The blending of poly(N-isopropylacrylamide)-co-polystyrene nanofibers with functionalized gold nanoparticles was explored to form the new nanocomposites to facilitate the accumulation of daunorubicin inside leukemia cells and potentially enhance the cure efficiency. The synergistic enhancement effect of the nanocomposites on the drug uptake of daunorubicin in drug resistant leukemia K562 cells has been observed by using electrochemical and confocal fluorescence microscopy studies. The results suggest that these nanocomposites could be utilized to facilitate the drug delivery and realization of the early diagnosis of the respective cancer cells, suggesting the potential valuable application in the relative biomedical and bioanalysis areas.