中文摘要：

目的：探讨雌二醇（E2）诱导子宫内膜癌Ishikawa细胞生成的血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）对磷脂酰肌醇3激酶/蛋白激酶B（PI3K/AKT）通路的主要基因蛋白的影响.方法：（1）以不同浓度E2作用于Ishikawa细胞30 min后,采用蛋白印迹法检测p-AKT蛋白水平.（2）实验设3组：①对照组;②E2组;③抑制剂＋E2组（包括BIBF1 120＋ E2组、Ponatinib＋ E2组和LY294002＋ E2组）,用荧光定量PCR和蛋白印迹法分别检测子宫内膜癌细胞内VEGF、bFGF、PI3K、AKT基因及蛋白的表达,流式细胞仪检测细胞周期的变化,Transwell检测细胞迁移能力.结果：（1）不同浓度（分别为0.01,0.1,1,10,100 μmol/L）的E2作用30 min后Ishikawa细胞中p-AKT蛋白的表达水平均高于对照组（P＜0.05）,且E2浓度为1 μmol/L时其表达水平最高;（2）E2组中VEGF、bFGF、PI3K、AKT mRNA及VEGF、bFGF、p-PI3K、p-AKT蛋白活化水平均高于对照组（均P ＜0.05）.BIBF1120＋ E2组、Ponatinib＋ E2组、LY294002＋ E2组细胞中PI3K、AKT基因及p-PI3K、p-AKT蛋白的表达均低于E2组（均P＜0.05）;（3）E2组与对照组相比,G1期细胞比例减少,S期细胞比例增加（均P＜0.05）,迁移穿过微孔的细胞增加（均P＜0.05）.LY294002＋E2组、BIBF1120＋E2组、Ponatinib＋ E2组分别与E2组相比,G1期细胞比例增加,S期细胞比例减少（均P＜0.05）,迁移穿过微孔的细胞减少（均P＜0.05）.结论：E2通过非基因转录效应产生VEGF、bFGF可以激活PI3K/AKT通路,促进子宫内膜癌的发生和转移.

英文摘要：

Objective： To investigate the effects of VEGF and bFGF induced by estrogen on PI3K/AKT pathway of endometrial cancer Ishikawa cells. Methods： （1）Western blot was used to detect the expression of phosphorylation AKT （p-AKT） protein in Ishikawa cell after stimulated with different concentrations of estradiol （0.01,0.1,1,10,100 μmol/L） for 30 mim. （2） There were 3 groups in the experiment：（1）control group;（2）E2 group; （3） Inhibitor＋ E2 group： including BIBFll20 ＋ E2 group, Ponatinib ＋ E2 group and LY294002＋ E2 group. And then mRNA level of VEGF,bFGF,PI3K,AKT and its relative protein respectively detected by qRT-PCR and western blot. Flow cytometry was used to examine the change of cell cycle, and transwell assay was used to detect the cell migration in different groups. Results： （1）The expression of the p-AKT protein at 0. 01,0.1,1,10,100 μmol/L E2 were significantly higher than that of control group, and when the concentration of E2 was 1 μmol/L, the expression of p-AKT was highest（ P 〈0.05）.（2） The mRNA levels of VEGF, bFGF, PI3K, AKT and the protein of VEGF,p-PI3K,p-AKT in E2 group were higher than that of control group（ P〈0.05）. The mRNA of PI3K and AKT ,and the protein of p- PI3K,p-AKT in BIBF1120＋E2 group,Ponatinib＋E2 group and LY294002-t-E2 group were together lowerthan that of Ez group（ P d0.05）. （3）Compared with the control group, the results of cell cycle of E2 group showed that percentage of G1 phase was decrease and S phase was significantly increase （ P〈 0.05）. But when compared with E2 group, opposite result showed in LY294002 ＋ E2 group, BIBF1120 ＋ E2 group or ponatinib＋E2group （P 〈0. 05）. Transwell assay also showed that the number of cell migration in E2 group was higher than that of control group but significantly lower than that of LY294002＋E2 group, BIBF1120＋E2 group or Ponatinib＋E2 group （ P 〈0.05）. Conclusion： VEGF and bFGF induced by E2 can active PI3K/AKT pathway ,and then promote