Electrospinning is a simple process for the production of fibers with diameters in the range from submicron to micron. Herein we aim to explore the influence of fibrous diameter on the drug delivery. The feasible methods by making choice of solvents and changing flow rate were used to prepare 5-fluorouracil-loaded polylactide (PLA) fibers with a large diameter gap. The drug release behavior in vitro was investigated and analyzed in phosphate buffer solution. The drug distribution and fiber diameter both affected the initial burst release. The results showed that all the asspun fibers could not avoid of burst release. The coarse fibers exhibited slight burst release as compared to fine fibers. During the second stage, the fine fibers released faster than that of the coarse fibers. For the whole release stage, the large-diameter fibers seemed to be beneficial for drug release in the long term and smoothly. The MTT results showed that the cytotoxicity of drugs was maintained.