摘要
目的 研究缺氧诱导因子（HIF-1α）在mTOR特异性抑制剂RAD001联合DDP下调胃癌细胞株SGC7901/DDP VEGF表达中的作用,并探讨其相关作用机制.方法 体外培养胃癌SGC7901/DDP细胞株,分别给予RAD001、DDP及RAD001＋DDP干预48 h后,免疫细胞化学染色及Western-blot法检测HIF-1α、血管内皮生长因子（VEGF）蛋白的表达情况,RT-PCR法检测HIF-1α mRNA、VEGF mRNA的合成情况.结果 RAD001单独作用于SGC7901/DDP细胞48 h后,HIF-1α、VEGF蛋白的表达和HIF-1α 、VEGF mRNA的合成下降,且呈剂量依赖性.联合用药组比单独用药组作用增强,差异有统计学意义（P〈0.05）.VEGF和HIF-1α蛋白呈正相关（r=0.993,P〈0.01）,HIF-1α mRNA和VEGF mRNA呈正相关（r=0.994,P〈0.01）.结论 RAD001能够抑制细胞体外VEGF、HIF-1α的蛋白表达及mRNA的合成,RAD001联合DDP作用后,效果增强,其机制可能与mTOR/HIF-1α/VEGF信号通路有关.
Objective To investigate the effects of hypoxia - inducible factor 1α （ HIF - 1α） on downregulation of VEGF in SGC7901/DDP cells after treatment with the mTOR specific inhibitor RADO01 and DDP and its possible mecha- nism. Methods SGC7901/DDP cells were cultured in vitro and treated with DDP, RADO01 or RADO01 ＋ DDP for 48 hours. Then, immunocytochemistry and western blotting were used to detect the expression of VEGF and HIF - 1 α The synthesis of VEGF mRNA and HIF - 1 α mRNA were detected by RT - PCR. Results When the cells were treated with RADO01 alone for 48 hours, their expressions of HIF- 1α and VEGF at protein and mRNA levels were inhibited in a concentration -dependent manner. A combination of RAD001 and DDP showed stronger effects than either RADO01 or DDP alone （P 〈 0.05 ）. According to Pearson correlation analysis, the expression of VEGF and HIF - 1 α were positively correlated （r = 0. 993 ,P 〈 0.01 ）. The synthesis of VEGF mRNA and HIF- 1α mRNA were positively correlated （r = 0. 994, P 〈 0. 01 ）. Conclusion RADO01 alone could inhibit the expressions of HIF -1α and VEGF at both protein and mRNA levels. A combination of RADO01 and DDP showed stronger effects than RADO01 or DDP alone, which may be associated with the mTOR/HIF-1α/VEGF signaling pathway.
出处
《徐州医学院学报》
CAS
2014年第1期27-30,共4页
Acta Academiae Medicinae Xuzhou
基金
江苏省卫生厅课题（Z201016）
