The Clinical Significance Of RAD51C Expression In Ovarian Cancer Tissue And Its Effect On Ovary Cancer Cells Of Proliferation, Migration And Apoptosis

Research background and objective:Ovarian cancer is one of the most common gynecological malignancies,with the highest mortality among them.Its morbidity increased every year,and morbidity crowd become younger gradually.At present,surgery combined with chemotherapy are the main treatment for ovarian cancer,and the relapse caused by chemotherapy drug resistance is one of the main causes of death,but the mechanism of drug resistance is still not very clear.Therefore,finding targeted drug therapy to reduce the recurrence has become the key to the treatment of ovarian cancer patients.Homologous recombination damage repair is related to the tumor development and prognosis,and also related to tolerance of tumor for radiation and chemotherapy.RAD51C belongs to the homologous recombination(HR)repair gene family and is one of the key proteins in the homologous recombination repair pathway,and its overexpression or mutation can changes the homologous recombination repair ability and cause genomic instability.The purpose of this study is to investigate the clinical significance of RAD51C expression in epithelial ovarian cancer tissue and its effect on the proliferation,migration and apoptosis of ovarian cancer cells.Methods:1.The expression of RAD51C in 82 cases of epithelial ovarian cancer and 80 cases of epithelial benign ovarian tumor was detected by immunohistochemical method,and its clinical significance was analyzed.2.The mRNA levels of RAD51C in ovarian cancer cell lines(SKOV3 cells,A2780 cells,CAOV3 cells)were detected by fluorescence quantitative PCR,and the highly expressed cell lines were selected as the research objects in the later stage.3.Three interference vectors were designed for the target cells.The expression of RAD51C in the target ovarian cancer cells after transfection was verified by qPCR and western-blot,and the cell line with the best interference effect was selected as the later research object.4.Cell proliferation capacity of normal control group,no-load group and interference group was detected by CCK8 method.5.The ability of cell migration and infiltration in normal control group,no-load group and interference group was detected by scratch test.6.Cell apoptosis was detected by flow cytometry in the normal control group,the no-load group and the interference group.Results:1.The expression of RAD51C protein in ovarian cancer tissues was higher than that in benign ovarian cancer tissues(P<0.01),with a statistically significant difference.Moreover,the expression of RAD51C protein in ovarian cancer tissues was related to the patient's age,pathological type,clinical stage and prognosis,with a statistically significant difference(P<0.05).2.Compared with SKOV3 cells and CAOV3 cells,the mRNA level of RAD51C in A2780 cells was significantly higher than that in the control group(P < 0.05).Therefore,A2780 cell line was selected as the research object in the later stage.3.Three RAD51C interference carriers were transfected to A2780 cells,and then the mRNA expression was detected by qPCR,protein expression changes was test by Western Blot.In A2780 cells,the RAD51C interference 3 worked best.Comparing with the control group,RAD51C mRNA and protein expression level decreased significantly(P< 0.05),with a statistically significant difference.Thus,the siRNA-3was chosen as the later research object.4.Interference with RAD51C gene detected by CCK8 significantly reduced cell proliferation,and the difference was statistically significant(P< 0.05).5.Interference with RAD51C gene in cell scratch test could reduce the migration ability of cells.After 24 h,the migration ability of cells in si-RAD51C group was significantly lower than that in the control group,and the difference was statistically significant(P < 0.05).6.Interference with RAD51C gene was detected by flow cytometry to promote apoptosis,especially at the early stage of apoptosis.The difference was statistically significant(P < 0.05).Conclusion:Compared with epithelial ovarian benign tumors,RAD51C protein in epithelial ovarian cancer tissues significantly higher expression,which is significantily correlated with the patient's age,tumor nature,FIGO clinical stage,pathological type and the prognosis of patients in ovarian cancer tissue,especially in the high grade serous ovarian cancer.Knocking down the RAD51C gene in human ovarian cancer cell line can significantly inhibit the proliferation and migration of ovarian cancer cells and promote apoptosis.RAD51C may be involved in the development of ovarian cancer,and may be a target for the detection of target genes and targeted therapy of ovarian cancer.