VEGF expression in human breast carcinoma cell line induced by hypoxia and its mechanism / 重庆医科大学学报

ABSTRACT

Objective:To investigate the effect of hypoxia on vascular endothelial growth factor expression in breast carcinoma cell line(MD-MBA-435S) and its mechanism.Methods:Hypoxia culture model in vitro was established to test the expression of VEGF and HIF-1? and their correlation in MD-MBA-435S cell line.After cultured in hypoxia circumstance for a given period of time(4 hours,8 hours,12 hours,16 hours,20 hours),VEGF expression was tested in mRNA level and protein level by RT-PCR and Western blot analysis,HIF-1 ? expression tested in protein level.Cell viability in twenty hours was tested by trypan blue exclusion assay.Results:Cell survival was affected a little within 20 hours under hypoxia.Two isoforms of VEGFmRNA,VEGF-121 and VEGF165,were identified by RT-PCR,and their expressions increased after treatment of hypoxia for 4 hours.VEGF and HIF-1 ? protein expressions increased equally with VEGFmRNA confirmed by immunocytochemistry and Western blot and its positive correlation was found by Pearson product-moment correlation analysis.Conclusion:VEGF gene and protein can be induced remarkably by hypoxia in breast carcinoma cell line,which is regulated mostly by transcription factor HIF-1 ?.This mechanism may be involved in the malignant proliferation and transformation of breast neoplasm.