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. 1998 Jun 1;17(11):3005–3015. doi: 10.1093/emboj/17.11.3005

HIF-1 alpha is required for solid tumor formation and embryonic vascularization.

H E Ryan 1, J Lo 1, R S Johnson 1
PMCID: PMC1170640  PMID: 9606183

Abstract

The transcriptional response to lowered oxygen levels is mediated by the hypoxia-inducible transcription factor (HIF-1), a heterodimer consisting of the constitutively expressed aryl hydrocarbon receptor nuclear translocator (ARNT) and the hypoxic response factor HIF-1alpha. To study the role of the transcriptional hypoxic response in vivo we have targeted the murine HIF-1alpha gene. Loss of HIF-1alpha in embryonic stem (ES) cells dramatically retards solid tumor growth; this is correlated with a reduced capacity to release the angiogenic factor vascular endothelial growth factor (VEGF) during hypoxia. HIF-1alpha null mutant embryos exhibit clear morphological differences by embryonic day (E) 8.0, and by E8.5 there is a complete lack of cephalic vascularization, a reduction in the number of somites, abnormal neural fold formation and a greatly increased degree of hypoxia (measured by the nitroimidazole EF5). These data demonstrate the essential role of HIF-1alpha in controlling both embryonic and tumorigenic responses to variations in microenvironmental oxygenation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akazawa S., Unterman T., Metzger B. E. Glucose metabolism in separated embryos and investing membranes during organogenesis in the rat. Metabolism. 1994 Jul;43(7):830–835. doi: 10.1016/0026-0495(94)90262-3. [DOI] [PubMed] [Google Scholar]
  2. Banai S., Shweiki D., Pinson A., Chandra M., Lazarovici G., Keshet E. Upregulation of vascular endothelial growth factor expression induced by myocardial ischaemia: implications for coronary angiogenesis. Cardiovasc Res. 1994 Aug;28(8):1176–1179. doi: 10.1093/cvr/28.8.1176. [DOI] [PubMed] [Google Scholar]
  3. Brown J. M., Giaccia A. J. Tumour hypoxia: the picture has changed in the 1990s. Int J Radiat Biol. 1994 Jan;65(1):95–102. doi: 10.1080/09553009414550131. [DOI] [PubMed] [Google Scholar]
  4. Bunn H. F., Poyton R. O. Oxygen sensing and molecular adaptation to hypoxia. Physiol Rev. 1996 Jul;76(3):839–885. doi: 10.1152/physrev.1996.76.3.839. [DOI] [PubMed] [Google Scholar]
  5. Carmeliet P., Ferreira V., Breier G., Pollefeyt S., Kieckens L., Gertsenstein M., Fahrig M., Vandenhoeck A., Harpal K., Eberhardt C. Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature. 1996 Apr 4;380(6573):435–439. doi: 10.1038/380435a0. [DOI] [PubMed] [Google Scholar]
  6. Dachs G. U., Patterson A. V., Firth J. D., Ratcliffe P. J., Townsend K. M., Stratford I. J., Harris A. L. Targeting gene expression to hypoxic tumor cells. Nat Med. 1997 May;3(5):515–520. doi: 10.1038/nm0597-515. [DOI] [PubMed] [Google Scholar]
  7. Damert A., Machein M., Breier G., Fujita M. Q., Hanahan D., Risau W., Plate K. H. Up-regulation of vascular endothelial growth factor expression in a rat glioma is conferred by two distinct hypoxia-driven mechanisms. Cancer Res. 1997 Sep 1;57(17):3860–3864. [PubMed] [Google Scholar]
