Cytotoxicity and Mitochondrial‑Mediated Apoptosis Induced by Fenugreek Seed Oil in Human Hepatocellular Carcinoma Cells via Reactive Oxygen Species Generation

Objective: The present study was aimed to investigate the anticancer activity of Fenugreek seed oil (FSO) against human hepatocellular carcinoma (HepG2) cell line. Materials and Methods: The HepG2 cells were exposed to 10–1000 µg/ml of FSO for 24 h to assess the cytotoxicity using 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyl tetrazolium bromide test, neutral red uptake assay, and morphological changes. The cytotoxic concentrations of FSO were used to study the induction in reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) levels. Further, the expression of pro‑and antiapoptotic genes (p53, caspase‑3, caspase‑9, Bax and Bcl‑2) were studied. Results: A concentration‑dependent decrease in cell viability of HepG2 cells was observed after FSO exposure. Our results showed that FSO decreased cell viability of HepG2 cells in a concentration‑dependent manner. FSO was also found to increase the ROS generation and decrease the MMP level in HepG2 cells. Our gene expression study indicates upregulation in apoptotic marker genes and downregulation in antiapoptotic gene. Conclusion: The results showed that FSO is capable to induce mitochondrial‑mediated apoptosis through ROS generation in HepG2 cells and could be a useful resource in developing effective remedies for the treatment of hepatocellular carcinoma.