This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Damert, A. Articles by Plate, K. H. Search for Related Content PubMed PubMed Citation Articles by Damert, A. Articles by Plate, K. H.

Up-Regulation of Vascular Endothelial Growth Factor Expression in a Rat Glioma Is Conferred by Two Distinct Hypoxia-driven Mechanisms¹

Max-Planck-Institut für Physiologische und Klinische Forschung, W. G. Kerckhoff-Institut, Abteilung Molekulare Zellbiologie, Parkstrasse 1, 61231 Bad Nauheim, Germany [A. D., G. B., W. R.]; Klinikum der Albert-Ludwigs-Universität Freiburg, Neurozentrum, Abteilung Neuropathologie, Breisacherstrasse 64, 79106 Freiburg, Germany [M. M., K. H. P.]; Department of Biochemistry and Biophysics, Hormone Research Institute, University of California at San Francisco, San Francisco, California 94143-0534 [M. Q. F., D. H.]

Up-regulation of vascular endothelial growth factor (VEGF) expression is a major event leading to neovascularization in malignant gliomas. Hypoxia is believed to be the crucial environmental stimulus for this up-regulation. To critically assess this hypothesis, we asked whether the mechanisms defined previously for hypoxia-induced VEGF expression in vitro are similarly involved and sufficient for up-regulation of VEGF gene expression in vivo, using a lacZ reporter gene under the control of VEGF regulatory sequences in an experimental glioma model. Inclusion of the binding site for hypoxia-inducible factor 1 (HIF 1) in the 5' regulatory sequences used in the hybrid gene produced weak ß-galactosidase staining in a special tumor cell subtype, the so-called perinecrotic palisading (PNP) cells that flank necrotic regions within the tumor. Deletion of the HIF 1 binding site abolished reporter gene expression in the PNP cells, indicating that transcriptional activation of VEGF expression in gliomas is mediated by HIF 1. Inclusion of 3' untranslated sequences from the VEGF gene in the reporter contructs resulted in an increased ß-galactosidase staining in the PNP cells, suggesting that mRNA stabilization also contributes to VEGF up-regulation in glioblastoma cells growing as solid tumors. Combination of the 5' flanking region including the HIF 1 site along with 3' untranslated sequences produced increased levels of ß-galactosidase expression in PNP cells. EF 5 immunostaining for regions of low oxygen partial pressure covered the same PNP cells that were stained for ß-galactosidase. Collectively, the data provide experimental evidence that VEGF gene expression is activated in a distinct tumor cell subpopulation, the perinecrotic palisading cells of gliomas, by two distinct hypoxia-driven regulatory mechanisms.

¹ This study was supported by the Deutsche Krebshilfe and the Bundesministerium für Bildung und Forschung. M. Q. F. was supported by NIH Grant 2PO1-DK41822.

² To whom requests for reprints should be addressed. Phone: 49-6032-705288; Fax: 49-6032-72259; E-mail: adamert@kerckhoff.mpg.de.

³ Present address: Department of Forensic Medicine and Pathology, Osaka City University, Medical School, Asahimachi 1-4-54, Abeno-ku, Osaka 545, Japan.

Received 3/21/97. Accepted 7/ 7/97.

This article has been cited by other articles:

				E. Karathanasis, S. Suryanarayanan, S. R. Balusu, K. McNeeley, I. Sechopoulos, A. Karellas, A. V. Annapragada, and R. V. Bellamkonda Imaging Nanoprobe for Prediction of Outcome of Nanoparticle Chemotherapy by Using Mammography Radiology, February 1, 2009; 250(2): 398 - 406. [Abstract] [Full Text] [PDF]

				Y. Zhang, N. Zhang, B. Dai, M. Liu, R. Sawaya, K. Xie, and S. Huang FoxM1B Transcriptionally Regulates Vascular Endothelial Growth Factor Expression and Promotes the Angiogenesis and Growth of Glioma Cells Cancer Res., November 1, 2008; 68(21): 8733 - 8742. [Abstract] [Full Text] [PDF]

				B. A. Bryan, T. E. Walshe, D. C. Mitchell, J. S. Havumaki, M. Saint-Geniez, A. S. Maharaj, A. E. Maldonado, and P. A. D'Amore Coordinated Vascular Endothelial Growth Factor Expression and Signaling During Skeletal Myogenic Differentiation Mol. Biol. Cell, March 1, 2008; 19(3): 994 - 1006. [Abstract] [Full Text] [PDF]

