HIF-1α基因沉默对食管鳞癌细胞功能性基因的影响

Influence on functioning genes in esophageal squamous cancer cell after HIF-1α gene siliencing

目的观察食管鳞癌细胞中缺氧诱导因子-1α(HIF-1α)基因与血管内皮生长因子(VEGF)、血红素加氧酶(HO-1)、基质金属蛋白酶-2(MMP-2)基因之间的关系及其对细胞周期的影响。方法以HIF-1α基因沉默的Eca-109细胞、化学模拟缺氧细胞及未干预细胞为研究对象,通过Western blot检测VEGF、HO-1、MMP-2基因在三种细胞中的表达差异以了解其与HIF-1α基因的关系,并通过流式细胞术分析了解HIF-1α基因沉默后细胞周期的变化。结果Western blot检测发现,同空白及模拟缺氧组比较,HIF-1α基因沉默后HIF-1α蛋白表达明显下调,同时VEGF、HO-1、MMP-2的蛋白表达均出现了明显下降。流式细胞分析表明,HIF-1α基因沉默后的Eca-109细胞组同对照组相比G1、G0期细胞明显增加,G2、M期细胞变化不大,S期细胞比例明显减少。结论HIF-1α与上述三种基因存在明显相关性,进而可能影响着食管鳞癌的血管生成、侵袭、应激保护等生物学行为,HIF-1α沉默可将食管鳞癌细胞阻滞于G、G期。

Objective To determinate correlations of hypoxia-inducible factor-let with VEGF, HO-1, MMP-2 genes and cell cycle in esophageal squamous cancer cell. Methods Western blotting was used to examine the expression of HIF-1α, VEGF, HO-1 and MMP-2 in the cell clones with knock-down HIF-1α gene, control cells and the cells treated with 100μm cobalt chloride. Cell cycle was determined by flow cytometry. Results All of the expression levels of HIF-1α, VEGF, HO-1 and MMP-2 in cell groups with HIF-1α gene silencing were significantly lower than those in control groups （P 〈 0.05）. FCM showed that HIF-1α inhibition caused an arrest of the cell cycle at G1/G0 phase. Conclusions The VEGF, HO-1 and MMP-2 genes are positively correlated with HIF-1α in esophageal squamous cancer cell. The biological behaviour of angiogenesis, metaptosis, invasion and stress protection in esophageal squamous cancer cell could be suppressed by HIF- 1α gene silencing.