The Relationship Between WNT/β-catenin Expressioninhibition By End-binding Protein 1 And Autophagy In Cervical Cancer Siha Cells

Objective:Cervical cancer(CC)is one of the most common malignant tumors in women all over the world,and it shows a younger trend in my country.The down-regulation of CC autophagy is closely related to human papilloma virus(HPV)infection in cervical cells and the occurrence and development of cervical cancer.The complete autophagy pathway usually requires four processes:phagocytes,autophagosomes,autolysosomes and degradation.Autophagosome trafficking is a necessary step to achieve vesicle fusion,which enables the degradation process.End-binding protein 1(EB1)plays a key role in connecting microtubules and vesicles.EB1 is a protein that binds to the ends of microtubules,regulates the composition and structural stability of the positive ends of microtubules,and recruits various required proteins.However,it is unclear what role EB1 plays in the transport of autophagosomes to lysosomal sites.Moreover,CC autophagy can lead to the activation and/or inactivation of multiple intracellular signaling pathways,which further leads to disease progression and affects therapeutic effects.Among them,the Wnt/β-catenin signaling pathway plays an important role in the regulation of self-renewal and differentiation of CC cells.This topic mainly studies the interaction between EB1 and cervical cancer cell autophagy and Wnt/β-catenin signaling pathway,and explores that its role may affect cell apoptosis,which is the basis for future research on new cervical cancer early diagnosis and treatment.Target opens up new horizons.Methods:1.The CC cell line Siha cells were cultured in vitro,and the Siha cells were treated with Hank’s balanced salt solution(HBSS)buffer and hydroxychloroquine(HCQ)to induce autophagy in Siha cells and inhibit autophagy in Siha cells.2.Western blot was used to detect the expression of Caspase-3,Bcl-XL,beta-catenin,Beclin-1,P62,Bcl-2 in Siha cells without relevant treatment,HBSS-treated Siha cells and HCQ-treated Siha cells condition.3.Knockdown of EB1 expression by siRNA in Siha cells.4.Western blot was used to detect the expression of LC3-Ⅱ,P62 andβ-catenin proteins in Siha cells with knockdown of EB1 without relevant treatment and after treatment with HBSS and HCQ.Result:1.In this experiment,Siha cells were cultured in vitro,and HBSS was used to induce Siha cell autophagy;HCQ was used to inhibit Siha cell autophagy.The results showed that compared with untreated Siha cells,the expression levels of Beclin-1 and P62 proteins increased significantly after HBSS treatment,and the difference was statistically significant(P<0.01).After Siha cells were treated with HBSS and HCQ,compared with untreated Siha cells,the expression level of β-catenin protein was significantly decreased,and the difference was statistically significant(P<0.01).After Siha cells were treated with HCQ,compared with untreated Siha cells,the expression levels of caspase-3 and P53 proteins were significantly increased,and the expression levels of Bcl-XL and Bcl-2 proteins were significantly decreased,and the differences were statistically significant(P<0.05).2.When the expression of EB1 in Siha cells was inhibited by siRNA,the expression levels of P62 and LC-3Ⅱ proteins were significantly increased compared with those in non-knockdown Siha cells.When combined with HBSS treatment,the changes in the expression of LC3-Ⅱ and p62 were the same as those in Siha cells that only inhibited EB1,resulting in an increase in the expression of LC3-Ⅱ and p62.When the Siha cells with knockdown of EB1 were treated with HCQ in combination with the Siha cells without knockdown of EB1 treated with HCQ,the expression level of LC-3Ⅱ protein was further enhanced,and the expression of p62 protein did not change significantly.3.After knockdown of EB1 in Siha cells,the expression level ofβ-catenin protein increased in NC group,HBSS and HCQ without related treatment.Conclusion:Autophagy of cervical cancer Siha cells can affect the transduction of Wnt/β-catenin signaling pathway,EB1 may be involved in the formation of autophagosome,and influence the combination of autophagosome and lysosome to form autophagosome-lysosome,and inhibitβ-catenin expression.The Wnt/β-catenin signaling pathway and autophagy regulation in cervical cancer may be global,not just in the case of a certain external environment.