Effect And Significance Of IS201 On FAK-ras/MAPK Signaling Pathway Of GL15 Glioma Cell

Objective As we know,Integrinαvβ3 can regulates FAK-ras/MAPK singnal pathway that play a main role on biological behavior in GL15 glioma cell. IS201,as a potent antagonist of integrinαvβ3, can specifically recognize the RGD sequence and bind, thus to influence the integrin-mediated signal transduction. The main point of our experiement is to observe the influence of protein belonging to FAK-ras/MAPK singnal pathway expression and phosphorylation after IS201 binding integrinαvβ3.To a certain extent, IS201 can reduce the invasion of tumor cells thus being great help to cancer treatment.Methods (1) GL15 glioma cells cultured in DMEM with different concentrations (0mg / L, 20mg / L and 40mg / L) of the IS201. Samples and lysates were collected after 16 hours incubation. (2) The sample line SDS-PAGE, and transferred to PVDF membrane, blocking solution at room temperature closed, then added to the primary antibody (1:1000 dilution of anti-FAK polyclonal antibody and pho-FAK antibody; MMP-2 antibody 1:1000 dilution; ERK1 / 2 phosphorylation antibody 1:1000 dilution) and horseradish peroxidase labeled goat anti-rabbit secondary antibody at room temperature for 1 hour developing, actin as a protein sample volume calibration. (3) Established Transwell model plate, cultured 12h, the cells through Matrigel membrane porous plastic and PET and to the next surface. Cell fixation, stained and calculated of the number of invasive cells .Results (1) Different concentrations of IS201 could inhibit FAK expression (* P <0.05) and FAK phosphorylation (* P <0.05), and with the IS201 concentration increased, its expression and phosphorylation of FAK was more inhibited. (2) By adding IS201 in all experimental groups, ERK1 / 2 expression was significantly decreased (* P <0.05), with the IS201 concentration is increased, its inhibitory effect on ERK1 / 2 showed a increasing trend. (3) Different concentrations of IS201 on the MMP-2 were significantly reduced (* P <0.05), with the IS201 concentration increased, its inhibitory effect on MMP-2 showed a increasing trend. (4) Different concentrations IS201 have influence on the GL15 cell invasion, with the IS201 concentration increased, its inhibitory effection increasing .Conclusion Integrinαvβ3 antagonists IS201 can inhibits FAK expression , FAK phosphorylation , MMP-2 expression, ERK1 / 2 expression related with FAK-ras/MAPK signal pathway in GL15 glioma cells. It can also inhibits the invasion of GL15 cells significantly . with the IS201 concentration increased,its inhibitory effction showed a increased trend.