Cancer is a deadly disease and ranks as the second leading cause of death globally, which predominantly arises as a result of continuous exposure of human beings to carcinogenic agents and environmental contaminants. Natural compounds can potentially treat up to 60% of cancer cases. Therefore, this study investigated the anticancer and antioxidant activities of pomegranate peel extracts, which were prepared using a novel hybrid ultrasound-microwave assisted extraction (HUME) method. The HUME demonstrated significant improvements in extraction yield and polyphenol contents compared to conventional methods. The anticancer activity of pomegranate peel extract (PPE) was evaluated against HeLa, colon, and HepG2 cell lines and in vivo colon cancer in albino mice. The PPE demonstrated potent antioxidant activity, with DPPH and ABTS radical scavenging capacities of 91% and 95%, respectively. This potent activity was attributed to its high levels of phenolic acids (181 mg GAE/g) and flavonoids (35 mg QE/g). The secondary metabolites of PPE were detected by MS/MS using negative and positive ionization modes; the most abundant detected compounds were 4-hydroxycoumarin, p-coumaric, and gallic acid. A 40 Albino mice were randomly assigned to four experimental groups: a control group, a pathogen-induced cancer group, a 5-fluorouracil-treated group, and a group administered pomegranate peel extract (PPE). Albino mice treated with the PPE showed significant (P<0.05) tumor volume reduction and tumor growth inhibition compared to the diseased group. Pomegranate peel extract (PPE) demonstrated in vivo anticancer activity against bacterial pathogen-induced colon cancer. This was evident by significantly (P<0.05) downregulated B-cell leukemia/lymphoma 2 (BCL2) and Hypoxia-inducible factor 1-alpha (HIF1-α) level in the serum by 50% and 30%, respectively, in the PPE-treated group compared to the infected control. Concomitantly, PPE treatment improved histopathological features in the colon tissue, where the glandular structures remained intact and normal histological structure. These findings suggest that pomegranate peel extracts obtained by HUME possess potent anticancer and antioxidant activities. Further research is warranted to elucidate the underlying mechanisms and explore their potential as promising therapeutic agents for cancer treatment.

To Cite This Article: Altarjami LR, 2025. Anticancer and antioxidant activities of polyphenolic pomegranate peel extracts obtained by a novel hybrid ultrasound-microwave method: In vitro and in vivo studies in albino mice with HeLa, colon, and HepG2 cancerous cell lines. Pak Vet J. http://dx.doi.org/10.29261/pakvetj/2025.154