  8. Ema M., Taya S., Yokotani N., Sogawa K., Matsuda Y., Fujii-Kuriyama Y. A novel bHLH-PAS factor with close sequence similarity to hypoxia-inducible factor 1alpha regulates the VEGF expression and is potentially involved in lung and vascular development. Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4273–4278. doi: 10.1073/pnas.94.9.4273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fernandez-Salguero P., Pineau T., Hilbert D. M., McPhail T., Lee S. S., Kimura S., Nebert D. W., Rudikoff S., Ward J. M., Gonzalez F. J. Immune system impairment and hepatic fibrosis in mice lacking the dioxin-binding Ah receptor. Science. 1995 May 5;268(5211):722–726. doi: 10.1126/science.7732381. [DOI] [PubMed] [Google Scholar]
  10. Ferrara N., Carver-Moore K., Chen H., Dowd M., Lu L., O'Shea K. S., Powell-Braxton L., Hillan K. J., Moore M. W. Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature. 1996 Apr 4;380(6573):439–442. doi: 10.1038/380439a0. [DOI] [PubMed] [Google Scholar]
  11. Finkenzeller G., Technau A., Marmé D. Hypoxia-induced transcription of the vascular endothelial growth factor gene is independent of functional AP-1 transcription factor. Biochem Biophys Res Commun. 1995 Mar 8;208(1):432–439. doi: 10.1006/bbrc.1995.1356. [DOI] [PubMed] [Google Scholar]
  12. Forsythe J. A., Jiang B. H., Iyer N. V., Agani F., Leung S. W., Koos R. D., Semenza G. L. Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol. 1996 Sep;16(9):4604–4613. doi: 10.1128/mcb.16.9.4604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gassmann M., Kvietikova I., Rolfs A., Wenger R. H. Oxygen- and dioxin-regulated gene expression in mouse hepatoma cells. Kidney Int. 1997 Feb;51(2):567–574. doi: 10.1038/ki.1997.81. [DOI] [PubMed] [Google Scholar]
  14. Graeber T. G., Osmanian C., Jacks T., Housman D. E., Koch C. J., Lowe S. W., Giaccia A. J. Hypoxia-mediated selection of cells with diminished apoptotic potential in solid tumours. Nature. 1996 Jan 4;379(6560):88–91. doi: 10.1038/379088a0. [DOI] [PubMed] [Google Scholar]
  15. Hirose K., Morita M., Ema M., Mimura J., Hamada H., Fujii H., Saijo Y., Gotoh O., Sogawa K., Fujii-Kuriyama Y. cDNA cloning and tissue-specific expression of a novel basic helix-loop-helix/PAS factor (Arnt2) with close sequence similarity to the aryl hydrocarbon receptor nuclear translocator (Arnt). Mol Cell Biol. 1996 Apr;16(4):1706–1713. doi: 10.1128/mcb.16.4.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hogenesch J. B., Chan W. K., Jackiw V. H., Brown R. C., Gu Y. Z., Pray-Grant M., Perdew G. H., Bradfield C. A. Characterization of a subset of the basic-helix-loop-helix-PAS superfamily that interacts with components of the dioxin signaling pathway. J Biol Chem. 1997 Mar 28;272(13):8581–8593. doi: 10.1074/jbc.272.13.8581. [DOI] [PubMed] [Google Scholar]
  17. Hunter E. S., 3rd, Tugman J. A. Inhibitors of glycolytic metabolism affect neurulation-staged mouse conceptuses in vitro. Teratology. 1995 Dec;52(6):317–323. doi: 10.1002/tera.1420520602. [DOI] [PubMed] [Google Scholar]
  18. Ikeda E., Achen M. G., Breier G., Risau W. Hypoxia-induced transcriptional activation and increased mRNA stability of vascular endothelial growth factor in C6 glioma cells. J Biol Chem. 1995 Aug 25;270(34):19761–19766. doi: 10.1074/jbc.270.34.19761. [DOI] [PubMed] [Google Scholar]
  19. Ladoux A., Frelin C. Hypoxia is a strong inducer of vascular endothelial growth factor mRNA expression in the heart. Biochem Biophys Res Commun. 1993 Sep 15;195(2):1005–1010. doi: 10.1006/bbrc.1993.2144. [DOI] [PubMed] [Google Scholar]
