Abstract: OBJECTIVE An ionic gelation combined with chemical crosslinking method was developed to prepare chitosan nanoparticles, followed by conjugation with folate (FA) and polyethylene glycol (PEG). METHODS The structures were verified by infrared spectroscopy. The morphology, diameter and Zeta electric potential of the nanoparticles were assayed by environmental scanning electron microscope and scattering particle analyzer. The specificity of the FA+PEG-NPs targeting cancer cells was demonstrated by human adenocarcinoma Hela cells. RESULTS The chitosan NPs presented a narrow size distribution with an average diameter about 200 nm regardless of the type of functional group. Laser confocal scanning imaging proved that both FA+PEG-NPs and FA-NPs could greatly enhance uptake by Hela cells. However, the PEG-NPs showed contrary results. CONCLUSION FA+PEG-NPs can be applied as a new vehicle to actively deliver anticancer drugs to tumor cells.