Home LiteratureArticle Details
PMID: 15899860 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Decreased growth of Vhl-/- fibrosarcomas is associated with elevated levels of cyclin kinase inhibitors p21 and p27.

Molecular and cellular biology ·Vol. 25 ·No. 11 ·2005-06-00 ·Pages 4565-78

Mack FA, Patel JH, Biju MP, Haase VH, Simon MC

Abstract

Inactivating mutations within the von Hippel-Lindau (VHL) tumor suppressor gene predispose patients to develop a variety of highly vascularized tumors. pVHL targets alpha subunits of the heterodimeric transcription factor hypoxia-inducible factor (HIF), a critical regulator of energy metabolism, angiogenesis, hematopoiesis, and oxygen (O(2)) delivery, for ubiquitin-mediated degradation in an O(2)-dependent manner. To investigate the role of Vhl in cellular proliferation and tumorigenesis, we utilized mouse embryonic fibroblasts (MEFs), a common tool for analyzing cell cycle regulation, and generated Vhl(-)(/)(-) MEF-derived fibrosarcomas. Surprisingly, growth of both Vhl(-)(/)(-) MEFs and fibrosarcomas was impaired, although tumor vascularity was increased. Decreased proliferation of Vhl(-)(/)(-) MEFs was correlated with an overexpression of cyclin kinase inhibitors (CKIs) p21 and p27. The transcription of p21 and p27 is inhibited by c-Myc; therefore, the induction of CKIs was attributed to the ability of HIF to antagonize c-Myc activity. Indeed, p21 mRNA levels were elevated under normoxia in Vhl(-)(/)(-) MEFs, while c-Myc transcriptional activity was markedly reduced. Gene silencing of HIF-1alpha by small interfering RNA reduced p21 and p27 protein and mRNA levels in Vhl(-)(/)(-) MEFs. The induction of p21 and p27, mediated by constitutive activation of the HIF pathway, provides a mechanism for the decreased proliferation rates of Vhl(-)(/)(-) MEFs and fibrosarcomas. These results demonstrate that a loss of pVHL can induce growth arrest in certain cells types, which suggests that additional genetic mutations are necessary for VHL-associated tumorigenesis.

