Cancer is one of the most common causes of death worldwide. Inability in controlling anticancer drug delivery to tumor, systemic side effect on healthy tissue, instability of anticancer drugs, low concentration of drug around the tumor, and low efficiency of drug effect on the solid tumor may cause high mortality rate in cancer patients. Drug delivery systems based on nanofibers could perform an approach to treat cancer with the least side effects. The aim of this study is to improve a nanofiber drug delivery system based on polycaprolactone/chitosan blended electrospun nanofibers containing 5-fluorouracil. Nanofiber characteristics and drug release behavior of various nanofibers were investigated. The polymeric proportion had significant effect on release behavior, mechanical properties, and nanofiber morphology. Loading efficiency of these nanofibrous drug delivery systems was good, and its rate increased by increasing chitosan proportion. Also, increasing the chitosan in nanofibers led to increasing drug release period. The release mechanism of all nanofibers was Fickian diffusion according to Korsmeyer-Peppas model. This nanofibrous drug delivery system had a great potential to perform an anticancer drug delivery system for colorectal cancer. Further studies need to be conducted to improve the release period of 5-fluorouracil from these nanofibers.