Effect of Hypoxia on Expression of HIF-1α, GLUT-1 and MMP-2 in Laryngeal Carcinoma Cell Line Hep-2
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摘要: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。
Abstract:ObjectiveTo investigate cell proliferation, migration, invasion and the expression of HIF-1α, GLUT-1, MMP-2 in laryngeal carcinoma cell line Hep-2 under hypoxia.
MethodsMTT 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.
ResultsUnder 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.
ConclusionHypoxic 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.
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图 1 MTT法检测缺氧对Hep-2细胞增殖的影响（OD值）
Figure 1 Effect of hypoxia on proliferation of Hep-2 cells detected by MTT (OD value)
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图 2 缺氧对喉癌Hep-2细胞侵袭能力的影响
Figure 2 Effect of hypoxia on invasive ability of laryngeal carcinoma cells Hep-2 detected by Transwell chamber assay
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图 3 缺氧对喉癌Hep-2细胞迁移能力的影响
Figure 3 Effect of hypoxia on migration ability of laryngeal carcinoma Hep-2 cells detected by wound healing assay
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图 4 缺氧不同时间点HIF-1α、GLUT-l、MMP-2 mRNA在Hep-2细胞中的表达
Figure 4 mRNA expression of HIF-1α, GLUT-l and MMP-2 in Hep-2 cells at various time points under hypoxia
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