| Objectives To study the expression levels of ERP29,AKT,pAKT,mTOR,pmTOR,4EBP1 and p4EBP1 in ovarian tissues of patients with epithelial ovarian cancer,and to explore the relationships between their expression levels and clinicopathological characteristics.To analyze the effect of ERP29 on the proliferation,invasion and apoptosis of ovarian cancer cells,and its possible significance in the development of epithelial ovarian cancer.Methods 1 37 cases of ovarian tissues from epithelial ovarian cancer patients who were surgically treated and received pathological confirmation in the Obstetrics and Gynecology Department of Tangshan Gongren Hospital from November 2018 to September 2019 were selected,meanwhile 32 normal ovarian tissues and 32 benign ovarian tumor tissues were collected as controls.Western blot was used to detect the expression levels of ERP29,AKT,pAKT,mTOR,pmTOR,4EBP1 and p4EBP1 in ovarian tissues of each group,and to analyze the relationships between the expression levels of ERP29,AKT,pAKT,mTOR,pmTOR,4EBP1,p4EBP1 and the clinicopathological characteristics of patients,and to analyze the correlations between ERP29 and AKT,pAKT,mTOR,pmTOR,4EBP1,p4EBP1.2 Human ovarian cancer SKOV3 cells were cultured in vitro,and the ERP29 gene was silenced by cell transfection technique.After the cells' model was successfully constructed,it was used as the experimental group(siRNA-ERP29 group),SKOV3 cells transfected with blank vector siRNA-NC as the negative control group(siRNA-NC group),and untreated SKOV3 cells as the blank control group(blank group).The expression levels of AKT,mTOR and 4EBP1 mRNA in each group of cells were detected by RT-qPCR,and the expression levels of ERP29,AKT,pAKT,mTOR,pmTOR,4EBP1 and p4EBP1 in each group of cells were detected by Western blot,the CCK-8 test was used to detect the proliferation ability of each group of cells,the scratch test was used to detect the invasion ability of each group of cells,and flow cytometry was used to detect apoptosis in each group.3 SPSS 17.0 software was used to analyzed the data obtained by the experiments.Results 1 The expression of ERP29 in the epithelial ovarian cancer group was significantly lower than that in the benign tumor group and normal ovarian group,and the difference between the three was statistically significant(F=3052.247,P<0.05);the expression levels of AKT,pAKT,mTOR,pmTOR,p4EBP1 in the epithelial ovarian cancer group were significantly higher than that in the benign tumor group and normal ovarian group,and the differences between the three were statistically significant(F=542.727,F=902.637,F=169.081,F=420.940,F=284.819,P<0.05);the expression level of 4EBP1 in epithelial ovarian cancer group,benign tumor group and normal ovarian group had no statistically significant difference(F=1.299,P>0.05).In epithelial ovarian cancer tissues,the expression level of ERP29 was related to FIGO stage and lymphatic metastasis(P<0.05),and had nothing to do with the age,pathological grade,histological type and ascites(P>0.05);the expression levels of AKT,pAKT,mTOR,pmTOR,and p4EBP1 were related to FIGO stage,pathological grade,and lymphatic metastasis(P<0.05),and had nothing to do with the age,histological type,and ascites(P>0.05).In normal ovarian tissues,benign tumor tissues and epithelial ovarian cancer tissues,ERP29 was not significantly correlated with the expression levels of AKT,mTOR,and 4EBP1(P>0.05),but negatively correlated with the expression levels of pAKT,pmTOR,and p4EBP1(P<0.05).2 Cell experiment results showed that the expression levels of AKT,mTOR,4EBP1 mRNA in siRNA-ERP29 group were significantly higher than that in blank group and siRNA-NC group(F=994.687,F=3176.362,F=7.533,P<0.05),there were no statistically significant differences in the expression of the three in the blank group and the siRNA-NC group(P>0.05).The expression levels of AKT,mTOR,4EBP1 in siRNA-ERP29 group,blank group and siRNA-NC group were not statistically different(F=2.261,F=0.864,F=2.337,P>0.05);the expression levels of pAKT,pmTOR,p4EBP1 in siRNA-ERP29 group were significantly higher than that in blank group and siRNA-NC group(F=1256.980,F=1032.555,F=50.572,P<0.05),there were no statistically significant differences in the expression levels of the three in the blank group and the siRNA-NC group(P>0.05).48 hours after the completion of cell transfection,the cells in the siRNA-ERP29 group showed stronger proliferation ability than the blank group and the siRNA-NC group(F=35.485,P<0.05).Compared with the blank group and the siRNA-NC group,the siRNA-ERP29 group had stronger invasive ability(F=52.779,P<0.05).Compared with the blank group and the siRNA-NC group,the apoptosis rate of the siRNA-ERP29 group decreased,but there was no statistically significant difference(P>0.05).There were no statistically significant differences in cells' proliferation,invasion and apoptosis ability between blank group and siRNA-NC group(P>0.05).Conclusions 1 With the evolution of the disease,the expression of ERP29 is reduced in epithelial ovarian cancer tissues,while AKT,pAKT,mTOR,pmTOR,p4EBP1 are increased in epithelial ovarian cancer tissues,and the expression of 4EBP1 in epithelial ovarian cancer tissues is not obvious changed.2 ERP29 is negatively correlated with the expression of pAKT,pmTOR,p4EBP1,but has no significant correlations with AKT,mTOR,4EBP1.3 Silencing ERP29 can increase the expression levels of pAKT,pmTOR and p4EBP1 in SKOV3 cells and enhance the proliferation and invasion ability of SKOV3 cells,but it has no significant effect on the expression of AKT,mTOR,4EBP1 and the apoptosis ability of SKOV3 cells.4 ERP29 plays a role in the occurrence and development of ovarian cancer as a tumor suppressor,and its mechanism is related to the inhibition of mTOR pathway activation.Figure 10;Table 22;Reference 138... |
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