Rho/ROCK Inhibitor, Fasudil, Suppresses Glioma Cell Line SHG44 Progression In Vitro

Objected:To explore the anti-tumor effects (proliferation, migration and apoptosis) of Rho/ROCK inhibitor, fasudil, including the possible mechanisms involved in the suppression of the human glioma cell line SHG44 progression.Method:After SHG44 were treated with various concentrations of fasudil(10,20,40,80, 160μmol/L), the effects of ROCK inhibitor, fasudil, on proliferation, migration,and apoptosis of cultured tumor cells SHG44 were examined. Effect of Fasudil on the morphological changes of SHG44 cells was observed by phase microscope. MTT was performed to examine the effect of various concentrations of fasudil on the proliferation of glioma cells after treated in 16,24,48h. The wound healing assay was used to assess the effects of fasudil on the migration capacity of SHG44 cells. The sites of the wound line were photographed immediately after scratching and 24h later. The migration areas was measured by Image J. Annexin V/PI staining assay was used for investigating the ability of the fasudil to induce SHG44 cells apoptosis after 24h. Statistic package of SPSS 13.0 was used for the data analysis and significant differences between means were evaluated by ANOVA analysis.Result:1. The proliferation of SHG44 cells was notably attenuated,observed by MTT colorimetric survival assay. IC50 of 16,24,48 hour were 107.12μM/L,79.07μM/L,66.88μM/L respectively (P<0.05). Statistical analysis revealed treatment with fasudil inhibited growth of SHG44 cells in a time and dose-dependent manner; 2.The wound healing assay indicated that Fasudil can inhibit cell migration of human astrocytic glioma cell line SHG-44 in a dose-dependent manner. There was significant difference between higher-concentration groups (40,80,160μmol/L) and lower-concentration groups (10,20μmol/L) or control group (P<0.05).3. Flow cytometry demonstrates that fasudil induced the cell apoptosis of shg44 cells over 24 h in a dose-dependent manner. Compared with the lower-concentration groups (10,20μmol/L) or control group, early apoptosis rate of the cells in higher-concentration groups (40, 80,160μmol/L) increased markedly in a dose-dependent manner. Conclusion:Based on the results, fasudil may suppresses the progression of SHG44 in vitro by inhibiting ROCK. On the other hand,this also could be linked to the induction of apoptosis in tumor cells. Therefore,the Rho/ROCK signaling pathway may prove to be one of promising targets in anti-tumor therapy in future.