The Effect Of O-GLcNAcylation On The Proliferation Of Ovarian Cancer Cells

Ovarian cancer is one of the malignant female tumors,which is a serious threat of women's health.Although ovarian cancer surgical procedure and postoperative treatments has been continuously improved,the five-year survival rate of ovarian cancer is only 44%.For the reason of the deficiency of early ovarian cancer diagnosis,a lot of patients have been found to be late or metastatic.At present,the common treatment for ovarian cancer is surgical and radiation and chemotherapy,but as a result of ovarian cancer cells resistance makes some patients has a poorer clinical treatment effect.In order to improve the survival rate and the prognosis of ovarian cancer patient,we need to find new targets for chemotherapy,which may improve the sensitivity of ovarian cancer cells to chemotherapy drugs.Objective: This study discusses the effect of O-GLcNAcylation on ovarian cancer cell proliferation through the following studies:Methods and Results: 1.Compare the O-GLcNAc expression in different ovarian cancer cell lines.We tested the expression of O-GLcNAc in SKOV3 cells,CAOV3 cells,OVCAR3 and A2780 cells by Western blot.The results showed that the O-GLcNAc expression is the highest in SKOV3 cells and the lowest in A2780 cells.2.Construct a lower expression and higher expression of O-GLcNAc ovarian cancer celllines.We used lentivirus to construct low expression of O-GLcNAc in SKOV3 cell and used PUGNAc to construct high expression of O-GLcNAc.The results of Western blot showed the two models were both effective.3.Explore the proliferation of ovarian cancer cell mediated by O-GLcNAc.We used flow cytometry technology to test cell cycle and CCK8 to test cell proliferation.The results showed that G1 phase was increased and G2 / M phase was decreased in low expression of O-GLcNA SKOV3 cells compared with control group.The results showed that G1 phase was decreased and G2 / M phase was increased in high expression of O-GLcNA A2780 cells compared with control group 4.O-GLcNAc modified the proliferation of the ovarian cancer cell through ?-catenin.We tested the expression of ?-catenin in the two pairs of cell models.The results shown that ?-catenin deceased in low expression of O-GLcNA SKOV3 cells and ?-catenin increased in high expression of O-GLcNA A2780 cells.But the mRNA level of ?-catenin shows no difference between the two groups.The results of Co-IP showed the modification of O-GLcNAc of ?-catenin was decreased in low expression of O-GLcNA SKOV3 cells and increased in high expression of O-GLcNA A2780 cells compared with control group.5.The elevated of ?-catenin mediated by O-GLcNAc translocation to the nucleus and trigger the downstream target genes.Finally,we tested the levels of downstream target genes of ?-catenin.And the results shown the levels of CyclinD1 and Axin2 was decreased in low expression of O-GLcNA SKOV3 cells and increased in high expression of O-GLcNA A2780 cells compared with control group.