Fsh Regulated The Molecular Mechainsm Of Vegf In Epithelium Ovarian Cancer Via The Microrna-27A Pathway

Epithelial ovarian cancer is taking the primary mortality in female reproductive tract tumor,which is the main tumor threating the health of women in our country. In human body, FSH is closely related to the development of epithelial ovarian cancer. It can not only promote to the proliferation, invasion and inhibit the apoptosis,but also can induce to the inflammation,survival and angiogenesis via regulating COX-2,Survivin,VEGF,VEGFR1/Flt-1.Our research group has the leading levels in the area of FSH and ovarian cancer.we had proved the relationship between FSH and COX-2,Survivin,VEGF in epithelial ovarian cancer cells.COX-2and Survivin are synergetic proteins, so they are written with COX-2/Survivin; FSH can promote the expression of COX-2/Survivin and VEGF; FSH can upregulate the expression of VEGF through PI3K/AKT-Survivin and upregulateing HIF-1a pathways. Howere,the Nowadays, the molecular mechanism of FSH inducing to COX-2,VEGF is still unknown,so it is becoming one of the hot researchs in Gynecologic cancers.In recent years,the studying about miRNAs are extremely prevalence and some hormone have found making effects on miRNAs.There is only few articles reported the relation between FSH and miRNAs,even the research about FSH and miR-27a have no yet been reported.In addition,there are only three articles reported the FSH can regulate the expression of Survivin and VEGF through the miR-27a:ZBTB10-SP1signaling pathway.Therefore,if FSH can regulate the expression of COX-2/Survivin,VEGF via the G-protein undependent way of miR-27a:ZBTB10-SP1, there is no demonstration in domestic or abroadObjective:To explore the effect of follicle-stimulating hormone on the expression of miR-27a gene,ZBTB10,SP1,COX-2/Survivin,VEGF proteins in epithelium ovarian cancer cells, and analysis the relationships among those molecule. Further study whether the effect is induced by microRNA-27a:ZBTB10-SP1signaling way and regulates the expression of inflammatory,Survival,vascular factors, including COX-2/Survivin,VEGF in epithelium ovarian cancer.Methods:1. In this study, the genes were inhibited or overexpressed in the hypothetical signaling pathway. In order to detect the relationships to be upstream and downstream among those molecule, we used the techniques of TaqMan real-time PCR,immunochemistry,Western blot to verify the handing factors taken effects on the other genes or proteins. Further study whether FSH can regulate the expression of inflammatory factor, survival factor, vascular factor,including COX-2/Survivin,VEGF via the miR-27a:ZBTB10-SP1signaling pathway.2. Statistical analysis:the result of real-time PCR and grey scale of Western blotting were denoted as x±s. One way ANOVA were used in spss17.0. p<0.05was significant and considered significant at P<0.01in each comparison. Results:1. The expression of microRNA-27a was higher on five epithelium ovarian cancer cells than normal epithelium ovarian cell,except the A2780;the expression of SP1,ZBTB10were different in six epithelium ovarian cancer cells; The expression of SP1is higher than ZBTB10in epithelium ovarian cancer tissues.2. Taking the HEY and HO8910for representative,the follicle-stimulating hormone regulated the expression of microRNA-27a and the protein of SP1,ZBTB10in a dose-dependent manner and significant influence occoured when the follicle-stimulating hormone concentration was50mIU/ml.Follicle-stimulating hormone up-regulated the expression of microRNA-27a and SP1,while down-regulated on ZBTB10;The follicle-stimulating hormone up-regulated the expression of COX-2/Survivin,VEGF. HEY and HO8910are choosing as the following experiment,the working concentration of follicle-stimulating hormone was50mIU/ml.3. The cells were treated with anti-microRNA-27a,PZBTB10,SiSPl which could inhibit the expression of COX-2/Survivin,VEGF treated by FSH. Meanwhile,the expression of ZBTB10was increased and SP1was decreased after adding the anti-microRNA-27a,which was also hinder the effecting that FSH taking on ZBTB10and SP1;The expression of SP1was decreased after adding the PZBTB10,which was also hinder the effecting that FSH taking on SP1;The activity of COX-2/Survivin,VEGF were inhibited after adding the siSPl. Conclusion:follicle-stimulating hormone enhance the activity of inflammation and vascularization through provoking microRNA-27a:ZBTB10-SP1signaling way.