The Experimental Study On The Mechanism Of EGF/FOXM1 In Ovarian Cancer Metastasis

Objectives: The ovarian cancer occurred with the interaction of various genes pathways, the study of the function of these genes contributed to the gene-targeted cancer therapy. EGF, FOXM1 gene indicated that they may be involve in the occurrence and malignant progression of ovarian cancer, the subject is to study the action and mechanism of EGF / FOXM1 in ovarian cancer cells proliferation, metastasis by clinical specimens and in vitro test.Methods: The expressions of FOXM1 in ovarian cancer tissues specimens collected clinically and normal ovarian tissue specimens were detected by immunohistochemical SP method; the expressions of FOXM1 in human ovarian cancer cell HO8910 PM, OVCAR3, SKOV3 and other cells were analyzed by Western blot; after different concentrations of EGF acting on SKOV3 cells, the expressions of FOXM1 were detected by Western blot; the effect of different concentrations of EGF on SKOV3 cells proliferation were detected by cck8; the effects of different concentrations of EGF on invasion of SKOV3 cells were detected by cell scratch test and Transwell chamber method; design and synthesize three si RNA expression sequences of targeted human FOXM1 gene. Total proteins were extracted after co-transfection liposome LipofectamineTM2000 to SKOV3 cells for 24 h. FOXM1 protein expressions in SKOV3 cells after transfection were analyzed by the application of Western blot, select and knockout the best effectiveness of si RNA sequences; the proliferation inhibition rates of SKOV3 cells were detected by cck8 after knockout FOXM1.Results: 1) FOXM1 expression in ovarian cancer tissues was significantly higher than that in normal ovarian tissue, FOXM1 expressed significantly in OVCAR3 cells, SKOV3 cells, HO8910 cells of ovarian cancer, in which expression of SKOV3 cells was higher than the other two cells; 2) After treating with different concentrations [concentrations(ng / ml) = 0,5,10,25,50,100] of EGF, SKOV3 cells underwent Western blot assay indicated that FOXM1 had highest expression when concentration of EGF was 10 ng / ml(P <0.05); 3) After treating with different concentrations [concentrations(ng / ml) = 0,5,10,25,50,100] of EGF, SKOV3 cells underwent cck8 assay indicated that there was highest absorbance of cells when the concentration was 10 ng / ml(P <0.01); EGF concentration of 10 ng / ml acted on SKOV3 cells as the study group, without EGF as control group, After 24 h Transwell cell migration assay, electron micrography indicated that the cell migration rate of study group than that of control group(P = 0.002), after 24 h invasion assay, electron micrography indicated that the cell migration rate of the study group was greater than that of the control group(P = 0.015); 4) after three targeted FOXM1 of si RNA infecting SKOV3 cells, Western blot confirmed that the silence effect of si RNA-FOXM1-801 was best; cck8 assay confirmed that after si RNA-FOXM1-801 silencing FOXM1,it can inhibit SKOV3 cell proliferation significantly(P = 0.005); cck8 assay confirmed after FOXM1 silencing by si RNA-FOXM1-801, statistical results of absorbance had no significant difference between the control group(not plus EGF) and the study group(EGF = 10 ng / ml)( P = 0.882).Conclusions: FOXM1 gene over expressed in ovarian carcinoma tissues, FOXM1 over expressing was in three ovarian cancer cell lines with the highest expressions of SKOV3 cells; promotion of expressing FOXM1 in SKOV3 cells most obvious when EGF concentration was 10ng/ml and enhanced proliferation and invasion of SKOV3 cells, EGF losing effect on cell proliferation after knockout FOXM1 gene, thus proving EGF may have an indirect regulating effect on ovarian cancer cells proliferation and metastasis through EGF / FOXM1 pathways.