Effect Of Hypoxia Inducible Factor 1α Antisense Oligonucleotides On The Expression Of Hypoxia Inducible Factor 1α And Vascular Endothelial Growth Factor In Osteosarcoma Cell Line MG-63

【Purpose】To investigate the effect of hypoxia inducible factor 1α(HIF-1α)antisense oligonucleotides(ASODN) on the expression of HIF-1αand vascular endothelial growth factor (VEGF)in osteosarcoma cell line MG-63 under hypoxia environment.【Methods】The ASODN and sense oligodeoxynucleotides(SODN) were designed on the basis of the HIF-1αcDNA sequence.The hypoxia culture model was established by a hypoxia incubator.The gene productions of HIF-1αand VEGF were observed at different hypoxia culture phase.The semi-quantitative reverse transcriotion PCR(RT-PCR) was used to test the mRNA expression of HIF-1αand VEGF.The protein level of HIF-1αand VEGF was observed by immunohistochemical staining and Western Blot analysis.【Results】Compared with the oxygen group,the mRNA level of HIF-1αwas not changed significantly,however,the protein expression of was increased remarkably with correspondence to the hypoxia time in the pure hypoxia group and the SODN hypoxia group;But the VEGF expression was up-regulated in mRNA and protein level.After treated by the ASODN,the mRNA and protein level of HIF-1αwas evidently decreased;the VEGF expression was also down-regulated in mRNA and protein level.Besides,the VEGF activity was markedly associated with the protein level of HIF-1α.【Conclusion】By hypoxia culture in vitro,the HIF-1αand VEGF genes could be up-regulated in osteosarcoma cell line MG-63.Furthermore,the expression of HIF-1αprotein can promote the VEGF activity. Nevertheless,the HIF-1αantisense oligonucleotides could inhibit the HIF-1αactivity ,which can reduce the VEGF expression in mRNA and protein level.