缺氧对喉癌Hep-2细胞HIF-1α、GLUT-1、MMP-2表达的影响

目的 探讨缺氧对喉癌Hep-2细胞增殖、迁移和侵袭以及缺氧诱导因子-1α（HIF-1α）、葡萄糖转运蛋白-1（GLUT-1）和基质金属蛋白酶-2（MMP-2）表达的影响。方法 MTT法检测缺氧不同时间点细胞增殖状况；Transwell和划痕实验分别检测缺氧对喉癌Hep-2细胞侵袭和迁移能力的影响；Western blot和实时定量反转录聚合酶链反应（RT-PCR）检测缺氧后不同时间点HIF-1α、GLUT-1、MMP-2蛋白及mRNA表达的情况。结果 在缺氧条件下（1％O2、5％CO2和94％N2），Hep-2细胞的存活率随缺氧时间的延长逐渐升高，迁移和侵袭能力增强；随缺氧时间延长，Hep-2细胞HIF-1α蛋白表达水平明显增加，mRNA水平变化不明显；随缺氧时间延长，GLUT-1、MMP-2的mRNA水平和蛋白水平均明显增加。 结论 缺氧环境下，喉癌肿瘤细胞通过上调HIF-1α、GLUT-1、MMP-2的表达，强化其增殖、侵袭、转移的能力，促进喉癌的发展。

Effect of Hypoxia on Expression of HIF-1α, GLUT-1 and MMP-2 in Laryngeal Carcinoma Cell Line Hep-2

Objective To investigate cell proliferation, migration, invasion and the expression of HIF- 1α, GLUT-1, MMP-2 in laryngeal carcinoma cell line Hep-2 under hypoxia. Methods MTT assay was performed to investigate the proliferation of Hep-2 cells under hypoxic conditions; the invasion ability of Hep-2 cells was determined by Transwell assay; the migration ability of Hep-2 cells was determined by wound healing assay; the mRNA and protein expression of HIF-1α，MMP-2 and GLUT-1 were detected by Western blot and real-time PCR at various time points under hypoxia. Results Under the hypoxia(1％ O2, 5％CO2 and 94％N2), the growth activity of Hep-2 cell line was enhanced with the time prolonged, and the migration and invasion abilities of Hep-2 cell were enhanced. The protein expression of HIF-1α in the hypoxia was increased than that in the normoxia, however, HIF-1α mRNA levels had no significant difference between hypoxia and normoxia. The protein and mRNA expression of GLUT-1 and MMP-2 were increased in hypoxic conditions than those in the normoxia. Conclusion Hypoxic conditions may promote the protein expressions of HIF-1α, GLUT-1 and MMP-2 in laryngeal carcinoma cell line Hep-2; the overexpression of three proteins may have contributed to the proliferation, invasion and metastasis of laryngeal carcinoma cells.