氧化应激对大肠癌细胞迁移、血管内皮生长因子表达及细胞间通信的影响

doi:10.3971/j.issn.1000-8578.2011.08.002

氧化应激对大肠癌细胞迁移、血管内皮生长因子表达及细胞间通信的影响

Effect of Oxidative Stress Cell Migration,Vascular Epithelial Growth Factor Expression and Gap Junction Intercellular Communication on Colon Cancer

摘要

摘要: 目的探讨氧化应激对大肠癌细胞迁移能力、血管内皮细胞生长因子表达及细胞间通信的影响。方法使用H2O2模拟大肠腔内氧化应激环境，通过不同浓度H₂O₂刺激人结肠癌细胞HCT-8，使癌细胞处于氧化应激状态，通过Transwell小室观察大肠癌细胞迁移能力；利用免疫细胞化学观察VEGF表达；利用划痕染料传输试验LY/DT法测定细胞间通信连接，最后利用Western blot进一步检测细胞间通信连接蛋白（Connexin 43）的表达。结果当H₂O₂浓度≤10-5mol/L时，不同程度地促进细胞的迁移，上调结肠癌细胞HCT-8 VEGF的表达，抑制细胞间通信；当H2O2浓度为10-5mol/L时, 明显促进细胞的迁移（t=2.247，P<0.05），上调结肠癌细胞HCT-8 VEGF的表达（Z=3.938，P=0.000），抑制细胞间通信(Z=3.860，P=0.000)。结论大肠癌细胞在过氧化物刺激的情况下，促进细胞迁移，诱导血管内皮生长，抑制细胞间通信，在大肠肿瘤的进展中可能具有促进作用。

Abstract: ObjectiveTo investigate the effects of oxidative stress on colon cancer cell migration, vascular epithelial growth factor expression and gap junction intercellular communication. MethodsWe stimulated HCT-8 human colon cancer cell with different concentration of hydrogen peroxide, through which mimicking the oxygen stress environment in the colon lumen was devedoped. We observed the colon cancer cell migration using Transwell Chamber assay, the expression of VEGF gene using immunocytochemistry, the gap junction intercellular communication using the scrape loading/dye transfer technique, and furthermore the expression of intercellular communication protein Connexin 43 using Western blot. ResultsThe oxygen stress environment induced by hydrogen peroxide at concentration ≤10-5mol/L shightly promoted and concentration of 10-5mol/L significantly promoted the colon cancer migration (t=2.247, P<0.05), up-regulated the expression of VEGF gene (Z=3.938, P=0.000) and inhibited the intercellular communication (Z=3.938, P=0.000). ConclusionOxidative stress mimicked by hydrogen peroxide might be involved in colorectal tumor development through promotion of the colon cancer cell migration, up-regulation of VEGF gene and inhibition of the intercellular communication.

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