Expression Of Robo4 In Human Endometrial Cancer And The Effect On Its Biological Behavior

Objective:To study the relationship between Robo4 and human Endometrial Cancer(EC),to study the effect of Robo4 on the malignant biological behavior of EC cells in vitro,and to explore the mechanism of Robo4 in the Genesis and development of EC.Materials and methods:In Part Ⅰ,100 tissue samples and serum samples(60 EC,20 Atypical Hyperplasia of Endometrium(AHE)and 20 Normal Endometrium(NE)were collected from Tianjin Central hospital of Gynecology and obstetrics.The expression of Robo4 protein in endometrial tissues was detected by Immunohistochemistry,and the expression of Robo4 protein in serum was detected by enzyme-linked immunosorbent assay(ELISA),survival analysis was used to explore the relationship between the expression of Robo4 and the prognosis of EC patients.Vasculogenic Mimicry(VM)in EC tissue was detected by using CD31 and PAS double staining.The relationship between the expression of Robo4 and Vasculogenic Mimicry(VM)in EC tissue and the relationship between VM and pathological features of EC were studied by statistical analysis.In Part Ⅱ,we selected both Ishikawa cells and AN3CA cells to knock down and over-express the Robo4 gene respectively in vitro.We detected the two kinds of cell’s proliferation ability by CCK-8 kit test,migration ability by Cell Scratch test and Invasion ability by Transwell cell invasive test.RESULT:1.There was no significant difference in the expression of Robo4 in NC,AHE and EC patients’s serum(P=0.202).2.The expression of Robo4 in EC and AHE tissues were significantly higher than that in NC tissues(P<0.01).The expression of Robo4 in early and middle stage(Ⅰ-Ⅱ stage),Type Ⅰ and low grade EC tissues was significantly higher than that in late stage(Ⅲ-Ⅳ stage),Type Ⅱ and high grade EC Tissues(P<0.05).3.The Overall Survival time(OS)and Disease-free Survival(DFS)in Robo4 high-expression Group were significantly higher than those in Robo4 low-expression Group(P<0.05).4.The positive rate of VM in Robo4 low-expression Group was higher than that in Robo4 high-expression Group(P<0.05),and the positive rate of VM in high-grade and late-stage(Ⅲ-Ⅳ)of EC tissues was higher than that in low-grade and early-middle-stage(Ⅰ-Ⅱ)of EC tissues(P<0.05).5.The expression of Robo4 in Ishikawa cells was high,while the expression of Robo4 was low in AN3CA cells(P<0.05).After knocking down the expression of Robo4,the proliferation,migration and invasion ability of Ishikawa cells was obviously decreased(P<0.05),there was no obvious change in the proliferation,migration and invasion ability of AN3CA cells after overexpression of Robo4.Conclusion:1.The expression of Robo4 in low grade and type Ⅰ EC tissue was higher than that in high grade and Type Ⅱ EC tissue;2.The expression of Robo4 was negatively correlated with VM in EC tissue;3.Robo4 promotes the malignant biological behavior of Ishikawa cells,and has no effect on the malignant biological behavior of AN3CA cells.