GPX2 Suppression Of ROS Levels Regulates The Cervical Cancer Metastatic Via Activation Of The β-catenin/WNT Pathway

Objective:To investigate the expression of Glutathione peroxidase 2(GPX2)in cervical cancer tissues and cells and the correlation between GPX2 and the clinicopathological features of cervical cancer.Using molecular biology techniques to explore the relationship of GPX2 and Reactive oxygen species(ROS)and whether via activating Epithelial to mesenchymal transition(EMT)and β-catenin/WNT pathway that affect tumor cell proliferation,metastasis and other biological characteristics,reveal the pathogenesis of cervical cancer,and provide new targets for clinical treatmentMethods:Transient transfection,real-time fluorescence quantitative PCR and Western blot were used to construct overexpressed and low-expressed GPX2 cervical cancer cells,respectively.The changes in the biological phenotypes(proliferation,migration and invasion)of ME180 and He La of cervical cancer cells before and after GPX2 transfection were detected by clone formation assay,Transwell and other experimental techniques,then the intracellular ROS level was detected by flow cytometry.A series of m RNA and protein levels were used to determine whether changes in cell biology were related to EMT,and whether GPX2 regulated EMT via the classical β-catenin /WNT signaling pathway.Finally,the levels of GPX2 in clinical cervical tissues were detected by immunohistochemistry and the relationship between GPX2 expression and distal metastasis of cervical cancer was analyzed.Result:1.TCGA database was used to analyze GPX2 m RNA expression levels in cervical squamous cell carcinoma(CESE)and normal cervical tissue samples.The results showed that GPX2 expression was significantly increased in CESE samples,and the difference was statistically significant(P < 0.001).2.The results of the migration and invasion of cervical cancer cells showed that,compared with the Control group,the migration and invasion abilities of the cells in the ME180 low-expression GPX2 group(sh GPX2#5 and sh GPX2#8)were significantly inhibited,and the differences were statistically significant(P < 0.001).Compared with the control group(Pex-6),the He La overexpressed GPX2 group(GPX2-Pex-6)showed significantly enhanced cell migration and invasion ability,and the difference was statistically significant(P < 0.001).3.After cervical cancer cells were transfected with GPX2,the m RNA and protein levels of EMT-related markers E-cadherin(epithelial cell marker)and Vimentin(mesenchymal cell marker)were significantly different compared with the control group.In ME180 cells,the m RNA and protein levels of E-cadherin were significantly higher in sh GPX2#5 and sh GPX2#8 cells than the control group,while the m RNA and protein levels of Vimentin were significantly lower.In He La cells,the m RNA and protein levels of E-cadherin in the GPX2-Pex-6 group were significantly lower than those in the control group,while the m RNA and protein levels of Vimentin were significantly higher.The difference was statistically significant(P < 0.001).4.In ME180 cells,ROS levels of sh GPX2#5 and sh GPX2#8 cells were significantly higher than those of control cells.On the contrary,in He La cells,the ROS level of GPX2-Pex-6 group was significantly lower than that of the control group,and the difference was statistically significant(P < 0.001).5.The m RNA expression levels of TCF-1 and cyclin D1 in the classical β-catenin /WNT signaling pathway were significantly different after cervical cancer cells were transfected with GPX2.In ME180 cells,the m RNA expression levels of TCF-1 and cyclin D1 were decreased in sh GPX2#5 and sh GPX2#8 cells compared with the control group.In He La cells,the m RNA expression levels of TCF-1 and cyclin D1 in GPX2-Pex-6 group were increased.The difference was statistically significant(P < 0.001).6.Immunohistochemical analysis of cervical tissues with different pathological grades showed that,compared with normal cervical tissues,GPX2 expression in cervical tissues showed a significant positive correlation with pathological grades,and the difference was statistically significant(P < 0.001).Conclusion1.GPX2 can promote the migration and invasion of cervical cancer cells,suggesting that GPX2 can accelerate the metastasis of cancer cells during the development of cervical cancer and provide new therapeutic targets for the clinical treatment of cervical cancer.2.GPX2 can consume the ROS level in cervical cancer cells,reduce intracellular oxidative stress,promote tumor growth and is related to the EMT/ β-catenin/WNT pathway,providing a molecular mechanism for the clinical treatment of cervical cancer3.The GPX2 expression level in the tissues of patients with cervical cancer is related to the pathological stage of cervical cancer and lymph node metastasis,suggesting that the GPX2 expression level in the tissues of cervical cancer can reflect the progress of cervical cancer and the prognosis evaluation.