Choline is an organic cation that plays a critical role in the structure and function of biological membranes. Intracellular choline accumulation through choline transporters is the rate-limiting step in phospholipid metabolism, and it is a prerequisite for cell proliferation. Choline PET/CT has been used to visualize various cancers, and high levels of choline accumulation have been observed in tumors. However, the uptake system for choline and the functional expression of choline transporters in pancreatic cancer are not completely understood. In this study, we examined the functional characterization of choline transporters in pancreatic cancer cells. Furthermore, we searched for compounds that inhibit choline uptake as well as cell proliferation in a plant-derived natural organic compound library.