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目的:探讨低氧环境下磷脂酰肌醇3激酶(phosphatidylinositide3kinase,PI3K)抑制剂LY294002对人胶质瘤U251细胞中PI3K/丝氨酸-苏氨酸激酶(serine-threonine kinase,Akt)信号通路下游缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)mRNA和蛋白的表达以及细胞生长、细胞凋亡、细胞黏附和侵袭能力的影响。方法:应用LY294002作用于低氧环境中的人胶质瘤U251细胞(低氧+LY294002组),以低氧环境中未加LY294002的细胞作为对照(低氧组)。CCK-8试剂盒检测U251细胞生长,FCM检测细胞凋亡百分比,黏附实验检测细胞黏附能力,侵袭实验检测细胞侵袭能力,蛋白质印迹法检测p-Akt蛋白的表达,RT-PCR和蛋白质印迹法检测HIF-1αmRNA及蛋白的表达。结果:LY294002能明显抑制p-Akt蛋白以及HIF-1αmRNA和蛋白的表达,与低氧组比较差异均有统计学意义(P<0.05)。LY294002能抑制低氧环境中U251细胞的增殖,抑制效果在药物作用后第2～6天时最为明显(P<0.05)。低氧+LY294002组中细胞凋亡百分比明显高于低氧组(P<0.01),而细胞黏附和侵袭能力明显低于低氧组(P<0.01)。结论:LY294002能抑制低氧环境中PI3K/Akt信号通路下游HIF-1αmRNA及蛋白的表达,抑制人胶质瘤U251细胞的生长,促进细胞凋亡,并抑制细胞黏附和侵袭能力。
AIM: To investigate the effect of LY294002, a phosphatidylinositide 3 kinase (PI3K) inhibitor, on hypoxia of PI3K / serine-threonine kinase (Akt) signaling pathway in human glioma U251 cells under hypoxia Effects of hypoxia inducible factor-1α (HIF-1α) mRNA and protein expression and cell growth, apoptosis, cell adhesion and invasion ability. Methods: Human glioma U251 cells (hypoxia + LY294002 group) were treated with LY294002 in hypoxic environment. Cells in hypoxia without LY294002 were used as control (hypoxia group). The growth of U251 cells was detected by CCK-8 kit, the percentage of apoptotic cells was detected by FCM, the cell adhesion ability was detected by adhesion assay, the invasion ability of cells was detected by invasion assay, the expression of p-Akt protein was detected by Western blotting, RT- HIF-1α mRNA and protein expression. Results: LY294002 significantly inhibited the expression of p-Akt protein and HIF-1α mRNA and protein, which were significantly different from those in hypoxia group (P <0.05). LY294002 could inhibit the proliferation of U251 cells in hypoxic environment. The inhibitory effect of LY294002 was the most obvious on the 2nd to 6th day after the drug treatment (P <0.05). The percentage of apoptosis in hypoxia + LY294002 group was significantly higher than that in hypoxia group (P <0.01), while the cell adhesion and invasion ability was significantly lower than that in hypoxia group (P <0.01). CONCLUSION: LY294002 can inhibit the expression of HIF-1α mRNA and protein downstream of PI3K / Akt signaling pathway in hypoxia, inhibit the growth of human glioma U251 cells, promote apoptosis, and inhibit cell adhesion and invasion.