				E. Suswam, Y. Li, X. Zhang, G. Y. Gillespie, X. Li, J. J. Shacka, L. Lu, L. Zheng, and P. H. King Tristetraprolin Down-regulates Interleukin-8 and Vascular Endothelial Growth Factor in Malignant Glioma Cells Cancer Res., February 1, 2008; 68(3): 674 - 682. [Abstract] [Full Text] [PDF]

				J.M. Lupo, S. Cha, S.M. Chang, and S.J. Nelson Analysis of Metabolic Indices in Regions of Abnormal Perfusion in Patients with High-Grade Glioma AJNR Am. J. Neuroradiol., September 1, 2007; 28(8): 1455 - 1461. [Abstract] [Full Text] [PDF]

				J. D. Blais, C. L. Addison, R. Edge, T. Falls, H. Zhao, K. Wary, C. Koumenis, H. P. Harding, D. Ron, M. Holcik, et al. Perk-Dependent Translational Regulation Promotes Tumor Cell Adaptation and Angiogenesis in Response to Hypoxic Stress Mol. Cell. Biol., December 15, 2006; 26(24): 9517 - 9532. [Abstract] [Full Text] [PDF]

				D. B. Kirpotin, D. C. Drummond, Y. Shao, M. R. Shalaby, K. Hong, U. B. Nielsen, J. D. Marks, C. C. Benz, and J. W. Park Antibody Targeting of Long-Circulating Lipidic Nanoparticles Does Not Increase Tumor Localization but Does Increase Internalization in Animal Models. Cancer Res., July 1, 2006; 66(13): 6732 - 6740. [Abstract] [Full Text] [PDF]

				E. W. Newcomb, M. A. Ali, T. Schnee, L. Lan, Y. Lukyanov, M. Fowkes, D. C. Miller, and D. Zagzag Flavopiridol downregulates hypoxia-mediated hypoxia-inducible factor-1{alpha} expression in human glioma cells by a proteasome-independent pathway: Implications for in vivo therapy Neuro-oncol, July 1, 2005; 7(3): 225 - 235. [Abstract] [PDF]

				J. M. Lupo, S. Cha, S. M. Chang, and S. J. Nelson Dynamic Susceptibility-Weighted Perfusion Imaging of High-Grade Gliomas: Characterization of Spatial Heterogeneity AJNR Am. J. Neuroradiol., June 1, 2005; 26(6): 1446 - 1454. [Abstract] [Full Text] [PDF]

				T. Tanaka, T. Miyata, R. Inagi, T. Fujita, and M. Nangaku Hypoxia in Renal Disease with Proteinuria and/or Glomerular Hypertension Am. J. Pathol., December 1, 2004; 165(6): 1979 - 1992. [Abstract] [Full Text] [PDF]

				D. J. Brat, A. A. Castellano-Sanchez, S. B. Hunter, M. Pecot, C. Cohen, E. H. Hammond, S. N. Devi, B. Kaur, and E. G. Van Meir Pseudopalisades in Glioblastoma Are Hypoxic, Express Extracellular Matrix Proteases, and Are Formed by an Actively Migrating Cell Population Cancer Res., February 1, 2004; 64(3): 920 - 927. [Abstract] [Full Text] [PDF]

				V. Compernolle, K. Brusselmans, D. Franco, A. Moorman, M. Dewerchin, D. Collen, and P. Carmeliet Cardia bifida, defective heart development and abnormal neural crest migration in embryos lacking hypoxia-inducible factor-1{alpha} Cardiovasc Res, December 1, 2003; 60(3): 569 - 579. [Abstract] [Full Text] [PDF]

				A. Damert, L. Miquerol, M. Gertsenstein, W. Risau, and A. Nagy Insufficient VEGFA activity in yolk sac endoderm compromises haematopoietic and endothelial differentiation Development, March 6, 2003; 129(8): 1881 - 1892. [Abstract] [Full Text] [PDF]

				S. Matschurat, U. E. Knies, V. Person, L. Fink, B. Stoelcker, C. Ebenebe, H. A. Behrensdorf, J. Schaper, and M. Clauss Regulation of EMAP II by Hypoxia Am. J. Pathol., January 1, 2003; 162(1): 93 - 103. [Abstract] [Full Text] [PDF]

				T. Minami, D. J. Donovan, J. C. Tsai, R. D. Rosenberg, and W. C. Aird Differential regulation of the von Willebrand factor and Flt-1 promoters in the endothelium of hypoxanthine phosphoribosyltransferase-targeted mice Blood, December 1, 2002; 100(12): 4019 - 4025. [Abstract] [Full Text] [PDF]

				H. J. Schoch, S. Fischer, and H. H. Marti Hypoxia-induced vascular endothelial growth factor expression causes vascular leakage in the brain Brain, November 1, 2002; 125(11): 2549 - 2557. [Abstract] [Full Text] [PDF]

