The Effect Of Irisin On The Malignant Biological Behavior Of Epithelial 0varian Cancer And Its Mechanism

ObjectiveTo study the expression of irisin/FNDC5 in epithelial ovarian cancer tissues,and to investigate the effect of irisin on the malignant biological behavior of A2780 and SKOV3 cells and its mechanism.MethodsImmunohistochemical experiments were used to detect the expression of irisin/FNDC5 in 40 epithelial ovarian cancer tissues and 10 normal ovarian tissues.The tissues were collected from patients hospitalized for surgical treatment in Qingdao Municipal Hospital Affiliated to Qingdao University from January 2018 to January2020.Epithelial ovarian cancer A2780 and SKOV3 cells were treated with different concentrations of irisin.CCK-8 assay were used to detect cell proliferation,scratch assay to detect cell migration,invasion assay to detect cell invasion,Western blot assay to detect AKT,p-AKT,E-Expression of cadherin,N-cadherin,Vimentin and Snail proteins.PI3 K agonist(740Y-P)combined with irisin was used to treat epithelial ovarian cancer A2780 and SKOV3 cells,and the cell function experiments were repeated to further verify the specific mechanism of the effect of irisin on the malignant biological behavior of epithelial ovarian cancer cells.Results1.The positive rate of irisin/FNDC5 in epithelial ovarian cancer tissues was87.50%(35/40),significantly higher than that in the control group 30.00%(3/10)(P<0.001).Its expression level was correlated with FIGO stage(P=0.022)and lymph node metastasis(P=0.012),but not with age(P=0.356),pathological type(P=0.338)and histological grade(P=0.142).2.The results of CCK8 proliferation experiment,scratch experiment and Transwell invasion experiment showed that irisin inhibited the proliferation,migration and invasion of A2780 cells at both physiological and pharmacological concentrations(P<0.05),but only inhibited SKOV3 cells at pharmacological concentrations(P<0.05).3.Western blot results showed that 48 hours after treating epithelial ovarian cancer cells with different concentrations of irisin,The expression of epithelial marker Ecadherin protein was higher than that of the control group,while the expression of interstitial marker Vimentin,N-cadherin and E-cadherin protein transcription inhibitor Snail protein was lower than that of the control group,and both physiological and pharmacological concentrations of irisin reduced the amount of phosphorylated AKT in A2780 cells without changing the amount of total AKT(P<0.05).But irisin only have the same effect on SKOV3 cells at pharmacological concentrations(P<0.05).4.After the addition of PI3 K agonist(740Y-P),Western blot results showed that compared with irisin alone,740Y-P upregulated the amount of phosphorylated AKT,partially reversed the expression of EMT-related proteins,and partially reversed the progression of EMT inhibited by irisin(P<0.05).Scratch test and Transwell invasion test showed that 740Y-P partially reversed the inhibition of irisin on migration and invasion(P<0.05).Conclusion1.The expression level of irisin/FNDC5 in epithelial ovarian cancer tissues were higher than that in normal ovarian tissues,and its expression level was related to the FIGO stage and lymph node metastasis of ovarian cancer;2.Irisin can inhibit the migration and invasion of A2780 and SKOV3 cells in a concentration-dependent manner;3.Irisin can induce the EMT process of A2780 and SKOV3 cells.4.The PI3 K agonist 740Y-P partially restored the inhibitory effects of irisin on ovarian cancer cell invasion,migration and EMT.