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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 investigate the expression of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF), heme oxygenase (HO-1) and matrix metalloproteinase-2 The relationship between the cell cycle and its impact. METHODS: The expression of VEGF, HO-1 and MMP-2 in three types of cells was detected by Western blot in order to understand the difference between the expression of HIF-1α gene silencing Eca-109 cells, chemically simulated hypoxic cells and non-interfering cells Its relationship with HIF-1αgene, and through flow cytometry analysis of HIF-1α gene silencing cell cycle changes. Results Western blot showed that HIF-1α protein expression was significantly down-regulated after HIF-1α gene silencing compared with blank and hypoxia groups, and the protein expressions of VEGF, HO-1 and MMP-2 were significantly decreased. Flow cytometry analysis showed that compared with the control group, the number of cells in G1 phase and G0 phase in Eca-109 cells after silencing HIF-1α gene was significantly increased, while the cells in G2 phase and M phase did not change much, and the proportion of S phase cells was significantly decreased. Conclusion HIF-1α is significantly correlated with the above three genes, which may affect the biological behavior of esophageal squamous cell carcinoma, such as angiogenesis, invasion and stress protection. HIF-1α silencing can block esophageal squamous cell carcinoma cells in G, G period.