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

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and remains one of the leading causes of cancer-related deaths worldwide. It is frequently associated with chronic liver diseases, such as hepatitis B and C virus infections, cirrhosis, and excessive alcohol consumption. Although Sorafenib is the first FDA-approved targeted therapy for advanced HCC and has demonstrated significant efficacy in inhibiting tumor growth by targeting angiogenesis and cell proliferation pathways, its clinical effectiveness is often limited due to acquired drug resistance and adverse side effects. Therefore, combination therapies involving Sorafenib and other pharmacological agents have garnered attention as potential strategies to enhance therapeutic outcomes and overcome drug resistance. In this study, the in vitro anticancer effects of the combination of Sorafenib and Favipiravir a broad-spectrum antiviral agent known for its RNA polymerase inhibitory activity were investigated using the HepG2 human HCC cell line. The cells were treated with Sorafenib and Favipiravir both individually and in combination at predetermined doses and durations. Cell viability was evaluated using the MTT assay, while genotoxic effects were assessed through the Comet assay. Furthermore, quantitative real-time polymerase chain reaction (qRT-PCR) was performed to analyze changes in the expression levels of apoptosis-related genes (BAX, BCL-2, CASPASE-3, TNF-α), and protein-level changes were examined using ELISA. The results demonstrated that the Sorafenib and Favipiravir combination significantly reduced cell viability compared to monotherapies, indicating a synergistic cytotoxic effect. The combination also induced considerable DNA damage and significantly altered the expression levels of genes involved in apoptotic pathways, promoting programmed cell death. Comet assay findings supported increased genotoxicity in the combination group, aligning with elevated apoptotic responses at both gene and protein levels. In conclusion, the combination of Sorafenib and Favipiravir exhibited notable synergistic, cytotoxic, and anticancer effects on HepG2 cells in vitro. These findings suggest that this drug combination may serve as a promising therapeutic strategy for hepatocellular carcinoma and provides a foundation for further preclinical and clinical studies.