Abstract: Objective To explore the relationship of hyperthermia with heat shock protein 70(HSP70) and induced nitric oxide synthase(iNOS) expression and to clarify hyperthermia reducing glioma invasiveness and its possible mechanism. Methods After heat treatment on glioma cells C6, we determined HSP70 antibody titer and the expression level of iNOS by ELISA and Western blot; Nitric oxide(NO) concentration in glioma cells were determined by Griess; We used crystal violet staining method to determine the glioma invasion. Results HSP70 antibody titer in C6 cells was increased and reached the maximum at 30 min(86.52ng/L) after hyperthermia. The expression level of iNOS in C6 cells was also increased significantly and reached the peak at 60min(IDV=228.31) after hyperthermia, then gradually decreased. NO concentration of glioma cells reached the peak at 120min(OD=168.42) after hyperthermia. The higher NO concentration was, the lower glioma invasion was(n=8.21×10⁶). Conclusion Hyperthermia increases HSP70 expression in glioma cells. HSP70 probably improves the expression level of iNOS by a mechanism, thereby increases the concentration of NO in glioma cells and decreases the glioma invasion.