  20. Lee J., Siemann D. W., Koch C. J., Lord E. M. Direct relationship between radiobiological hypoxia in tumors and monoclonal antibody detection of EF5 cellular adducts. Int J Cancer. 1996 Jul 29;67(3):372–378. doi: 10.1002/(SICI)1097-0215(19960729)67:3<372::AID-IJC11>3.0.CO;2-A. [DOI] [PubMed] [Google Scholar]
  21. Levy A. P., Levy N. S., Goldberg M. A. Post-transcriptional regulation of vascular endothelial growth factor by hypoxia. J Biol Chem. 1996 Feb 2;271(5):2746–2753. doi: 10.1074/jbc.271.5.2746. [DOI] [PubMed] [Google Scholar]
  22. Levy A. P., Levy N. S., Wegner S., Goldberg M. A. Transcriptional regulation of the rat vascular endothelial growth factor gene by hypoxia. J Biol Chem. 1995 Jun 2;270(22):13333–13340. doi: 10.1074/jbc.270.22.13333. [DOI] [PubMed] [Google Scholar]
  23. Liu Y., Cox S. R., Morita T., Kourembanas S. Hypoxia regulates vascular endothelial growth factor gene expression in endothelial cells. Identification of a 5' enhancer. Circ Res. 1995 Sep;77(3):638–643. doi: 10.1161/01.res.77.3.638. [DOI] [PubMed] [Google Scholar]
  24. Lord E. M., Harwell L., Koch C. J. Detection of hypoxic cells by monoclonal antibody recognizing 2-nitroimidazole adducts. Cancer Res. 1993 Dec 1;53(23):5721–5726. [PubMed] [Google Scholar]
  25. Maltepe E., Schmidt J. V., Baunoch D., Bradfield C. A., Simon M. C. Abnormal angiogenesis and responses to glucose and oxygen deprivation in mice lacking the protein ARNT. Nature. 1997 Mar 27;386(6623):403–407. doi: 10.1038/386403a0. [DOI] [PubMed] [Google Scholar]
  26. Matsumoto K., Akazawa S., Ishibashi M., Trocino R. A., Matsuo H., Yamasaki H., Yamaguchi Y., Nagamatsu S., Nagataki S. Abundant expression of GLUT1 and GLUT3 in rat embryo during the early organogenesis period. Biochem Biophys Res Commun. 1995 Apr 6;209(1):95–102. doi: 10.1006/bbrc.1995.1475. [DOI] [PubMed] [Google Scholar]
  27. Mombaerts P., Iacomini J., Johnson R. S., Herrup K., Tonegawa S., Papaioannou V. E. RAG-1-deficient mice have no mature B and T lymphocytes. Cell. 1992 Mar 6;68(5):869–877. doi: 10.1016/0092-8674(92)90030-g. [DOI] [PubMed] [Google Scholar]
  28. Mortensen R. M., Conner D. A., Chao S., Geisterfer-Lowrance A. A., Seidman J. G. Production of homozygous mutant ES cells with a single targeting construct. Mol Cell Biol. 1992 May;12(5):2391–2395. doi: 10.1128/mcb.12.5.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pe'er J., Shweiki D., Itin A., Hemo I., Gnessin H., Keshet E. Hypoxia-induced expression of vascular endothelial growth factor by retinal cells is a common factor in neovascularizing ocular diseases. Lab Invest. 1995 Jun;72(6):638–645. [PubMed] [Google Scholar]
  30. Pötgens A. J., Lubsen N. H., van Altena M. C., Schoenmakers J. G., Ruiter D. J., de Waal R. M. Vascular permeability factor expression influences tumor angiogenesis in human melanoma lines xenografted to nude mice. Am J Pathol. 1995 Jan;146(1):197–209. [PMC free article] [PubMed] [Google Scholar]
  31. Rowlands J. C., Gustafsson J. A. Aryl hydrocarbon receptor-mediated signal transduction. Crit Rev Toxicol. 1997 Mar;27(2):109–134. doi: 10.3109/10408449709021615. [DOI] [PubMed] [Google Scholar]
  32. Schlaeger T. M., Qin Y., Fujiwara Y., Magram J., Sato T. N. Vascular endothelial cell lineage-specific promoter in transgenic mice. Development. 1995 Apr;121(4):1089–1098. doi: 10.1242/dev.121.4.1089. [DOI] [PubMed] [Google Scholar]
