The Inhibitive Effect Of Rat Hyperplasia Suppressor Gene Onmalignant Glioma Through Ras, Erk1/2 Signal Molecules

Objective To explor mechanism of Ras, Erk1/2 signal molecules in the inhibitive effect of rat hyperplasia suppressor gene(r HSG) on malignant glioma. Methods C6 cells were cultured in vitro and grouped into three groups: A, blank control group(PBS); B, the adenovirus vector control group(Adv-GFP); C, the r HSG over-express group(Adv-r HSG-GFP). Western blot was used to determine the expression of r HSG protein, flow cytometer was used to analyse the effect of r HSG on cell cycle; Western blot was used to determine changes in the expression of p-Erk1/2 protein which could be changed by IGF-1 stimulation in different concentrations and duration. The m RNA expressions of H-ras, N-ras, K-ras, Erk1/2 were identified by QRT-PCR and the protein expressions of H-ras, N-ras, K-ras, Erk1/2, p-Erk1/2, p-Rb, Rb by Western blot after stimulation by IGF-1 in the proper condition. Results After 24 h infected by adenovirus, GFP protein were measured in more than 90% C6 cells.The highest expression of r HSG protein were showed in Group C and the cells were arrested in G0/G1 phase, compare with other two groups there were significantly different(P<0.01). After the 15 minutes’ stimulation of IGF-1 by 10 ng/m L(the proper condition), the highest expression of p-Erk1/2 protein were measured(P< 0.01). After stimulated by IGF-1 in proper condition, there were no difference in the three groups for expressions of m RNA and protein of H-ras, N-ras, K-ras, Erk1/2( P>0.05); expressions of p-Erk1/2, p-Rb protein were the lowest in group C(P< 0.01). For the expression of Rb protein, there were no differences in three groups(P>0.05). Conclusions Our results suggested that the inhibitive effect of r HSG on malignant glioma could be through inhibiting Ras-Raf-MEK-ERK signal pathway.