Hepatocellular carcinoma (HCC), the major primary malignant tumor of the liver, is one of the most life-threatening human cancers in the world, resulting in almost one million deaths every year. Currently available treatment options cause serious adverse effects. Considering the limited treatment options and dismal prognosis for HCC, chemoprevention has been considered the best strategy to reduce its current morbidity and mortality. In this regard, recently naturally occurring polyphenols are receiving increased attention because of their promising efficacy in several cancer models. Aim: The current study was aimed at assessing the effect of punicalagin on PI3K/Akt signalling Mechanisms in HepG2 cells in vitro. Methods: The Human Liver cancer cells (HepG2) were obtained from National Centre for Cell Science, Pune, India and were grown in culture flasks, separately containing DMEM medium, respectively supplemented with 10% FBS under 5% CO2, 95% air at 37°C. Upon reaching confluence, the cells were trypsinized and passaged. After 80% confluency, HepG2 cells were treated with different concentrations of punicalagin (50, 75, 100 and 150µg/ml) and cell viability was assessed by MTT assay. Antioxidant enzymes such as super oxide dismutase (SOD), Catalase (CAT) were assessed by sandwich-ELISA methods. Gene expression analysis of PI3K, Akt, and cytochrome-C were analyzed by Real Time PCR analysis and the data were analysed using one-way-ANOVA. Results: Punicalagin treatment to HepG2 cells significantly (p<0.05) reduced proliferation of cells at the concentrations of 75, 100 and 150mg/ml. Antioxidant enzymes such as SOD and CAT were found to be improved in drug treated HepG2 cells. It also potentially downregulated mRNA expression of PI3K, Akt and Cytochrome-C in liver cancer cells (p<0.05). Conclusion: Our current study clearly indicates that punicalagin potentially reduced the growth of the HepG2 cells by modulating the expression of PI3K/Akt signalling molecules facilitating antioxidant enzymes. Hence, punicalagin could be considered as one of the naturally occurring polyphenols for the treatment of liver cancer.