Correlation Between EGFR And AJAP1 In Glioma And Effects Of EGFR On Cytoskeleton And Invasion And Migration Of Human Glioma U87 Cells

Objective:Through the correlation between EGFR and AJAP1 in glioma and the effect of EGFRV? on U87-MG cytoskeleton and invasion and migration,the new mechanism of EGFR in glioma development and its impact on prognosis were explored.Methods:Immunohistochemistry and immunofluorescence were used to detect the expression of AJAP1 and EGFR in 65 glioma specimens and 10 normal brain tissues.Cellular immunofluorescence assay was used to detect the expression of AJAP1 in U87 and EGFRV? transfected U87 cell line(U87-VIII).Sixty-five patients with glioma were followed up by telephone.The postoperative radiochemotherapy,KPS score,recurrence and death were recorded in detail.Survival curve analysis was performed after follow-up data.Phalloidin was used to detect the changes of U87-VIII cytoskeleton;Western Blot was used to detect the changes of EGFR and proliferation and invasion in U87-VIII;scratch test was used to evaluate the migration ability of U87-VIII;Transwell assay was used to detect the invasive ability of U87-VIII cells.change.Results:Immunohistochemistry and immunofluorescence results showed that the expression of AJAP1 in normal brain tissue was higher than that in low-grade glioma.With the increase of glioma grade,the expression of AJAP1 were decreased,while the expression of EGFR was the opposite.The higher grade the gliomas are,the higher the expression of EGFR;Cellular immunofluorescence showed that AJAP1 was low in U87 and U87-VIII,and lower in U87-VIII,p<0.05,the difference was statistically significant.Statistical analysis showed that the positive rate of AJAP in the high-grade group was significantly lower than that in the low-grade group(30.77% vs 57.69%,p<0.05),and the difference was statistically significant;the expression of EGFR was the opposite(79.49% vs 46.15%,p<0.05);The correlation analysis of AJAP1 and EGFR expression showed a moderate negative correlation(p<0.001,r=-0.535),the difference was statistically significant;The positive expression analysis of AJAP1 and EGFR in high and low grade glioma showed that the positive expression level of AJAP1 in the low-grade group was higher than that in the high-grade group,p<0.05,the difference between the groups was statistically significant,and the expression of EGFR was the opposite.Clinicopathological analysis showed that gender,tumor location,age,and whether the tumor was primary or secondary with AJAP1,EGFR expression was not significantly correlated,p>0.05,the difference was not statistically significant;survival analysis suggested PFS and OS do not have dramatically difference wheather AJAP1 and EGFR positive or not,however,AJAP1 positive survival rate was higher than the negative ones,and EGFR positive survival rate was lower than the negative ones.The cytoskeletal staining of the phalloidin showed that the cytoskeletal structure of the U87-VIII group was changed compared with the U87 group,the filamentous pseudopods of the cells increased,and the slab pseudopods decreased.Western Blot showed that the EGFR,Ki-67,and MMP-9 in U87-VIII were significantly higher than that of U87 and U87-Vector;Transwell test showed that the invasive ability of U87-VIII cells were significantly enhanced;the scratch test showed that the migration ability of U87-VIII cells was enhanced.Conclusion:AJAP1 is negatively correlated with EGFR in gliomas.Low expression of AJAP1 and high expression of EGFR indicate poor prognosis;EGFR can alter the cytoskeleton and enhance the migration and invasion of glioma cells;AJAP1 mediates a novel pathogenesis of EGFR in the development of glioma;...