This study aims to evaluate the protective effect of Benzethonium Chloride (BZC) on- diethylnitrosamine (DEN)‑induced hepatocellular carcinoma. Thirty-two male albino rats were used (eight animal per group). The hepatocarcinogenesis was induced in male Wistar albino rats by DEN (20 mg/kg b.w, orally (five times per week for 6 weeks). The treatment group rats received a similar dosage as above then injected with benzethonium chloride (5 mg/ kg b.w i.p.) for 4 weeks. Blood samples for serum parameters were collected or determination of  biochemical markers such as alanine amino transaminase (ALT), aspartate amino transaminase (AST), alkaline phosphatase (ALP), and total bilirubin, albumin and total protein. Also, liver tissues were taken for histopathological investigation. The obtained result revealed that treatment with benzethonium chloride significantly reduced DEN-induced elevation of ALT, AST and ALP activities and total bilirubin concentration. However, significant increase in the serum albumin and total protein concentration were observed in benzethonium chloride-treated rats as compared with DEN‑treated  group. The obtained results suggest that  benzethonium chloride possesses chemopreventive activity against DEN‑induced liver cancer.