This study evaluated the hepatoprotective and anticancer potential of Ceratonia siliqua L. (Carob) bean extract (CSBE) against N-diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in Wistar rats, focusing on its modulation of biochemical, antioxidant, and molecular pathways, including the HOTAIR/miR-124/Notch1 axis. Fifty rats were divided into five groups: normal controls, DEN-induced HCC (80 mg/kg), HCC treated with 200 or 400 mg/kg CSBE, and normal rats receiving 400 mg/kg CSBE. Post-treatment assessments included serum liver function markers (albumin, total protein, bilirubin, CRP, ALT, AST, ALP), inflammatory cytokines (IL-6, IL-1β, IL-10, TNF-α), oxidative stress parameters (GPx, SOD, CAT, nitric oxide), and molecular analysis of HOTAIR, miR-124, Notch1, and Jagged1 expression in liver tissue, alongside p53 and caspase-3 immunostaining. DEN administration disrupted body/liver weights, elevated liver enzymes, impaired antioxidant defenses, and dysregulated the HOTAIR/miR-124/Notch1 axis, while CSBE—particularly at 400 mg/kg—dose-dependently restored hepatic function, reduced oxidative stress and inflammation, normalized gene expression, and improved histopathological architecture. These findings demonstrate CSBE’s efficacy in mitigating DEN-induced HCC through multifaceted mechanisms, highlighting its therapeutic potential.

KEY WORDS: Ceratonia siliqua L.; N-diethylnitrosamine; HOTAIR; Liver; Hepatocellular carcinoma.

GAO, X.; YING, X.; CHEN, D.; CHEN, Z.; LV, X. & WANG, J. Investigating the anticancer effects of Ceratonia siliqua L. (Carob) bean extract on hepatocellular carcinoma: insights into inflammatory, oxidative stress, and HOTAIR/miR-124/Notch1 pathways using biochemical, molecular, immunohistochemical, and histopathological methods. Int. J. Morphol., 44(2):789-802, 2026.