Rap1GAP Protein Effects On The Proliferation,Migration,Invasion In Glioma Cells

Objective: This study was undertaken to define the expression and function of Rap1GAP in glioma cells.Methods: Firstly, the expression and function of Rap1GAP in glioma cells were studied. Western Blot and cell immunofluorescence were used to detect the expression of Rap1GAP in glioma cell lines U87, U251, U118. Overexpression of Rap1GAP gene virus was constructed. Virus were used to infected glioma cells, and the carrier group and the control group were established. After infected, Western blot were used to detect the expression of Rap1GAP. CCK-8 and flow cytometry were used to determine the growth of glioma cell lines with overexpression of Rap1GAP. Then we investigated the effect of Rap1GAP on glioma cells invasion and migration, we used Transwell chamber to detecte cells invasion and migration, and we used western blot and RT-PCR to find the expression of MMP2 and MMP9.Then we detected the reason of Rap1GAP downregulated in glioma cells, we used the demethylation agent 5-aza, 50umol/L, 72h, then used PCR detection. Flow cytometry was used to determine the apoptosis and cell cycles.Results:The results showed Rap1GAP was low in glioma cell lines((P<0.05).The flow cytometry showed that compared to the control group, overexpression of Rap1GAP in glioma cells group significantly inhibited the cells, growth, inhibiting GO phase cells((P<0.05), and western blot showed that Rap1GAP also reduced the Rap1,Rap2 and ERK phosphorylation protein levels((P<0.05). It reduced cells’ ability to migrate ((P<0.05), but enhanced invasion and the MMP2 and MMP9 levels ((P<0.05).Finally, in order to understand the reason of Rap1GAP downregulated in glioma cells, we used the demethylation agent 5-aza, 50umol/L, 72h, then used PCR detection. Results showed the normal group ratio of target gene expression levels were significantly lower than dosing group (P<0.05).Conclusion:1、Rap 1 GAP was low expressed in glioma cell lines,U251,U87, U118.2、Overexpression of Rap 1 GAP inhibited cell proliferation,apoptosis and migration.3、Rap 1 GAP promoted glioma cells invasion.