Advertisement

Journal of Molecular Medicine

, Volume 88, Issue 8, pp 817–827 | Cite as

Role of hypoxia-inducible transcription factor 1α for progression and chemosensitivity of murine hepatocellular carcinoma

  • Katjana Daskalow
  • Nadine Rohwer
  • Esther Raskopf
  • Evelyne Dupuy
  • Anja Kühl
  • Christoph Loddenkemper
  • Bertram Wiedenmann
  • Volker Schmitz
  • Thorsten CramerEmail author
Original article

Abstract

Hepatocellular carcinoma (HCC) is a hypervascularized tumor entity with association of arterial vessel density with poor prognosis. The hypoxia-inducible transcription factor HIF-1α represents a pivotal regulator of angiogenesis and is thought to determine the angiogenic nature of HCC. However, the precise role of HIF-1α during the pathogenesis of HCC remains elusive. We established a functional inactivation of HIF-1α in vitro and in vivo via RNAi and Cre/loxP-mediated recombination, respectively, to determine HIF-1α’s role for tumor growth and chemosensitivity in transgenic and orthotopic murine HCC models. HIF-1α-deficient HCC cells displayed significantly reduced anchorage-independent growth and enhanced sensitivity toward etoposide, while basic cellular proliferation was unaffected. Analysis of gross tumor growth failed to detect reduced growth of HIF-1α-deficient tumors in the orthotopic and the transgenic HCC model, respectively. In line with the in vitro data, treatment of HIF-1α-deficient tumors with etoposide resulted in greater antiproliferative efficacy when compared to wild-type mice. Taken together, our study does not support a pivotal role of HIF-1α for tumor growth and angiogenesis in two murine HCC models. However, our data point toward a significant function of HIF-1α in determining chemosensitivity of HCC and therefore warrant validation of HIF-1α-inhibitors as adjuvant therapeutic agents in clinical studies of human HCC.

Keywords

Hypoxia HIF-1α HCC Chemotherapy 

Notes

Acknowledgments

This work was supported by grants from the Deutsche Forschungsgemeinschaft (Cr 133/2-1, 133/2-2 and 133/2-3) and the Berliner Krebsgesellschaft (CRFF200804) to TC. NR was supported by the Deutsche Forschungsgemeinschaft (Graduiertenkolleg 276/4 - "Signalerkennung und -umsetzung").

Disclosure of potential conflict of interests

The authors declare no conflict of interests related to this study.

