Monoterpene Sabinene Suppresses Hepatocarcinoma by Regulating the AKT/mTOR and Bcl‐2/Bax Signaling Pathways: An In Vivo and In Vitro Analysis

Essential oils (EOs) are aromatic compounds derived from plants, recognized for their diverse pharmacological properties that enable various therapeutic applications. Sabinene is a bicyclic monoterpene found in essential oils from plants such as Piper nigrum, Myristica fragrans, Origanum majorana and citrus fruits. It possesses pharmacological properties, can modulate immune responses, and may exhibit anticancer activity by inhibiting tumor cell proliferation and inducing apoptosis. We aimed to assess the anticancer efficacy of sabinene against hepatocarcinoma cells through both animal and in vitro studies. Male Wistar rats were induced with hepatocarcinoma via diethylnitrosamine (DEN) treatment, and the efficacy of sabinene was assessed in these rats, compared with the anti-inflammatory drug silymarin. We measured the loss in body and liver weight and recorded the incidence of tumors in the experimental rats. The renal and liver enzyme profiles were assessed to analyze the ameliorative efficacy of sabinene in the hepatocarcinoma condition. Tumor biomarkers such as AFP, CEA, and 8-OHdG were quantified to confirm tumor incidence due to DEN and the anticancer potency of sabinene. Antioxidant levels and interleukin concentrations were assessed to determine the antioxidant and anti-inflammatory effects of sabinene. The apoptotic efficacy of sabinene against hepatic carcinoma progression was measured by quantifying apoptotic, AKT, and mTOR proteins in the experimental animals. Liver histopathological assessment was done to analyze the anticancer efficacy of sabinene. For in vitro analysis, HepG2 and HL7702 cell lines were utilized to assess the cytotoxic efficacy of sabinene against normal and cancerous hepatocytes. Intracellular staining and apoptotic protein quantification were conducted to analyze the potency of sabinene in elevating reactive oxygen species (ROS) and triggering apoptosis in HepG2 carcinoma cells. Our results indicate that sabinene treatment prevented tumor incidence and hepatic injury in DEN-treated rats. It significantly attenuated tumor biomarkers, elevated antioxidant status, and regulated interleukins, thereby preventing inflammation and cancer induction. Sabinene also triggered apoptosis and inhibited tumor progression in DEN-treated rats. The in vitro analysis data correlated with the animal studies; sabinene treatment effectively triggered apoptotic signaling by enhancing intracellular ROS levels. The MTT assay on sabinene-treated HL7702 cells confirmed its non-cytotoxic effect against normal hepatocytes. Overall, our findings indicate that sabinene proves to be a potent anticancer agent in both animal and in vitro models and may serve as an effective alternative to current anticancer drugs for treating hepatocarcinoma. Our findings suggest potential therapeutic applications of sabinene in hepatocarcinoma treatment, although further research is needed to fully elucidate its mechanisms and clinical efficacy.