| Background: Epithelial ovarian cancer(EOC)is the leading cause of death in gynecologic cancers with advanced disease in developed countries,The 5-year survival rate is about 20%.Although median survival has improved in most patients in recent years,but most patients eventually develop recurrence due to chemo-resistance and progress to progressive disease.Therefore,the treatment,diagnosis and prognosis of ovarian cancer,should be highly valued.Cancer antigen 125(CA125)is a repetitive peptide epitope of the tandem repeat domain of mucin 16(Mucin,MUC16),which has been used as a diagnostic marker for ovarian cancer at an early stage.extracellular domain of MUC16 extends from the cell surface in ovarian cancer cells and provides an early contact point for the metastatic ovarian cancer cells and distal peritoneal mesothelial cells.The intracellular domain of MUC16 may regulate the function of ovarian cancer cells by participating in cell transduction pathways.In this paper,the cell invasion assay found that CA125 has a significant impact on the migration of ovarian cancer cells and we made further exploration of the mechanism.The research provides an important reference for clinical diagnosis and treatment of ovarian cancer.Purpose: Chapter 1: We observe the effect of CA125 on the proliferation and migration of ovarian cancer cells.Chapter 2:We further explore the mechanism of CA125 affect the development of ovarian cancer and it’s related proteins or molecules.Chapter 3:We collect the Initial clinical serum levels of CA125 in patients with ovarian cancer after admission,and analyze the relationship between the level of serum CA125 and the progression of ovarian diseases.we try to prove the role of serum CA125 in the development of ovarian cancer and make a new critical value of CA125 for judging the metastasis of ovarian cancer.The research will provide the basis for the clinical treatment and prognosis of ovarian cancer.Methods: Chapter 1: The ovarian cancer cells were stimulated with different concentrations of MUC16 / CA125.MTT assay was used to observe the influence of MUC16 / CA125 on the proliferation of ovarian cancer cells and Transwell assay was used to confirm the impact of MUC16 / CA125 on the migration ability of ovarian cancer cells.Chapter 2:We observed changes in cell migration when the DKK1 and CA125 were acting on ovarian cancer cells simultaneously.Chapter 3: We treated ovarian cancer cell line with different concentrations of MUC16 / CA125 protein,and extracted the protein samples of stimulated ovarian cancer cells,the Western Blot experiments and Enzyme-Linked Immuno Sorbent Assay(ELISA)were used to find out the relatede pretein that impacted ovarian cancer cell migration.The total RNA of the stimulated ovarian cancer cells were ectracted and amplified by q PCR to find out more related proteins and molecules to further explore the signal pathway involved in the process.Chapter 4: The changes of cell migration were observed when DKK1 was over-expressed and silenced in ovarian cancer cells.Chapter 5:Collecting the information of patients with ovarian cancer like :the levels of serum CA125,cancer metastasis and ages of these patients,the receiver operating characteristic ROC curve was used to determine the relationship between the level of serum CA125 and cancer metastasis.A new threshold of CA125 level was found for the determination of ovarian cancer metastasis.Result: Chapter 1:Cell proliferation assay MTT found that when the ovarian cancer cells were stimulated by CA125 with the concentration of 0.2 ug / ml and 0.4 ug / ml for 48 hours,respectively,the proliferation of these cells was not significant.Transwell experiments were performed to find the change of migration ability in two ovarian cancer cells,and the migration of these cells was significantly enhanced after stimulating by CA125 for 48 hours,and the changes of migration ability in the two cell lines caused by CA125 were not exactly the same.Chapter 2: When DKK1 and CA125 were simultaneously acting on ovarian cancer cells,the DKK1 showed a suppressive effect on cell migration which was originally promoted by CA125.Chapter 3:The q PCR,Western Blot and ELISA showed that,the expression of DKK1 was significantly decreased in both ovarian cancer cell lines when stimulated with CA125 protein,and the effect was dependent on the concentration of CA125.Chapter 4:The ovarian cancer cells were selected from four ovarian cancer cell lines: A2780,OVCAR3,SKOV-3 and CAOV-3 for DKK1 overexpression,silencing,and cell migration experiments,we found the DKK1 can reverse the effect of CA125 on the migration of ovarian cancer cells.The result provides a powerful basis for the study of the mechanism of CA125 affecting the migration of ovarian cancer cells.Chapter 5: Collecting the initial information of patients,the serum CA125 levels,the age of patients and the tumor metastasis.ROC curve analysis was performed and found that serum CA125 level in patients with ovarian cancer can be used as a reference standard to judge the tumor metastasis,and the threshold is 82.9 U / ml,and the correlation analysis found that the age and serum CA125 levels of patients have important relevance with tumor metastasis of ovarian patients. |