MeSH Terms
Animals Apoptosis Cell Cycle/genetics,physiology Cell Cycle Proteins/genetics,metabolism Cell Proliferation Cyclin-Dependent Kinase Inhibitor p21 Cyclin-Dependent Kinase Inhibitor p27 Embryo, Mammalian/cytology Female Fibroblasts/metabolism Fibrosarcoma/genetics,metabolism,pathology Gene Silencing Hypoxia-Inducible Factor 1, alpha Subunit Mice Mice, Mutant Strains Mutation Proto-Oncogene Proteins c-myc/antagonists & inhibitors,genetics,metabolism RNA, Small Interfering/genetics,pharmacology Transcription Factors/genetics,metabolism Transcription, Genetic Transcriptional Activation Tumor Suppressor Proteins/genetics,metabolism,physiology Ubiquitin-Protein Ligases/genetics,physiology Von Hippel-Lindau Tumor Suppressor Protein
Chemicals
Cdkn1a protein, mouse Cdkn1b protein, mouse Cell Cycle Proteins Cyclin-Dependent Kinase Inhibitor p21 Hypoxia-Inducible Factor 1, alpha Subunit Myc protein, mouse Proto-Oncogene Proteins c-myc RNA, Small Interfering Transcription Factors Tumor Suppressor Proteins Cyclin-Dependent Kinase Inhibitor p27 Ubiquitin-Protein Ligases Von Hippel-Lindau Tumor Suppressor Protein VHL protein, mouse
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Mack Fiona A
University of Pennsylvania School of Medicine, Abramson Family Cancer Research Institute, Philadelphia, PA 19104, USA.
Patel Jagruti H
Biju Mangatt P
Haase Volker H
Simon M Celeste
References (42)
42 references, click to expand
  1. HIF activation identifies early lesions in VHL kidneys: evidence for site-specific tumor suppressor function in the nephron.
    Cancer Cell. 2002 Jun;1(5):459-68 PMID: 12124175
  2. The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis.
    Nature. 1999 May 20;399(6733):271-5 PMID: 10353251
  3. Regulation of microtubule stability by the von Hippel-Lindau tumour suppressor protein pVHL.
    Nat Cell Biol. 2003 Jan;5(1):64-70 PMID: 12510195
  4. Loss of pVHL is sufficient to cause HIF dysregulation in primary cells but does not promote tumor growth.
    Cancer Cell. 2003 Jan;3(1):75-88 PMID: 12559177
  5. Loss of von Hippel-Lindau protein causes cell density dependent deregulation of CyclinD1 expression through hypoxia-inducible factor.
    Oncogene. 2003 May 8;22(18):2728-38 PMID: 12743597
  6. Hypoxia-induced gene expression occurs solely through the action of hypoxia-inducible factor 1alpha (HIF-1alpha): role of cytoplasmic trapping of HIF-2alpha.
    Mol Cell Biol. 2003 Jul;23(14):4959-71 PMID: 12832481
  7. The hypoxic response of tumors is dependent on their microenvironment.
    Cancer Cell. 2003 Aug;4(2):133-46 PMID: 12957288
  8. Inhibition of hypoxia-inducible factor is sufficient for growth suppression of VHL-/- tumors.
    Mol Cancer Res. 2004 Feb;2(2):89-95 PMID: 14985465
  9. Gene array of VHL mutation and hypoxia shows novel hypoxia-induced genes and that cyclin D1 is a VHL target gene.
    Br J Cancer. 2004 Mar 22;90(6):1235-43 PMID: 15026807
  10. Inhibition of HIF2alpha is sufficient to suppress pVHL-defective tumor growth.
    PLoS Biol. 2003 Dec;1(3):E83 PMID: 14691554
  11. HIF-1alpha induces cell cycle arrest by functionally counteracting Myc.
    EMBO J. 2004 May 5;23(9):1949-56 PMID: 15071503
  12. Deletion of Vhlh in chondrocytes reduces cell proliferation and increases matrix deposition during growth plate development.
    Development. 2004 May;131(10):2497-508 PMID: 15128677
  13. Somatic mutations of the von Hippel-Lindau disease tumour suppressor gene in non-familial clear cell renal carcinoma.
    Hum Mol Genet. 1994 Dec;3(12):2169-73 PMID: 7881415
  14. Tumour suppression by the human von Hippel-Lindau gene product.
    Nat Med. 1995 Aug;1(8):822-6 PMID: 7585187
  15. Molecular cloning of the von Hippel-Lindau tumor suppressor gene and its role in renal carcinoma.
    Biochim Biophys Acta. 1996 Mar 18;1242(3):201-10 PMID: 8603073
  16. Cellular transformation and malignancy induced by ras require c-jun.
    Mol Cell Biol. 1996 Aug;16(8):4504-11 PMID: 8754851
  17. Negative regulation of hypoxia-inducible genes by the von Hippel-Lindau protein.
    Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10595-9 PMID: 8855223
  18. Investigation of c-myc and K-ras amplification in renal clear cell adenocarcinoma.
    Cancer Lett. 1997 Jan 1;111(1-2):127-31 PMID: 9022137
