The present studies reveal that silver nanoparticles (AgNPs) can induce apoptosis and enhance radio-sensitivity on cancer cells. In this paper, we mainly investigated the effect of AgNPs on rat glioma C6 cells upon the combination treatment of hyperthermia treatment (HTT). AgNPs were synthesized by a polyol process and the mean size was 15 nm. The particles showed dose-dependent cytotoxicity on C6 cells from the experimental data. Besides, we found that heating cells could enhance the contents of cell uptake of AgNPs. From the survival curves, AgNPs showed the ability to enhance thermo-sensitivity on C6 cells. Our results revealed that AgNPs could have a potential application in enhancing effect on HTT induced killing of glioma cells.