Pancreatic cancer is the ninth leading cause of cancer death in women and the eight in men. There are different ways in which digitoxin can induce apoptosis in cancer cell lines in vitro, as they are said to have antiproliferative effects on cancer cells. Hyperglycemia through oxidative stress can increase the ROS levels of the cancer cells thereby promoting proliferation of pancreatic cancer cell. T-cells are important when considering the immune response of neoplasm and are considered dysfunctional when they express the programmed cell death protein 1, thus the use of keytruda could prove helpful. The aim of this study is to show the effects of digitoxin, pembrolizumab and combination of both on cell proliferation of ASPC-1 cancer cell line using MTS assay, also to show the effects on gene expression using keytruda, digitoxin and [Digitoxin+Keytruda] by performing qPCR. The report also aims to investigate the effects on the viability of the cells by combining digitoxin and pembrolizumab on ASPC-1 cancer cell line as well as to observe the ROS levels using Amplex red assay. Cells were harvested, counted, seeded and treated. After which MTS assay and Amplex red assay reading were performed. RT-qPCR was performed to observe expressions of PD1, PDL1 and PDL2 genes. It was observed that cell proliferation decreased with the treatments, as well as decrease in ROS levels. Lastly, gene expression for PD1 was affected when cells were treated with [1nM + 50mg/l] treatment by lowering the expression of PD1 gene.