The Effects Of CircN4BP2L2 On Biological Behavior Of Ovarian Cancer And Differential Phosphoproteomics Analysis Of Ovarian Cancer

Part 1 The effects of CircN4BP2L2 on BiologicalBehavior of Ovarian CancerBackground&Objective:Ovarian cancer is the most lethal gynecologic malignancy,accounting for 5%of female cancer-related deaths.Epithelial ovarian cancer is the most common histological subtype,accounting for 90%of all ovarian cancer cases.Due to lacking of early detection method and limited treatments,the 5-year survival rate is only 30%.Exploring the pathogenesis of ovarian cancer and finding potential diagnostic and therapeutic targets are of great significance to improve the 5-year survival rate.More and more studies have shown that circRNAs plays an important role in cancer and is a potential biomarker and therapeutic target.However,the expression and function of circRNAs in ovarian cancer are still unclear.In our previous study,we found that the expression of circN4BP2L2 was significantly down-regulated in epithelial ovarian cancer tissues by circRNAs sequencing technique.Low level of circN4BP2L2 was associated with advanced FIGO stage and lymph node metastasis.However,the mechanism of circN4BP2L2 in ovarian cancer is still not clear.In this study,we explored the effects of circN4BP2L2 on the biological behaviors of ovarian cancer SKOV3 cells,so as to provide reliable experimental and theoretical basis for the specific mechanism.Methods:CircN4BP2L2-siRNA was synthesized in vitro and transiently transfected into SKOV3 cells.We used qRT-PCR to detect the expression of circN4BP2L2 after transfection.The proliferation of SKOV3 cells was examined by CCK-8 assay and clonogenic assay.The migration of SKOV3 cells was detected by wound healing assay and Transwell assay.The invasion of these cells was detected by Transwell assay.Results:1.After transfection with circN4BP2L2-siRNA,the expression of circN4BP2L2 in the circN4BP2L2-siRNA group was down-regulated significantly compared with the negative control group(P<0.05).2.The migration and invasion of the group transfected by circN4BP2L2-siRNA was significantly enhanced(P<0.01).Conclusions:1.RNA interference technique can significantly inhibit the expression of circN4BP2L2 in ovarian cancer SKOV3 cells.2.CircN4BP2L2 can inhibit the migration and invasion of ovarian cancer SKOV3 cells and play the role of tumor suppressor genes in ovarian cancer.Part 2 Differential Phosphoproteomics Analysis of Ovarian CancerBackground&Objective:Ovarian cancer is one of the top ten cancers with the highest mortality in the world.Due to the lack of effective screening methods,the majority of patients are diagnosed in the advanced stage,causing chemoresistance and tumor recurrence.The conventional treatment is still a combination of cytoreductive surgery and platinum-based chemotherapy.Exploring its pathogenesis and finding potential therapeutic targets are very important to improve the prognosis of patients.In recent years,proteomics and phosphoproteomics have become an important tool in tumor research.In this study,we used quantitative phosphoproteomics method to comprehensively analyze the phosphoproteomic characterization for ovarian cancer.We explored the key phosphoproteins,and the corresponding cancer-related metabolic and signal pathways.Methods:We took advantage of a quantitative phosphoproteomics method on ovarian tissue samples obtained from 5 ovarian cancer patients and 5 matched controls.Results:A total of 722 phosphorylated sites corresponding to 534 proteins were significantly different(fold change≥2,p<0.01)between ovarian cancer patients and the controls.Among them,65 transcription factors and 29 kinases were included.Further functional analysis suggested fructose and mannose metabolism was highly enriched and involved in the pathogenesis of ovarian cancer.Four glycolytic enzymes(PFKFB2,PFKL,ALDOA,and TPI1)mapped to fructose and mannose metabolism were significantly changed in ovarian cancer.Moreover,the regulatory roles of modulated pathways,including MAPK,ErbB,and GnRH signaling pathways were also identified as critical processes involved in ovarian cancer.Conclusions:These findings may not only provide potential biomarkers and novel therapeutic targets for ovarian cancer,but also bolstered our knowledge on the physiopathology of this syndrome.