目的 观察磷脂酰肌醇-3-激酶(P13K)催化亚基p110及磷酸化蛋白激酶B(p-Akt)、蛋白激酶B(Akt)在体外培养血管瘤内皮细胞中的表达,探讨其与血管瘤内皮细胞周期分布和凋亡的关系.方法 用组织块法培养血管瘤内皮细胞,同步化培养血管瘤内皮细胞后,采用Western blot法检测血管瘤内皮细胞PI3Kp110、p-Akt及Akt蛋白的表达水平,同期用流式细胞仪检测血管瘤内皮细胞周期及凋亡的变化,并分析P13Kp110、p-Akt及Akt蛋白的表达水平与血管瘤内皮细胞周期分布与凋亡的相关性.结果 p110、p-Akt蛋白的表达水平低时(0.19±0.01、0.09 ±0.02),Go/G1期细胞比例为(94.00±3.00)％,细胞总凋亡率为(11.24±2.34)％,p110、p-Akt蛋白的表达水平高时(0.37±0.04、0.22 ±0.01),Go/G1期细胞比例为(61.00±6.00)％,细胞总凋亡率为(0.51±0.03)％,二者比较差异均有统计学意义(t=-8.42、-35.48,P均＜0.01),而总Akt蛋白表达不变(0.72±0.00).结论 体外培养的血管瘤内皮细胞中有p110、p-Akt及Akt蛋白的表达.p110、p-Akt参与体外培养的血管瘤内皮细胞凋亡的调节.%Objective To study the expression of phosphatidy linositol-3-kinase(PI3K) p110 catalytic subunit and phosphorylation-protein kinase B(p-Akt),protein kinase B(Akt) in cultured vascular endothelial cells,then the level of PI3K p110,p-Akt and Akt expression and the correlation with hemangioma vascular endothelial cell cycle and apoptosis were analyzed.Methods The hemangioma vascular endothelial cells which came from the sterile hemangioma were cultured by using tissue culture method and then synchronized cultured endothelial cells,P13K p110,p-Akt and Akt of them were detected by way of Western blot,meanwhile cell cycle and cell apoptosis were examined by using flow cytometry technique,then the level of PI3K p110,p-Akt and Akt expression and the correlation with hemangioma vascular endothelial cell cycle and apoptosis were analyzed.Results The percentage of vascular endothelial cell was (94.00 ± 3.00) ％ in the G0/G1 phase and the apoptosis rate was (11.24 ± 2.34) ％ when p110 and p-Akt protein expression level were lower (0.19 ±0.01,0.09 ±0.02),and they were (61.00 ±6.00)％ and (0.51 ±0.03)％ when p110 and p-Akt protein expression level were higher (0.37 ±0.04,0.22 ±0.01).The percentage of vascular endothelial cell in the G0/G1 phase and the apoptosis rate were different in 2 groups(t =-8.42,-35.48,all P ＜0.01).But Akt protein expression level was not changed (0.72 ± 0.00).Conclusions p110,p-Akt and Akt protein were expressed in the cultured hemangioma vascular endothelial cells in vitro.PI3K p110,p-Akt may be involved in cultured hemangioma vascular endothelial cells apoptosis in vitro.
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