In this work, 20(S)-Protopanaxadiol (PPD) loaded polycaprolactone (PCL) nanofibers were successfully fabricated by electrospinning technique using tween-80 as a solubilizer. Firstly, smooth and continuous nanofibers were collected by using suitable solvent and appropriate spinning conditions. Secondly, nanofiber mats were characterized by scanning electron microscope (SEM), thermogravimetric (TG) analysis, Fourier transform infrared spectroscopy (FTIR) and mechanical test. Finally, nanofibrous membranes were evaluated using water contact angle, in vitro drug release, biodegradation test, in vitro and vivo antitumor activity and cell apoptosis assay. As a result, scanning electron microscopy observations indicated that diameter of the drug-loaded nanofibers increased with drug concentration increasing. Thermogravimetric (TG) analysis and mechanical test showed nanofibers equipped with great thermal and mechanical properties. Biodegradation test exhibited that the structure of fabricated nanofibers had a certain degree of change after 15 days. In vitro release study showed that PPD from drug-loaded nanofibers could be released in a sustained and prolonged mode. The cytotoxic effect of drug-loaded nanofiber mats examined on the human laryngeal carcinoma cells (Hep-2 cells) demonstrated that the prepared nanofibers had a remarkable antitumor effect. Meanwhile, the drug-loaded fiber mats showed a super antitumor effect in vivo antitumor study. All in all, PCL nanofibers could be a potential carrier of PPD for cancer treatment.