Effects On The Biological Behaviour And PI3k/AKT Signal Transduction Of VEGF、bFGF Which Induced By Estrogen Of Endometrial Cacirnoma Cell

Background: Endometrial cancer is one of the common gynecologicmalignancy, its incidence is on rising rencent years, but the exact cause isuncleared. Most scholars think that it may be related to the persistent stimulus ofendogenous and exogenous estrogen and lack of progesterone antagonist. Buthow does estrogen regulate cells proliferation, its mechanism is unknown. Ourprior studies have shown that treated with1x10-6mol/L estrogen for30min, thevascular endothelial growth factor (VEGF) and basic fibroblast growth factor(bFGF) mRNA and protein expression of the endometrial cancer cells weresignificantly increased. And the new blood vessel formation and cell adaptationto hypoxic environment is critical to the development of tumors. The occurringof tumor in volving multiple signal transduction pathways.Phosphatidylinositol-3kinase (P13K)-serine/threonine protein kinase B(PI3K/AKT) pathway is one of the most important signal transduction way tomediate membrane receptor signal transduction in the cells. PI3K/AKT can beactivated by a variety of growth factors and cytokines.Objective: To investigate the change of AKT, PI3K in PI3K/AKT pathway of endometrial cancer cells which were treated with E2and VEGF, bFGF inducedby estrogen. Research to estrogen stimulation of endometrial cancer cellIshikawa cells (estrogen receptor positive) and HEC-1A cells (estrogen receptorpositive frail), or give estrogen after seal off the membrane receptor of VEGF,bFGF.To find the changes of important protein in PI3K/AKT signaling. in orderto incestigate the relationship between estrogen，VEGF, bFGF (which inducedby estrogen) and PI3K/AKT signal transduction.Method：1. Real-time PCR and western blot were used to detect the concentration andtime of estrogen, LY294002, Bibf1120, Ponatinib. Investigate the expressionof VEGF, bFGF, PI3K, and AKT in Ishikawa cells and HEC-1A cells, thesecells were treated with1×10-6mol/L estrogen (E2group) for30min; or5010umol/L PI3K/AKT inhibitor LY294002for1h,1010umol/L VEGFreceptor inhibitor Bibf1120for1h,2.510umol/L bFGF receptor inhibitorPonatinib for1h, and then at a concentration1×10-6mol/L of estrogenincubated30min.2. MTT, FCM (Flow Cytometry Method) were used to investigate the cellsproliferation, cell cycle distribution.The metastasis capability of cells weredetected by trans-well assay. And the monoclonal ability was detected by cellcolony forming experiment.Result1. The AKT and PI3K in Ishikawa cells, HEC-1A cells activity peaked when1×10-6mol/L of E2stimulated30min. Inhibitory effect achieve the best thePI3K inhibitor LY294002, VEGF receptor inhibitor10ng/mL Bibf1120,bFGF receptor inhibitors2.5ng/ml Ponatinib in endometrial cancer Ishikawacells, HEC-1A cells one hour. Protein expression of VEGF、bFGF、PI3K、 AKT in the E2group were significantly higher than that in the control group(P<0.05).Compared with the estrogen group: the mRNA and proteinexpression of VEGF, bFGF, PI3K, AKT in LY294002+E2group were lower(p <0.05), Bibf112010+E2group, mRNA and protein expression of PI3K,AKT were decreased (p <0.05), Ponatinib+E2group, mRNA and proteinexpression of PI3K, AKT were decreased (p <0.05).2. Compared with the control group, when treated with1×10-6mol/L E230min,the proliferation of Ishikawa、HEC-1A cells is significantly increased（P＜0.05）, Trans-well assay showed that the migration and invasionability of cells is significantly boosted（P＜0.05）.Conclusion：Estrogen and VEGF、bFGF which were induced by estrogen ofIshikawa、HEC-1A cells, can significantly increase the proliferation, migration,invasion and cloning of the cells. They maybe through activating the PI3K/AKTsignal transduction pathway to play these biological roles.