c-myc通过提高结肠癌细胞LoVo的HIF-1α表达促进血管生成

Promoting Effect of c-myc on Angiogenesis through Increasing Expression of HIF-1α in LoVo Cell Line

目的研究癌基因c-myc对人结肠癌细胞LoVo中缺氧诱导因子-1α(HIF-1α)的影响及其对血管生成的作用。方法构建高表达c-myc的质粒pcDNA3.1-c-myc;将构建好的pcDNA3.1-c-myc质粒转染入LoVo细胞系中,通过Western blot检测c-myc在LoVo细胞中的表达情况。应用real-time PCR和Western blot检测常氧和乏氧状态下LoVo细胞系中HIF-1α、血管内皮生长因子(VEGF)的mRNA水平及HIF-1α的蛋白水平。应用条件培养液培养人脐静脉内皮细胞(HUVECs),观察其形成血管的能力。结果成功构建pcDNA3.1-c-myc质粒;常氧、乏氧状态下转染pcD-NA3.1-c-myc质粒后,LoVo细胞系中HIF-1α的mRNA水平无变化,VEGF的mRNA水平明显升高,HIF-1α蛋白水平明显增高。高表达c-myc的LoVo细胞上清液培养HUVECs细胞后能增强其成管能力。结论在LoVo细胞系中高表达c-myc,可以促进细胞HIF-1α和其下游的VEGF表达,从而促进血管生成。

Objective To investigate the influence of c-myc on HIF-1α in LoVo cell line and the effect on angiogenesis.Methods Plasmid pcDNA3.1-c-myc was constructed and transfected into LoVo cells.The transfection efficiency was tested by using Western blot.The mRNA expression level of HIF-1α and VEGF,and the protein expression level of HIF-1α were detected by using real-time PCR and Western blot under normoxia and hypoxia.Conditioned medium was applied to culture HUVECs and the ability of tube information was examined.Results The plasmid pcDNA3.1-c-myc was successfully constructed.Real-time PCR showed that the mRNA level of HIF-1α had no significant changes under normoxia and hypoxia.The protein expression level of HIF-1α was significantly increased,and the mRNA expression level of VEGF was obviously increased.The ability of tube formation by HUVECs was augmented through the overexpression of c-myc.Conclusion Overexpression of c-myc in LoVo cells can significantly increase the expression of HIF-1α and its downstream VEGF,and then promote angiogenesis.