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≤10-5 mol/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10-5 mol/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。
Objective To investigate the effects of oxidative stress on colon cancer cell migration,vascular epithelial growth factor expression and gap junction intercellular communication.Methods We 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.Results The 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,P0.05）,up-regulated the expression of VEGF gene（Z=3.938,P=0.000） and inhibited the intercellular communication（Z=3.938,P=0.000）.Conclusion Oxidative 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.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2011年第8期857-860,共4页
Cancer Research on Prevention and Treatment
基金
国家自然科学基金资助项目(30771126)
教育部留学回国人员基金资助项目(890)
江苏省自然科学基金资助项目(BK2006058)
关键词
氧化应激
大肠癌细胞
细胞迁移
血管内皮生长因子
细胞通信
Hydrogen peroxide
Colon cancer cell
Cell migration
Vascular epithelial growth factor
Gap junction intercellular communication