  33. Schmidt J. V., Su G. H., Reddy J. K., Simon M. C., Bradfield C. A. Characterization of a murine Ahr null allele: involvement of the Ah receptor in hepatic growth and development. Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6731–6736. doi: 10.1073/pnas.93.13.6731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Semenza G. L., Nejfelt M. K., Chi S. M., Antonarakis S. E. Hypoxia-inducible nuclear factors bind to an enhancer element located 3' to the human erythropoietin gene. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5680–5684. doi: 10.1073/pnas.88.13.5680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Semenza G. L., Roth P. H., Fang H. M., Wang G. L. Transcriptional regulation of genes encoding glycolytic enzymes by hypoxia-inducible factor 1. J Biol Chem. 1994 Sep 23;269(38):23757–23763. [PubMed] [Google Scholar]
  36. Semenza G. L., Wang G. L. A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcriptional activation. Mol Cell Biol. 1992 Dec;12(12):5447–5454. doi: 10.1128/mcb.12.12.5447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shepard T. H., Tanimura T., Park H. W. Glucose absorption and utilization by rat embryos. Int J Dev Biol. 1997 Apr;41(2):307–314. [PubMed] [Google Scholar]
  38. Shima D. T., Deutsch U., D'Amore P. A. Hypoxic induction of vascular endothelial growth factor (VEGF) in human epithelial cells is mediated by increases in mRNA stability. FEBS Lett. 1995 Aug 21;370(3):203–208. doi: 10.1016/0014-5793(95)00831-s. [DOI] [PubMed] [Google Scholar]
  39. Shweiki D., Itin A., Soffer D., Keshet E. Vascular endothelial growth factor induced by hypoxia may mediate hypoxia-initiated angiogenesis. Nature. 1992 Oct 29;359(6398):843–845. doi: 10.1038/359843a0. [DOI] [PubMed] [Google Scholar]
  40. Stone J., Itin A., Alon T., Pe'er J., Gnessin H., Chan-Ling T., Keshet E. Development of retinal vasculature is mediated by hypoxia-induced vascular endothelial growth factor (VEGF) expression by neuroglia. J Neurosci. 1995 Jul;15(7 Pt 1):4738–4747. doi: 10.1523/JNEUROSCI.15-07-04738.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Swanson H. I., Bradfield C. A. The AH-receptor: genetics, structure and function. Pharmacogenetics. 1993 Oct;3(5):213–230. doi: 10.1097/00008571-199310000-00001. [DOI] [PubMed] [Google Scholar]
  42. Tian H., McKnight S. L., Russell D. W. Endothelial PAS domain protein 1 (EPAS1), a transcription factor selectively expressed in endothelial cells. Genes Dev. 1997 Jan 1;11(1):72–82. doi: 10.1101/gad.11.1.72. [DOI] [PubMed] [Google Scholar]
  43. Wang G. L., Jiang B. H., Rue E. A., Semenza G. L. Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5510–5514. doi: 10.1073/pnas.92.12.5510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wang G. L., Semenza G. L. Characterization of hypoxia-inducible factor 1 and regulation of DNA binding activity by hypoxia. J Biol Chem. 1993 Oct 15;268(29):21513–21518. [PubMed] [Google Scholar]
  45. Wang G. L., Semenza G. L. General involvement of hypoxia-inducible factor 1 in transcriptional response to hypoxia. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4304–4308. doi: 10.1073/pnas.90.9.4304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wang G. L., Semenza G. L. Purification and characterization of hypoxia-inducible factor 1. J Biol Chem. 1995 Jan 20;270(3):1230–1237. doi: 10.1074/jbc.270.3.1230. [DOI] [PubMed] [Google Scholar]

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