Effect Of FBXW7 On The Growth Of Epithelial Ovarian Cancer And Its Mechanism

ObjectiveMalignant neoplasms are a serious threat to human health,and ovarian cancer is the fifth largest tumor in women's cancer-related deaths that seriously threatens women's life and health.Therefore,the study of important oncogene and tumor suppressor gene function not only provides the basis for the risk assessment and personalized treatment of ovarian cancer,but also provides a new direction for finding new targets for cancer targeting and new targeted anticancer drugs.FBXW7 is an E3 ubiquitin ligase that is involved in the regulation of Aurora-A,Notch,c-Jun and other protein and signaling pathways in a variety of human tumors such as cholangiocarcinoma,leukemia,endometrial cancer and breast cancer.In the process of tumor development play a role in tumor suppressor gene,but FBXW7 in Epithelial Ovarian Canceroccurrence and development of the role and mechanism is not clear,and without literature reported.This study will focus on the expression of FBXW7 in Epithelial Ovarian Cancerand its function in the regulation of Epithelial Ovarian Cancercells and subcutaneous xenograft tumor growth and mechanism.MethodsA2780s,A2780 s,SKOV3 and CAOV3 Epithelial Ovarian Cancercells and normal ovarian epithelial cells HOEC were culture and WB and real-time PCR were performed to detect FBXW7 gene expression at mRNA and protein levels.On this basis,we constructed pcDNA3.1 vector-mediated human FBXW7 overexpression plasmid and then used to transfect A2780 cp and A2780 s cells.In the use of WB clear p-FBXW7 can effectively promote the expression of FBXW7 protein in Epithelial Ovarian Cancercells,we will use the CCK8 test to detect cell proliferation at the cellular level,the clone formation assay was used to detect the cell clone formation ability,and the cells were analyzed by flow cytometry apoptotic and cell cycle distribution.Based on the FBXW7 regulation of the biological characteristics of Epithelial Ovarian Cancercells,we used WB to detect the expression of apoptosis-related protein Bcl-2,Bax,Bcl-xL,Caspase-3 and cell cycle-related protein Cyclin B1,Cyclin D1 and Cyclin E1 expression in FBXW7 overexpressing Epithelial Ovarian Cancercells and control cells.On the basis of the study of cell level,we screened the obtained FBXW7 stable expression of A2780 cp cells and control cells inoculated into the right ventral subcutaneous nude mice to establish subcutaneous xenografts,and then draw the tumor growth curve and weighed mice tumor weight.Finally,we will peel the transplanted tumor tissue of mice,and then employ immunohistochemical staining to detect the expression of FBXW7,proliferation-related indicators of PCNA and angiogenesis markerCD31.The apoptotic cells were detected by TUNEL assay.ResultsWB and real-time PCR results showed that FBXW7 was low or no expression in Epithelial Ovarian Cancercells compared with normal ovarian epithelial cell HOEC.Overexpression of FBXW7 in epithelial ovarian cancercells A2780 cp and A2780 s cells can significantly inhibit cell proliferation and clonal formation,promote cell apoptosis and block cell cycle.Further molecular mechanism studies showed that FBXW7 could significantly promote the expression of Bax and Caspase-3,inhibit the expression of Bcl-2 and Bcl-xL,and inhibit the expression of Cyclin E1 and Cyclin D1,but there was no significant regulation of Cyclin B1 expression.In the study of animal level,we found that FBXW7 significantly inhibited the tumor volume of A2780 cp cells subcutaneously transplanted tumor(inhibition rate was 70.6%;Ctrl group tumor volume: 1463.2 ± 175.3mm3;FBXW7 group tumor volume: 432.5 ± 65.3mm3,Figure 12B)and tumor tumor(inhibition rate of 69.5%;Ctrl group tumor volume: 1.38 ± 0.21g;FBXW7 group tumor volume: 0.42 ± 0.08 g,Figure 12C).Immunohistochemical staining showed that overexpression of FBXW7 in tumor tissue could significantly inhibit cell proliferation and angiogenesis and promote apoptosis.ConclusionFBXW7 is low expressed in epithelial ovarian cancercells.Overexpression of FBXW7 can significantly inhibit cell proliferation and clonal formation by promoting cell apoptosis and inhibiting cell cycle progression.Overexpression of FBXW7 in subcutaneous transplanted epithelial ovarian cancercan significantly inhibit the growth of subcutaneous xenografts by significantly inhibiting tumor tissue proliferation,angiogenesis and promoting apoptosis.The above studies have shown that FBXW7 plays an important role in the development and progression of epithelial ovarian cancerand is a potential target for the diagnosis and treatment of ovarian cancer.