Högskolan i Skövde

Hepatocellular carcinoma (HCC) is the main disease affecting the liver and widely extended over the population, with a prevalence that has increased over the years. Several studies have shown the effect of digitoxin (a cardiac glycoside) and pembrolizumab (an immunotherapy antibody) in reducing the viability of cancer cells by promoting apoptosis and increasing the action of T-cells against them, respectively. In this project, a study was conducted in order to assess the effect of these two drugs on the HepG2 cancer cell line after 24, 48, and 72h of treatment. Different concentrations of the drugs were tested to identify the existence of any significant difference in the cell’s viability with 1, 5, 10, 25 and 100 nM of digitoxin and 0.5, 2, 10 and 50 mg/L of pembrolizumab and their combination. An MTS assay was performed as well as an Amplex-Red assay in order to measure the cell viability and the ROS (reactive oxygen species) levels. Finally, the expression of PD-1, PD-L1, and PD-L2, important genes for the tumor progression, was measured performing a RT-q-PCR. The results showed that certain concentrations of the drugs promoted a decrease in the cell viability, indeed, showing synergetic effects when combined. Moreover, the expression of the genes was confirmed, obtaining a significant variability depending on the treatment. Finally, ROS levels decreased after the treatment at specific concentrations.