Combinatory effects of sorafenib and berberine treatment on hepatocellular carcinoma cell line, hep3b

GLOBOCAN 2018 data has displayed liver cancer to be the fourth most deadly type of cancer worldwide. HCC carcinoma being the most common type of liver cancer accounts for 80 percent of the deaths. Berberine (BBR) is an alkaloid derivative and used as a herbal medicine. The BBR effective drug in liver diseases and some cancers. Sorafenib (SOR) is a multi-tyrosine kinase inhibitor and used for different cancer treatments, especially in HCC. However, SOR treatment could increase the levels of HIF-1α, which leads to increased chemoresistance in cancer therapy. In this study, SOR was combined with BBR in order to enhance the efficacy of SOR therapy. The results of BBR and co-treatment with SOR on the Hep3B cell line have been observed by using MTS assay. The combination effects of BBR and SOR were calculated using CompuSyn and BBR and SOR were also selected for the co-treatment. After 25 μM BBR treatment, the viability of Hep3B cells decreased to 75 percent while the cell viability was decreased to 25 percent in the combination of 25 μM BBR and 20 μM SOR. The caspase cascade protein markers and Annexin V-FITC / PI staining have also been used to identify stages of cell death. The BBR treatment induced early apoptosis (21 percent) in the Hep3B cells and the combination treatment triggered late apoptosis (51.24 percent). The HIF-1α, LDHA protein expression levels were reduced to 78.66 and 76.41 percent respectively after BBR treatment and in co-treatment the HIF-1α, LDHA protein expression level was 80 and 90.27 percent of the control. AMPK activation also increased to 518 percent in the BBR treatment. The inner mitochondrial membrane potential was quantified by flow cytometry and observed by confocal microscopy. Moreover, all OXPHOS complexes showed a significant decrease following BBR application under hypoxic conditions. In conclusion, BBR downregulates the protein expression level of HIF-1α, reduces the effect of Warburg, and induces apoptosis. Due to these characteristics, BBR may be used as a supporting therapy particularly for SOR-resistant cancers.