Investigating the cause of reduction of lactate production in Panc-1 cell lines when subjected to digitoxin

Abstract: Pancreatic ductal adenocarcinoma (PDAC) yet remains one of the top 10 most prevalent and top five most common cause of cancer-related deaths solely in the United States. Its metastatic deviation from most cancer types rends it one of the most subtle yet perturbating disease. Hope lies within the traces of reproducibly mutated genes which are said to be found in nearly 95% of all PDAC. This study aimed to examine how digitoxin affects glycolysis and gene expression in Panc-1 cell lines. Panc-1 cells were cultured and treated with various concentrations of digitoxin for 24h-48h depending on the analysis and cell viability was then examined via the MTS assay. Lactate assay was conducted in order to investigate the degree to which panc-1 production of lactate was affected by digitoxin. Specific genes of interest (LDHA, C-Myc, PFKM, PDP1, SLC2A and PMM1) that take part in the glycolytic segment of the panc-1 metabolism were analyzed with qPCR in order to study their expression. Cell cycle assay was conducted in order to examine whether digitoxin also affected the panc-1 cell cycle and to what degree. Results from the MTS assay indicated that cell viability was indeed affected with increasing concentrations of digitoxin whereas; the lactate assay indicated that panc-1 cells were sensitive to 25nM digitoxin/higher due to how the decrease in lactate production became evident at that point. The gene expression of PDP1 was significantly increased with high concentrations of digitoxin compared to other genes. The cell cycle assay indicated that most treatment groups did not make it to the G2 and M phase due to apoptosis. All in all, digitoxin showed to have apoptotic effects on panc-1 cells line although this effect was not evident in certain assays.