In vitro analysis of anti-cancer effects of sorafenib and parasetamol combination on hepatocellular carcinoma cells

Liver cancer ranks among the cancer types with the highest mortality rates worldwide, with hepatocellular carcinoma (HCC) being the most common form of primary liver cancer. Due to the limited efficacy of Sorafenib, a drug used in the treatment of HCC, and the development of drug resistance, the investigation of new combination therapies has become essential. Paracetamol, a widely used analgesic and antipyretic agent, is known to exhibit hepatotoxicity at high doses and has been reported to have potential anticancer effects. In this study, the cytotoxic, genotoxic, and apoptotic effects of the combination of Sorafenib and Paracetamol on the human liver cancer cell line HepG2 were evaluated in vitro. Cells were treated with various concentrations of Sorafenib and Paracetamol, individually and in combination, for 24 and 48 hours. Cell viability was assessed by MTT assay, and the LD₅₀ values were determined as 20 µM for Sorafenib and 5 mM for Paracetamol. DNA damage was analyzed using the Comet assay after 48-hour Sorafenib treatment, with the highest DNA damage observed at 20 µM Sorafenib (25.33 ± 4.16). A similar level of DNA damage (23.33 ± 1.15) was also detected following the combination of 20 µM Sorafenib and 5 mM Paracetamol for 48 hours. Additionally, gene expressions of BAX, BCL-2, CASPASE-3, and TNF-Α were analyzed by qRT-PCR, and protein levels were measured using the ELISA method. Results indicated that the combination treatment increased the expression of BAX and CASPASE-3, decreased BCL-2 levels, and elevated TNF-Α levels. These findings suggest that the combination of Sorafenib and Paracetamol exerts potent cytotoxic and genotoxic effects on HepG2 cells in a dose- and time-dependent manner, activates apoptotic pathways, and may possess synergistic anticancer potential. These data highlight the potential of combination strategies in HCC treatment and provide a significant foundation for further preclinical studies.