中文摘要：

目的探讨磷脂酰肌醇-3-激酶（P13K）／哺乳动物雷帕霉素靶蛋白（mTOR）双靶点抑制剂PI-103与肿瘤坏死因子相关诱导凋亡配体（TRAIL）联合应用对喉鳞癌Hep-2细胞的作用。方法将喉癌Hep-2细胞分为7组，分别为P13K抑制剂LY294002组、mTOR抑制分子雷帕霉素组、P13K／mTOR双靶点抑制剂PI-103组、LY294002＋TRAIL组、雷帕霉素＋TRAIL组、PI-103＋TRAIL组和阴性对照组，以流式细胞仪检测各组Hep-2细胞周期及凋亡率。设立PI-103组、PI-103＋TRAIL组和阴性对照组，Transwell检测各组对Hep．2细胞迁移及侵袭能力影响，Western印迹法检测各组细胞中细胞凋亡及P13K／蛋白激酶B（AKT）／mTOR信号通路相关蛋白的表达水平。结果PI-103＋TRAIL组能够阻滞细胞周期于s期[G1：（1．80±0．30）％；G2：0．00]，相比其他各组能显著抑制细胞分裂，增强细胞凋亡[（66．78±2．93）％]（均P〈0．05）。PI-103＋TRAIL组细胞迁移数和侵袭数分别为（17．0±3．4）个和（18．4±5．4）个，PI．103组为（41．2±3．8）个和（41．6±4．7）个，联合用药组可显著降低细胞迁移及侵袭能力（均P〈0．05）。与对照组相比，PI．103处理组、PI-103＋TRAIL处理组半胱氨酸天冬氨酸蛋白酶（Caspase）-9、8、3的表达量逐渐升高，细胞周期蛋白（Cyclin）D1、CyclinE1、p-AKT、磷酸化真核翻译起始因子4E结合蛋白1（p-4E—BP1）的表达量逐渐降低（均P〈0．05）。结论PI-103与TRAIL联合应用于喉鳞癌Hep-2细胞能显著增强抗瘤效果，有望应用于喉鳞癌的治疗。

英文摘要：

Objective To investigate the mammalian target of rapamycin （mTOR） inhibitor, effects of a dual phosphoinosmde-3-kinase （PI3K）/ PI-103, cooperating with tumor necrosis factor-related apoptosis-indueing ligand （TRAIL） on the laryngeal squamous carcinoma Hep-2 cells. Methods Hep-2 cells were divided into 7 groups ： LY294002 group, Rapamyein group, PI-103 group, LY294002 ＋ TRAIL group, Rapamycin ＋ TRAIL group, PI-103 ＋ TRAIL group and control group. The cell cycle and apoptosis of Hep-2 cells were assessed by flow eytometry. For PI-103 group, PI-103 ＋ TRAIL group and control group, migration and invasion ability were measured by transwell migration and invasion assay respectively. The expression of relative proteins in apoptosis and PI3K/AKT/mTOR signal pathway was examined by Western blotting. Results Combination of PI-103 and TRAIL could make cell cycle arrest at S phase （ G1 ： 1.80% ＋ 0. 30% ;G2：0. 00）, inhibit cell proliferation, and enhance apoptosis （66. 78% ± 2. 93% ） （P 〈0. 05）. Combination of PI-103 and TRAIL could statistically decrease the migration and invasion number of Hep-2 cells （ 17.0 ± 3.4, 18.4 ± 5.4） than that of PI-103 group （41.2 ± 3.8, 41.6 ± 4. 7 ）. PI-103 could inhibit PI3K/AKT/mTOR signal pathway by decreasing the protein expression of p-AKT and p-4E-BP1. Comparing with the control group, the expression of eysteinyl aspartate specific proteinase （Caspase） 9, 8, 3 were increased while the expression of Cyelin D1, Cyclin E1, p-AKT, p-4E-BP1 were decreased in PI-103 and PI-103 ＋ TRAIL group （ P 〈 0. 05 ）. Conclusion Enhanced anti-tumor effects was observed by combination of PI-103 and TRAIL on laryngeal cancer cells in vitro and this combined administration might be a promising strategy for clinical treatment of laryngeal cancer.