The purpose of the current study was to evaluate the effect of exogenous addition of vitamin E on the cellular uptake and transport of selenium (Se) across Caco-2 cell monolayer from wheat and pearl millet. Cellular bioavailability of Se was also determined by evaluating the activity of Se dependent enzymes such as glutathione peroxidase (GPx) and thioredoxin reductase (TR). Transport and uptake of Se was by passive diffusion (transcellular transport), which was time dependent. Uptake of selenomethionine (SeMet) and selenocysteine (SeCys₂) by the Caco-2 cells incubated with wheat and pearl millet was significantly enhanced (p < 0.001) in the presence of vitamin E, which, however, was significantly decreased by heat processing (p < 0.001). The mRNA expression of the genes encoding the two Se dependant enzymes, GPx and TR was significantly upregulated in wheat and pearl millet respectively, upon the addition of vitamin E. The extent of upregulation ranged from 80 to 100%.