				M. Crowther, N. J. Brown, E. T. Bishop, and C. E. Lewis Microenvironmental influence on macrophage regulation of angiogenesis in wounds and malignant tumors J. Leukoc. Biol., October 1, 2001; 70(4): 478 - 490. [Abstract] [Full Text] [PDF]

				R. K. Bruick and S. L. McKnight Building better vasculature Genes & Dev., October 1, 2001; 15(19): 2497 - 2502. [Full Text] [PDF]

				K. Hattori, S. Dias, B. Heissig, N. R. Hackett, D. Lyden, M. Tateno, D. J. Hicklin, Z. Zhu, L. Witte, R. G. Crystal, et al. Vascular Endothelial Growth Factor and Angiopoietin-1 Stimulate Postnatal Hematopoiesis by Recruitment of Vasculogenic and Hematopoietic Stem Cells J. Exp. Med., May 7, 2001; 193(9): 1005 - 1014. [Abstract] [Full Text] [PDF]

				Q. Shi, X. Le, J. L. Abbruzzese, Z. Peng, C.-N. Qian, H. Tang, Q. Xiong, B. Wang, X.-C. Li, and K. Xie Constitutive Sp1 Activity Is Essential for Differential Constitutive Expression of Vascular Endothelial Growth Factor in Human Pancreatic Adenocarcinoma Cancer Res., May 1, 2001; 61(10): 4143 - 4154. [Abstract] [Full Text]

				S. P. Gunningham, M. J. Currie, C. Han, K. Turner, P. A. E. Scott, B. A. Robinson, A. L. Harris, and S. B. Fox Vascular Endothelial Growth Factor-B and Vascular Endothelial Growth Factor-C Expression in Renal Cell Carcinomas: Regulation by the von Hippel-Lindau Gene and Hypoxia Cancer Res., April 1, 2001; 61(7): 3206 - 3211. [Abstract] [Full Text]

				L. B. Nabors, G. Y. Gillespie, L. Harkins, and P. H. King HuR, a RNA Stability Factor, Is Expressed in Malignant Brain Tumors and Binds to Adenine- and Uridine-rich Elements within the 3' Untranslated Regions of Cytokine and Angiogenic Factor mRNAs Cancer Res., March 1, 2001; 61(5): 2154 - 2161. [Abstract] [Full Text]

				J. Grunstein, J. J. Masbad, R. Hickey, F. Giordano, and R. S. Johnson Isoforms of Vascular Endothelial Growth Factor Act in a Coordinate Fashion To Recruit and Expand Tumor Vasculature Mol. Cell. Biol., October 1, 2000; 20(19): 7282 - 7291. [Abstract] [Full Text]

				A. Maity, N. Pore, J. Lee, D. Solomon, and D. M. O’Rourke Epidermal Growth Factor Receptor Transcriptionally Up-Regulates Vascular Endothelial Growth Factor Expression in Human Glioblastoma Cells via a Pathway Involving Phosphatidylinositol 3'-Kinase and Distinct from That Induced by Hypoxia Cancer Res., October 1, 2000; 60(20): 5879 - 5886. [Abstract] [Full Text]

				M. Bongrazio, C. Baumann, A. Zakrzewicz, A. R Pries, and P. Gaehtgens Evidence for modulation of genes involved in vascular adaptation by prolonged exposure of endothelial cells to shear stress Cardiovasc Res, August 1, 2000; 47(2): 384 - 393. [Abstract] [Full Text] [PDF]

				L. Zhang, D. Yu, M. Hu, S. Xiong, A. Lang, L. M. Ellis, and R. E. Pollock Wild-Type p53 Suppresses Angiogenesis in Human Leiomyosarcoma and Synovial Sarcoma by Transcriptional Suppression of Vascular Endothelial Growth Factor Expression Cancer Res., July 1, 2000; 60(13): 3655 - 3661. [Abstract] [Full Text]

				N. Koshikawa, K. Takenaga, M. Tagawa, and S. Sakiyama Therapeutic Efficacy of the Suicide Gene Driven by the Promoter of Vascular Endothelial Growth Factor Gene against Hypoxic Tumor Cells Cancer Res., June 1, 2000; 60(11): 2936 - 2941. [Abstract] [Full Text] [PDF]

				M. Tenan, G. Fulci, M. Albertoni, A.-C. Diserens, M.-F. Hamou, M. El Atifi-Borel, J.-J. Feige, M. S. Pepper, and E. G. Van Meir Thrombospondin-1 Is Downregulated by Anoxia and Suppresses Tumorigenicity of Human Glioblastoma Cells J. Exp. Med., May 15, 2000; 191(10): 1789 - 1798. [Abstract] [Full Text] [PDF]

