Effects of siRNA-mediated HIF-1α gene silencing on angiogenesis in osteosarcoma

Objective: To explore angiogenesis in osteosarcoma under the condition of hypoxia-inducible factor (HIF)-1? gene silenced by small interference RNA (siRNA).Methods: The SaOS-2 osteosarcoma cells, transfected with the recombinant plasmid pSilencer2.1-HIF-1? or pSilencer2.1-SCR, were classified as HIF-1?/siRNA group or SCR/siRNA group, respectively. In which, vascular endothelial growth factor (VEGF) immunohistochemistry were performed. HIF-1? and VEGF protein contents were detected by western blot. Gene expressions of HIF-1? and VEGF were quantified by qPCR. Then the transfected SaOS-2 cells were inoculated in nude mice and transplantation tumor were checked via HE staining, VEGF and CD34 immunohistochemistry, and calculation of microvascular density (MVD).Results: In vitro, VEGF immunohistochemistry stains, HIF-1? and VEGF protein contents, and the relative expressions of HIF-1? mRNA and VEGF mRNA in HIF-1?/siRNA group were obviously reduced. In vivo, morphological observation illustrated that heteromorphism were not obvious in the cells of HIF-1?/siRNA group and vascular systems were sparse in its transplantation tumor tissue, and immunohistochemistry revealed that both VEGF and CD34 stains were significantly decreased in HIF-1?/siRNA group, and MVD in HIF-1?/siRNA group (7.3