TLR4 Signaling Pathway Activation Up-regulates The Expression Of KDM2B And Influences The Proliferation And Migration Of Ovarian Cancer Cells

ObjectiveTo use STRING and GEPIA databases to predict the interaction between TLR4 and KDM2 B.To detect the expression of Toll-like receptor 4(TLR4)and histone demethylase KDM2 B in normal ovarian tissues,benign ovarian tumors and ovarian malignant tumors situation,analyze the relationship between the two and the clinical pathological characteristics of ovarian cancer and their clinical significance,and clarify the correlation between the two.Study the effect of TLR4 pathway activation on the expression of KDM2 B in ovarian cancer cells SKOV3 and A2780 cell lines,and the effects of biological behavior of ovarian cancer cells.Methods1.Use STRING to analyze the protein-protein interaction network(PPI)of TLR4 and KDM2 B,and analyze the correlation between TLR4 and KDM2 B in ovarian cancer in GEPIA.2.Immunohistochemistry was used to detect the expression of TLR4 and KDM2 B protein in normal ovarian,benign ovarian tumors and ovarian cancer tissues,and to analyze the expression of TLR4 and KDM2 B protein in ovarian cancer tissues and the clinical pathological characteristics of ovarian cancer Correlation.3.Cell immunofluorescence assay was used to detect the expression of TLR4 in human ovarian cancer cell lines SKOV3 and A2780.Cell Counting Kit-8 was used to detect the 24-hour cell proliferation of ovarian cancer cell lines SKOV3 and A2780 cells stimulated by different concentrations of TLR4 ligand lipopolysaccharide(LPS),and the optimal LPS concentration for enhancing the proliferation ability of ovarian cancer cells was determined.LPS was used to activate TLR4 Signaling pathway,antagonistic TLR4 receptor pretreatment LPS activates TLR4 signaling pathway,using real-time fluorescent quantitative PCR and Western blot to detect TLR4 and KDM2 B m RNA and protein expression in ovarian cancer cell lines SKOV3 and A2780.Cell Counting Kit-8 was used to detect the proliferation of ovarian cancer cells,and Transwell assay was used to detect the migration of ovarian cancer cells.Results1.STRING line protein-protein interaction network(PPI)analysis results show that there may be an interaction relationship between TLR4 and KDM2 B.Analysis results in GEPIA show that TLR4 and KDM2 B are positively correlated in ovarian cancer.2.The positive expression of TLR4 and KDM2 B protein in epithelial ovarian cancer tissue are significantly higher than that of benign ovarian tumors and normal ovarian tissue(P<0.05).The expression of TLR4 and KDM2 B protein in ovarian cancer is related to tumor tissue grade,FIGO stage and lymph node metastasis,and the expression of the two in ovarian cancer tissue is positively correlated(P<0.05).3.Cell immunofluorescence assay confirmed that TLR4 is highly expressed in human ovarian cancer cell lines SKOV3 and A2780.The detection results of Cell Counting Kit-8 showed that(1,5,10,20,40ug/ml)LPS stimulated ovarian cancer A2780 cells to increase cell proliferation ability(P<0.05),in which the LPS concentration was 20 ug /ml A2780 cells The most proliferative ability(P<0.05).In SKOV3 cell lines,the concentration of 10ug/ml and 20ug/ml LPS stimulated ovarian cancer SKOV3 cells to increase the proliferative ability(P<0.05).while 10ug/ml and 20ug/ml,the proliferative capacity of SKOV3 cells was not statistically significant(P>0.05).When LPS activates the TLR4 signaling pathway,the m RNA and protein expressions of TLR4 and KDM2 B are increasing(P<0.05).TAK-242 antagonizes TLR4 receptor pretreatment before using LPS to activate the TLR4 signaling pathway,TLR4 and KDM2 B m RNA and protein expression The amount was reduced when LPS alone was used to activate the TLR4 signaling pathway(P<0.05).When LPS activates TLR4 signaling pathway,the proliferation and migration ability of ovarian cancer cells is enhanced(P<0.05).TAK-242 antagonizes TLR4 receptor pretreatment and then uses LPS to activate TLR4 signaling pathway.LPS alone attenuates TLR4 signaling pathway(P <0.05).Conclusion1.The high expression of TLR4 and KDM2 B protein in epithelial ovarian cancer tissues is related to tumor tissue grade,FIGO staging and lymph node metastasis.The expression of the two in ovarian cancer tissues is positively correlated.2.In ovarian cancer A2780/SKOV3 cells,activation of TLR4 signaling pathway up-regulates the expression of KDM2 B and promotes the proliferation and migration of ovarian cancer cells.3.In ovarian cancer A2780/SKOV3 cells,TLR4 expression increased after activating TLR4 signaling pathway.