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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)-1α gene silenced by small interference RNA (siRNA). METHODS: The SaOS-2 osteosarcoma cells, transfected with the recombinant plasmid pSilencer2.1-HIF-1α or pSilencer2.1-SCR, were classified as HI...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Professional Medical Publications
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5432701/ https://www.ncbi.nlm.nih.gov/pubmed/28523034 http://dx.doi.org/10.12669/pjms.332.12587 |
Sumario: | OBJECTIVE: To explore angiogenesis in osteosarcoma under the condition of hypoxia-inducible factor (HIF)-1α gene silenced by small interference RNA (siRNA). METHODS: The SaOS-2 osteosarcoma cells, transfected with the recombinant plasmid pSilencer2.1-HIF-1α or pSilencer2.1-SCR, were classified as HIF-1α/siRNA group or SCR/siRNA group, respectively. In which, vascular endothelial growth factor (VEGF) immunohistochemistry were performed. HIF-1α and VEGF protein contents were detected by western blot. Gene expressions of HIF-1α 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-1α and VEGF protein contents, and the relative expressions of HIF-1α mRNA and VEGF mRNA in HIF-1α/siRNA group were obviously reduced. In vivo, morphological observation illustrated that heteromorphism were not obvious in the cells of HIF-1α/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-1α/siRNA group, and MVD in HIF-1α/siRNA group (7.3±1.1) were obviously less than that in SCR/siRNA group (17.2±3.2) (P<0.05). CONCLUSION: Angiogenesis in osteosarcoma can be inhibited by siRNA-mediated HIF-1α gene silencing, which is expected to provide a novel and attractive target of therapeutic strategies of osteosarcoma. |
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