5-FU.