Högskolan i Skövde

Pancreatic Ductal Adenocarcinomas (PDAC) is the fourth deadliest cancer in western countries, 95% of diagnosed patients have 1 to 5 months of survival chance, and only 5% survive 5 years. The high morbidity and mortality can be due to late diagnosis which leads to a poorly prognosis, most of PDAC patients are diagnosed metastatic stages, with median age of 60-70 years old. Risk factors that are associated with tumorigenesis of PDAC: environmental factor, genetic mutation, elevated Reactive Oxygen Species (ROS), suppression of Programmed death receptor (PD-1) by PD-ligands (PD-L1 and PD-L2). The aim of the study was to investigate if proliferation of CFPAC-1 cells from PDAC, grown in vitro could be inhibited by Digitoxin, to investigate the role of Digitoxin on ROS production in CFPAC-1, and also investigate if combination of Digitoxin and Pembrolizumab leads to synergistic effect. The expression of PD-1 and PD-L1/L2 were investigated by Quantitative Polymerase Chain Reaction (qPCR). The MTS-assay was used to determine the cell viability, the Amplex-Red-assay was used to investigate the ROS level in CFPAC-1. Found was that the cell proliferation was significantly inhibited by Digitoxin at 10-100nM concentration, and also by the combination of Digitoxin with Pembrolizumab. The ROS production was upregulated by 100nM Digitoxin and down-regulated by 25nM Digitoxin and combination of Digitoxin with Pembrolizumab. The expression of PD-1, PD-L1/L2 were upregulated by 10-and-50mg/L Pembrolizumab, and by the combination of Pembrolizumab and 100nM Digitoxin. These findings suggest that Digitoxin has an anti-proliferation effect and can be used in combination with Pembrolizumab as anti-cancer therapy for PDAC.