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目的 探讨 17β 雌二醇 (E2 )对子宫内膜癌细胞系Ishikawa细胞磷脂酰肌醇 3激酶 /蛋白激酶B(PI 3K/PKB)信号传导通路的激活作用以及PI 3K抑制剂对其的影响。方法 应用免疫印迹杂交技术 ,检测一定浓度 (1× 10 -6mol/L) 17β E2 作用于Ishikawa细胞不同时间 (分别为 0、5、15、30、6 0、12 0min)后和不同浓度 (分别为 0、1× 10 -10 、1× 10 -8、1× 10 -6、1× 10 -4mol/L) 17β E2 作用Ishikawa细胞一定时间 (30min)后PKB的活化情况 [以磷酸化PKB和总PKB比值 (p PKB/PKB)表示 ],以及用同法检测PI 3K抑制剂LY2 94 0 0 2对 17β E2 活化PKB的影响。结果 1× 10 -6mol/L 17β E2 作用于Ishikawa细胞 15min时 ,PKB的活化水平 (0 5 33± 0 0 2 9)已明显高于对照 (0 36 1± 0 0 2 9,P <0 0 5 ) ,30min时最明显 (0 6 6 6± 0 0 2 1,P <0 0 0 1) ,而且持续至少达 12 0min时 ;随着 17β E2 浓度 (分别为 1× 10 -10 、1×10 -8、1× 10 -6、1× 10 -4mol/L)的增加 ,PKB的活化水平逐渐增高 ,与对照比较 ,差异有显著性 (除 1×10 -10 mol/L外 ,P值分别为 >0 0 5、<0 0 5、<0 0 5、<0 0 0 1)。随着LY2 94 0 0 2作用浓度 (分别为 0 1、10、5 0、10 0 μmol/L)的增加 ,17β E2 作用Ishikawa细胞后P
Objective To investigate the activation of PI3K / PKB signal transduction pathway by 17β estradiol (E2) on Ishikawa cells and the effect of PI 3K inhibitor on it. Methods Western blotting was used to detect the effect of 17β E2 at a certain concentration (1 × 10 -6 mol / L) on Ishikawa cells at different times (0, 5, 15, 30, 60, 120 min, (0, 1 × 10 -10, 1 × 10 -8, 1 × 10 -6, 1 × 10 -4 mol / L) 17β E2 activation of Ishikawa cells after a certain time (30min) PKB activation [phosphorylated PKB and Total PKB ratio (p PKB / PKB)] and the effect of the PI 3K inhibitor LY2 94 0 0 2 on 17β E2-activated PKB by the same method. Results The activation level of PKB (0 5 33 ± 0 0 2 9) in Ishikawa cells treated with 1 × 10 -6 mol / L 17β E2 for 15 min was significantly higher than that of the control (0 36 1 ± 0 0 2 9, P 0 0 0 5), the most obvious at 30min (0 6 6 6 ± 0 0 2 1, P 0 01), and continued for at least 12 0min; with 17β E2 concentration (respectively 1 × 10 -10, 1 × 10 -8,1 × 10 -6, 1 × 10 -4 mol / L) increased the activation level of PKB gradually increased compared with the control, the difference was significant (except 1 × 10 -10 mol / L, P value Respectively> 0 0 5, <0 0 5, <0 0 5, <0 0 0 1). With the increasing concentration of LY2 94 0 0 2 (0 1, 10, 50 and 100 μmol / L, respectively), 17β E2 treated Ishikawa cells with P