				P. H. King RNA-binding analyses of HuC and HuD with the VEGF and c-myc 3'-untranslated regions using a novel ELISA-based assay Nucleic Acids Res., April 1, 2000; 28(7): e20 - e20. [Abstract] [Full Text] [PDF]

				H. J. H. Marti, M. Bernaudin, A. Bellail, H. Schoch, M. Euler, E. Petit, and W. Risau Hypoxia-Induced Vascular Endothelial Growth Factor Expression Precedes Neovascularization after Cerebral Ischemia Am. J. Pathol., March 1, 2000; 156(3): 965 - 976. [Abstract] [Full Text] [PDF]

				M. Clauss and W. Schaper Vascular Endothelial Growth Factor : A Jack-of-All-Trades or a Nonspecific Stress Gene? Circ. Res., February 18, 2000; 86(3): 251 - 252. [Full Text] [PDF]

				S. R. Wedge, D. J. Ogilvie, M. Dukes, J. Kendrew, J. O. Curwen, L. F. Hennequin, A. P. Thomas, E. S. E. Stokes, B. Curry, G. H. P. Richmond, et al. ZD4190: An Orally Active Inhibitor of Vascular Endothelial Growth Factor Signaling with Broad-Spectrum Antitumor Efficacy Cancer Res., February 1, 2000; 60(4): 970 - 975. [Abstract] [Full Text]

				H. Kimura, A. Weisz, Y. Kurashima, K. Hashimoto, T. Ogura, F. D'Acquisto, R. Addeo, M. Makuuchi, and H. Esumi Hypoxia response element of the human vascular endothelial growth factor gene mediates transcriptional regulation by nitric oxide: control of hypoxia-inducible factor-1 activity by nitric oxide Blood, January 1, 2000; 95(1): 189 - 197. [Abstract] [Full Text] [PDF]

				S. A. Stacker, A. Vitali, C. Caesar, T. Domagala, L. C. Groenen, E. Nice, M. G. Achen, and A. F. Wilks A Mutant Form of Vascular Endothelial Growth Factor (VEGF) That Lacks VEGF Receptor-2 Activation Retains the Ability to Induce Vascular Permeability J. Biol. Chem., December 3, 1999; 274(49): 34884 - 34892. [Abstract] [Full Text] [PDF]

				L. Xu, K. Xie, N. Mukaida, K. Matsushima, and I. J. Fidler Hypoxia-induced Elevation in Interleukin-8 Expression by Human Ovarian Carcinoma Cells Cancer Res., November 1, 1999; 59(22): 5822 - 5829. [Abstract] [Full Text] [PDF]

				J. A. Dibbens, D. L. Miller, A. Damert, W. Risau, M. A. Vadas, and G. J. Goodall Hypoxic Regulation of Vascular Endothelial Growth Factor mRNA Stability Requires the Cooperation of Multiple RNA Elements Mol. Biol. Cell, April 1, 1999; 10(4): 907 - 919. [Abstract] [Full Text]

				G. Neufeld, T. Cohen, S. Gengrinovitch, and Z. Poltorak Vascular endothelial growth factor (VEGF) and its receptors FASEB J, January 1, 1999; 13(1): 9 - 22. [Abstract] [Full Text]

				H. H. Marti and W. Risau Systemic hypoxia changes the organ-specific distribution of vascular endothelial growth factor and its receptors PNAS, December 22, 1998; 95(26): 15809 - 15814. [Abstract] [Full Text] [PDF]

				M. Krieg, H. H. Marti, and K. H. Plate Coexpression of Erythropoietin and Vascular Endothelial Growth Factor in Nervous System Tumors Associated With von Hippel-Lindau Tumor Suppressor Gene Loss of Function Blood, November 1, 1998; 92(9): 3388 - 3393. [Abstract] [Full Text] [PDF]

				I. Flamme, M. Krieg, and K. H. Plate Up-Regulation of Vascular Endothelial Growth Factor in Stromal Cells of Hemangioblastomas Is Correlated with Up-Regulation of the Transcription Factor HRF/HIF-2{alpha} Am. J. Pathol., July 1, 1998; 153(1): 25 - 29. [Abstract] [Full Text] [PDF]

				D. Wang, D. B. Donner, and R. S. Warren Homeostatic Modulation of Cell Surface KDR and Flt1 Expression and Expression of the Vascular Endothelial Cell Growth Factor (VEGF) Receptor mRNAs by VEGF J. Biol. Chem., May 19, 2000; 275(21): 15905 - 15911. [Abstract] [Full Text] [PDF]