Department of Otorhinolaryngology, Hebei Medical University, Shijiazhuang, Hebei 050017, P.R. China.
Mol Med Rep. 2012 Dec;6(6):1418-22. doi: 10.3892/mmr.2012.1075. Epub 2012 Sep 11.
The present study evaluated the regulation of glucose transporter protein-1 (Glut-1) and vascular endothelial growth factor (VEGF) by hypoxia inducible factor 1α (HIF-1α) under hypoxic conditions in Hep-2 human cells to explore the feasibility of these three genes as tumor markers. Hep-2 cells were cultured under hypoxic and normoxic conditions for 6, 12, 24, 36 and 48 h. The proliferation of Hep-2 cells was evaluated using an MTT assay. The protein and mRNA expression levels of HIF-1α, Glut-1 and VEGF were detected using the S-P immunocytochemical method, western blotting and reverse transcription polymerase chain reaction (RT-PCR). The results revealed that the expression levels of HIF-1α, Glut-1 and VEGF protein in Hep-2 cells were significantly elevated under hypoxic conditions compared with those under normoxic conditions over 36 h. Under hypoxic conditions, mRNA levels of HIF-1α were stable, while mRNA levels of Glut-1 and VEGF changed over time. In conclusion, Glut-1 and VEGF were upregulated by HIF-1α under hypoxic conditions in a time-dependent manner in Hep-2 cells and their co-expression serves as a tumor marker.
本研究在缺氧条件下评估了葡萄糖转运蛋白-1（Glut-1）和血管内皮生长因子（VEGF）受缺氧诱导因子 1α（HIF-1α）的调节，以探讨这三个基因作为肿瘤标志物的可行性。将 Hep-2 细胞在缺氧和常氧条件下分别培养 6、12、24、36 和 48 小时。使用 MTT 法评估 Hep-2 细胞的增殖。采用 S-P 免疫细胞化学法、western blot 法和逆转录聚合酶链反应（RT-PCR）检测 HIF-1α、Glut-1 和 VEGF 的蛋白和 mRNA 表达水平。结果表明，与常氧条件相比，缺氧条件下 Hep-2 细胞中 HIF-1α、Glut-1 和 VEGF 蛋白的表达水平在 36 小时以上显著升高。在缺氧条件下，HIF-1α 的 mRNA 水平稳定，而 Glut-1 和 VEGF 的 mRNA 水平随时间变化。综上所述，HIF-1α 可在 Hep-2 细胞中时间依赖性地上调 Glut-1 和 VEGF 的表达，其共同表达可作为肿瘤标志物。
