Silencing of HIF-1alpha by RNA interference in human glioma cells in vitro and in vivo

Methods Mol Biol. 2009:487:283-301. doi: 10.1007/978-1-60327-547-7_14.

Abstract

Higher-grade gliomas are distinguished by increased vascular endothelial cell proliferation and peritumoral edema. These are thought to be instigated by vascular endothelial growth factor, which in turn is regulated by cellular oxygen tension. Hypoxia inducible factor-1alpha (HIF-1alpha) is a main responder to intracellular hypoxia and is overexpressed in many human cancers, including gliomas. Here we present methods for investigating the role of HIF-1alpha in glioma growth in vivo and in vitro using RNA interference in U251, U87, and U373 glioma cells.

MeSH terms

  • Animals
  • Blotting, Western
  • Brain Neoplasms / genetics
  • Brain Neoplasms / therapy*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation / drug effects
  • Genetic Therapy / methods
  • Glioma / genetics
  • Glioma / therapy*
  • Glucose Transporter Type 1 / metabolism
  • Humans
  • Hypoxia
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
  • Immunoenzyme Techniques
  • Mice
  • Mice, Nude
  • Polymerase Chain Reaction
  • RNA Interference*
  • RNA, Small Interfering / therapeutic use*
  • Transfection
  • Ubiquitin-Protein Ligases / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Glucose Transporter Type 1
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • RNA, Small Interfering
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • MIB1 ligase, human
  • Ubiquitin-Protein Ligases