1. ¹Microvascular and Molecular Neuro-Oncology Laboratory, New York University School of Medicine, New York, NY, USA
2. ²Department of Pathology, New York University School of Medicine, New York, NY, USA
3. ³Division of Neuropathology, New York University School of Medicine, New York, NY, USA
4. ⁴Department of Neurosurgery, New York University School of Medicine, New York, NY, USA
5. ⁵New York University Cancer Institute, New York University School of Medicine, New York, NY, USA

Correspondence: Dr D Zagzag, MD, PhD, Division of Neuropathology, Department of Pathology, New York University Medical Center, 550 First Avenue, New York, NY 10016, USA. E-mail: dz4@nyu.edu

Received 13 April 2006; Revised 9 May 2006; Accepted 9 June 2006; Published online 30 October 2006.

Abstract

Hypoxia and hypoxia-inducible factor-1 (HIF-1) play a critical role in glioblastoma multiforme (GBMs). CXCR4 is involved in angiogenesis and is upregulated by HIF-1. CXCR4 is a chemokine receptor for stromal cell-derived factor-1 (SDF-1), also known as CXCL12. We hypothesized that CXCR4 would be upregulated by hypoxia in GBMs. First, we investigated the expression of HIF-1 and CXCR4 in GBMs. CXCR4 was consistently found colocalized with HIF-1 expression in pseudopalisading glioma cells around areas of necrosis. In addition, angiogenic tumor vessels were strongly positive for CXCR4. Next, we tested the in vitro effect of hypoxia and vascular endothelial growth factor (VEGF) on the expression of CXCR4 in glioma cell lines and in human brain microvascular endothelial cells (HBMECs). Exposure to hypoxia induced significant expression of CXCR4 and HIF-1 in glioma cells, whereas treatment with exogenous VEGF increased CXCR4 expression in HBMECs. We also transfected U87MG glioma cells with an HIF-1 construct and observed that CXCR4 was upregulated in these cells even in normoxic conditions. We then used a lentivirus-mediated shRNA expression vector directed against HIF-1. When exposed to hypoxia, infected cells failed to show HIF-1 and CXCR4 upregulation. We performed migration assays under normoxic and hypoxic conditions in the presence or absence of AMD3100, a CXCR4 inhibitor. There was a significant increase in the migration of U87MG and LN308 glioma cells in hypoxic conditions, which was inhibited in the presence of AMD3100. These studies demonstrate the critical role played by hypoxia and CXCR4 in glioma cell migration. Based on these studies, we suggest that hypoxia regulates CXCR4 in GBMs at two levels. First, through HIF-1 in the pseudopalisading tumor cells themselves and, secondly, by the VEGF-stimulated angiogenic response in HBMECs. We believe this knowledge may lead to a potentially important two-pronged therapy against GBM progression using chemotherapy targeting CXCR4.