A Preliminary Study On The Targeting Of ZWINT Against Epithelial Ovarian Cancer

Background and purposes:Ovarian cancer is the deadliest primary malignant tumor of the female reproductive system.Because the early symptoms of ovarian cancer are not obvious,about 70% of patients are clinically diagnosed as advanced and lose the chance of radical treatment.If ovarian cancer can be diagnosed at an early stage,the 5-year relative survival rate will exceed 90%.Therefore,it is necessary to discover a new biomarker to improve the efficiency of diagnosis,treatment and prognosis of ovarian cancer.Zw10 binding factor,ZWINT,an important regulatory protein of chromosome movement and mitosis checkpoint,is attached between the chromosome and the microtubules of the spindle.ZWINT is overexpressed in a variety of human malignant tumors.Previous studies have reported that ZWINT was overexpressed in ovarian cancer,however,the effect and mechanism of ZWINT in ovarian cancer have not been elucidated.Therefore,in this study takes ZWINT as the core of the research to evaluate the clinical prospect of ZWINT as a target against epithelial ovarian cancer through clinical tissues.In addition,the influence of ZWINT on the phenotype of epithelial ovarian cancer from the cellular level were further explored to provide a theoretical basis for the treatment of ovarian cancer.Methods:1.Data analysis: Download the high-expressed genes in ovarian cancer from the c Bio Portal and GEO databases,the first 100 genes were screened separately,and after overlap,the published genes were excluded in the study to analyze the characteristics of the remaining genes in ovarian cancer.At the same time,the ovarian cancer tissue m RNA expression data and the survival data of matched patients were obtained from the TCGA database to analyze the relationship between target gene expression level and patient prognosis.2.Collected clinical samples and used immunohistochemical analysis of target gene expression in ovarian cancer tissues and normal ovarian tissues,and traced the relationship between target gene expression and the prognosis of the corresponding patients.3.Cell phenotype experiment: After the target gene is screened out,si-target gene was used to treat ovarian cancer cells(OVCAR3,A2780),the interference effect of target gene was verified by q PCR and WB experiments;the cell proliferation effect of target gene was detected by the experiments of cell colony formation,Ed U experiment and 6 days proliferation rate testes;in addition,flow cytometry was detected cell cycle to assess the potential of target gene to inhibit ovarian cancer;furthermore,cell starvation test was detected whether the migration and invasion of cells were affected by target gene.4.KEGG pathway analysis,target gene enrichment pathway,and verify the expression of related genes on this pathway.Results:1.ZWINT was screened out based on the data processing and analysis downloaded from the database,and the HR value of ZWINT was the most significant,which was closely related to the poor prognosis of patients with ovarian cancer.2.The results of immunohistochemistry showed that compared with normal ovarian tissue,the expression of ZWINT was significantly higher in ovarian cancer tissues;and the high expression of ZWINT was closely associated with the poor prognosis of ovarian cancer patients.3.After si-ZWINT treatment of ovarian cancer cell lines,the cell proliferation ability was significantly reduced.Enrichment analysis of KEGG pathway found that ZWINT was closely related to cell cycle pathways.After interfering with ZWINT,the cell cycle is inhibited in S phase,the proteins related to cell cycle(Cyclin D1,Cyclin B,Cyclin E1)were significant changes accordingly.4.The migration and invasion abilities of ovarian cancer cells after interference with ZWINT decreased significantly,and the proteins related to migration and invasion(Fibronectin,E-cadherin,N-cadherin,Vimentin)were changed significantly.Conclusion:1.ZWINT is significant highly expressed in ovarian cancer tissues and is closely related to the prognosis of patients with ovarian cancer;2.ZWINT mediates tumor cell phenotypes variation(proliferation,cycle,migration and invasion)of ovarian cancer cells;3.ZWINT may serve as a potential target of anti-epithelial ovarian cancer.