  19. Abnormal angiogenesis and responses to glucose and oxygen deprivation in mice lacking the protein ARNT.
    Nature. 1997 Mar 27;386(6623):403-7 PMID: 9121557
  20. Defective placental vasculogenesis causes embryonic lethality in VHL-deficient mice.
    Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):9102-7 PMID: 9256442
  21. von Hippel-Lindau disease.
    Medicine (Baltimore). 1997 Nov;76(6):381-91 PMID: 9413424
  22. The von Hippel-Lindau tumor suppressor gene is required for cell cycle exit upon serum withdrawal.
    Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):993-8 PMID: 9448273
  23. The role of ARNT2 in tumor angiogenesis and the neural response to hypoxia.
    Biochem Biophys Res Commun. 2000 Jun 24;273(1):231-8 PMID: 10873592
  24. Expression of the gene encoding the proapoptotic Nip3 protein is induced by hypoxia.
    Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):9082-7 PMID: 10922063
  25. Up-regulation of hypoxia-inducible factors HIF-1alpha and HIF-2alpha under normoxic conditions in renal carcinoma cells by von Hippel-Lindau tumor suppressor gene loss of function.
    Oncogene. 2000 Nov 16;19(48):5435-43 PMID: 11114720
  26. p21(Cip1) and p27(Kip1) regulate cell cycle reentry after hypoxic stress but are not necessary for hypoxia-induced arrest.
    Mol Cell Biol. 2001 Feb;21(4):1196-206 PMID: 11158306
  27. Vascular tumors in livers with targeted inactivation of the von Hippel-Lindau tumor suppressor.
    Proc Natl Acad Sci U S A. 2001 Feb 13;98(4):1583-8 PMID: 11171994
  28. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing.
    Science. 2001 Apr 20;292(5516):464-8 PMID: 11292862
  29. Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation.
    Science. 2001 Apr 20;292(5516):468-72 PMID: 11292861
  30. von Hippel-Lindau protein promotes the assembly of actin and vinculin and inhibits cell motility.
    Cancer Res. 2001 May 15;61(10):4184-9 PMID: 11358843
  31. Hypoxia inhibits G1/S transition through regulation of p27 expression.
    J Biol Chem. 2001 Mar 16;276(11):7919-26 PMID: 11112789
  32. HIF-1alpha binding to VHL is regulated by stimulus-sensitive proline hydroxylation.
    Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9630-5 PMID: 11504942
  33. HIF-1-dependent regulation of hypoxic induction of the cell death factors BNIP3 and NIX in human tumors.
    Cancer Res. 2001 Sep 15;61(18):6669-73 PMID: 11559532
  34. Hypoxia in cartilage: HIF-1alpha is essential for chondrocyte growth arrest and survival.
    Genes Dev. 2001 Nov 1;15(21):2865-76 PMID: 11691837
  35. The von Hippel-Lindau tumor suppressor protein mediates ubiquitination of activated atypical protein kinase C.
    J Biol Chem. 2001 Nov 23;276(47):43611-7 PMID: 11574546
  36. Phosphatidylinositol 3-kinase/Akt signaling is neither required for hypoxic stabilization of HIF-1 alpha nor sufficient for HIF-1-dependent target gene transcription.
    J Biol Chem. 2002 Apr 26;277(17):15162-70 PMID: 11859074
  37. VHL-mediated hypoxia regulation of cyclin D1 in renal carcinoma cells.
    Cancer Res. 2002 Jun 1;62(11):3014-9 PMID: 12036906
  38. Cell attachment to the extracellular matrix induces proteasomal degradation of p21(CIP1) via Cdc42/Rac1 signaling.
    Mol Cell Biol. 2002 Jul;22(13):4587-97 PMID: 12052868
  39. Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein.
    Cancer Cell. 2002 Apr;1(3):237-46 PMID: 12086860
  40. The contribution of VHL substrate binding and HIF1-alpha to the phenotype of VHL loss in renal cell carcinoma.
    Cancer Cell. 2002 Apr;1(3):247-55 PMID: 12086861
  41. The von Hippel-Lindau tumor suppressor protein is required for proper assembly of an extracellular fibronectin matrix.
    Mol Cell. 1998 Jun;1(7):959-68 PMID: 9651579
  42. Hypoxia-inducible factor 1alpha is essential for cell cycle arrest during hypoxia.
    Mol Cell Biol. 2003 Jan;23(1):359-69 PMID: 12482987
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.11.4565-4578.2005
Published
2005-06-00
Pages
4565-78
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1140627
Subset
IM
Grants
NCI NIH HHS · R01 CA100787 · United States
NCI NIH HHS · R01 CA100787-02 · United States
NHLBI NIH HHS · HL63310 · United States
NICHD NIH HHS · 1F31HD · United States
External Links
PubMed source DOI: 10.1128/MCB.25.11.4565-4578.2005 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2