References

  1. 1.
    Llovet JM, Bruix J (2008) Molecular targeted therapies in hepatocellular carcinoma. Hepatology 4:1312–1327CrossRefGoogle Scholar
  2. 2.
    Roxburgh P, Evans TR (2008) Systemic therapy of hepatocellular carcinoma: are we making progress? Adv Ther 11:1089–1104CrossRefGoogle Scholar
  3. 3.
    Thomas MB (2008) Systemic therapy for hepatocellular carcinoma. Cancer J 2:123–127CrossRefGoogle Scholar
  4. 4.
    Llovet JM, Bruix J (2008) Novel advancements in the management of hepatocellular carcinoma in 2008. J Hepatol 48(Suppl 1):S20–S37CrossRefPubMedGoogle Scholar
  5. 5.
    Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A et al (2008) Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 4:378–390CrossRefGoogle Scholar
  6. 6.
    Harris AL (2002) Hypoxia—a key regulatory factor in tumour growth. Nat Rev Cancer 1:38–47CrossRefGoogle Scholar
  7. 7.
    Poellinger L, Johnson RS (2004) HIF-1 and hypoxic response: the plot thickens. Curr Opin Genet Dev 1:81–85CrossRefGoogle Scholar
  8. 8.
    Wang GL, Semenza GL (1993) General involvement of hypoxia-inducible factor 1 in transcriptional response to hypoxia. Proc Natl Acad Sci USA 9:4304–4308CrossRefGoogle Scholar
  9. 9.
    Wang GL, Jiang BH, Rue EA, Semenza GL (1995) Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci USA 12:5510–5514CrossRefGoogle Scholar
  10. 10.
    Semenza GL (2007) Hypoxia-inducible factor 1 (HIF-1) pathway. Sci STKE 407:cm8-Google Scholar
  11. 11.
    Weidemann A, Johnson RS (2008) Biology of HIF-1alpha. Cell Death Differ 4:621–627CrossRefGoogle Scholar
  12. 12.
    Semenza GL (2003) Targeting HIF-1 for cancer therapy. Nat Rev Cancer 10:721–732CrossRefGoogle Scholar
  13. 13.
    Daskalow K, Pfander D, Weichert W, Rohwer N, Thelen A, Neuhaus P, Jonas S, Wiedenmann B, Benckert C, Cramer T (2009) Distinct temporospatial expression patterns of glycolysis-related proteins in human hepatocellular carcinoma. Histochem Cell Biol 132(1):21–31CrossRefPubMedGoogle Scholar
  14. 14.
    Yasuda S, Arii S, Mori A, Isobe N, Yang W, Oe H, Fujimoto A, Yonenaga Y, Sakashita H, Imamura M (2004) Hexokinase II and VEGF expression in liver tumors: correlation with hypoxia-inducible factor 1 alpha and its significance. J Hepatol 1:117–123CrossRefGoogle Scholar
  15. 15.
    Kim KR, Moon HE, Kim KW (2002) Hypoxia-induced angiogenesis in human hepatocellular carcinoma. J Mol Med 11:703–714CrossRefGoogle Scholar
  16. 16.
    Rohwer N, Welzel M, Daskalow K, Pfander D, Wiedenmann B, Detjen K, Cramer T (2008) Hypoxia-inducible factor 1alpha mediates anoikis resistance via suppression of alpha5 integrin. Cancer Res 24:10113–10120CrossRefGoogle Scholar
  17. 17.
    Cramer T, Juttner S, Plath T, Mergler S, Seufferlein T, Wang TC, Merchant J, Hocker M (2008) Gastrin transactivates the chromogranin A gene through MEK-1/ERK- and PKC-dependent phosphorylation of Sp1 and CREB. Cell Signal 1:60–72CrossRefGoogle Scholar
  18. 18.
    Pastorekova S, Zavadova Z, Kostal M, Babusikova O, Zavada J (1992) A novel quasi-viral agent, MaTu, is a two-component system. Virology 2:620–626CrossRefGoogle Scholar
  19. 19.
    Cramer T, Yamanishi Y, Clausen BE, Forster I, Pawlinski R, Mackman N, Haase VH, Jaenisch R, Corr M, Nizet V et al (2003) HIF-1alpha is essential for myeloid cell-mediated inflammation. Cell 5:645–657CrossRefGoogle Scholar
  20. 20.
    Leurs C, Jansen M, Pollok KE, Heinkelein M, Schmidt M, Wissler M, Lindemann D, Von KC, Rethwilm A, Williams DA et al (2003) Comparison of three retroviral vector systems for transduction of nonobese diabetic/severe combined immunodeficiency mice repopulating human CD34+ cord blood cells. Hum Gene Ther 6:509–519CrossRefGoogle Scholar
  21. 21.
    Schmitz V, Tirado-Ledo L, Tiemann K, Raskopf E, Heinicke T, Ziske C, Gonzalez-Carmona MA, Rabe C, Wernert N, Prieto J et al (2004) Establishment of an orthotopic tumour model for hepatocellular carcinoma and non-invasive in vivo tumour imaging by high resolution ultrasound in mice. J Hepatol 5:787–791CrossRefGoogle Scholar
  22. 22.
    Dubois N, Bennoun M, Allemand I, Molina T, Grimber G, udet-Monsac M, Abelanet R, Briand P (1991) Time-course development of differentiated hepatocarcinoma and lung metastasis in transgenic mice. J Hepatol 2:227–239CrossRefGoogle Scholar
  23. 23.
    Ryan HE, Poloni M, McNulty W, Elson D, Gassmann M, Arbeit JM, Johnson RS (2000) Hypoxia-inducible factor-1alpha is a positive factor in solid tumor growth. Cancer Res 15:4010–4015Google Scholar
  24. 24.
    Postic C, Shiota M, Niswender KD, Jetton TL, Chen Y, Moates JM, Shelton KD, Lindner J, Cherrington AD, Magnuson MA (1999) Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic beta cell-specific gene knock-outs using Cre recombinase. J Biol Chem 1:305–315CrossRefGoogle Scholar
  25. 25.
    Rohwer N, Lobitz S, Daskalow K, Jons T, Vieth M, Schlag PM, Kemmner W, Wiedenmann B, Cramer T, Hocker M (2009) HIF-1alpha determines the metastatic potential of gastric cancer cells. Br J Cancer 5:772–781CrossRefGoogle Scholar
  26. 26.
    Takacova M, Barathova M, Hulikova A, Ohradanova A, Kopacek J, Parkkila S, Pastorek J, Pastorekova S, Zatovicova M (2007) Hypoxia-inducible expression of the mouse carbonic anhydrase IX demonstrated by new monoclonal antibodies. Int J Oncol 5:1103–1110Google Scholar
  27. 27.
    Nakashima Y, Nakashima O, Hsia CC, Kojiro M, Tabor E (1999) Vascularization of small hepatocellular carcinomas: correlation with differentiation. Liver 1:12–18CrossRefGoogle Scholar
  28. 28.
    Ueda K, Terada T, Nakanuma Y, Matsui O (1992) Vascular supply in adenomatous hyperplasia of the liver and hepatocellular carcinoma: a morphometric study. Hum Pathol 6:619–626CrossRefGoogle Scholar
  29. 29.
    Raskopf E, Vogt A, Sauerbruch T, Schmitz V (2008) siRNA targeting VEGF inhibits hepatocellular carcinoma growth and tumor angiogenesis in vivo. J Hepatol 6:977–984CrossRefGoogle Scholar
  30. 30.
    Frisch SM, Ruoslahti E (1997) Integrins and anoikis. Curr Opin Cell Biol 5:701–706CrossRefGoogle Scholar
  31. 31.
    Vogl TJ, Naguib NN, Nour-Eldin NE, Rao P, Emami AH, Zangos S, Nabil M, Abdelkader A (2009) Review on transarterial chemoembolization in hepatocellular carcinoma: palliative, combined, neoadjuvant, bridging, and symptomatic indications. Eur J Radiol 72(3):505–16CrossRefPubMedGoogle Scholar
  32. 32.
    Brown LM, Cowen RL, Debray C, Eustace A, Erler JT, Sheppard FCD, Parker CA, Stratford IJ, Williams KJ (2006) Reversing hypoxic cell chemoresistance in vitro using genetic and small molecule approaches targeting hypoxia inducible factor-1. Mol Pharmacol 2:411–418Google Scholar
  33. 33.
    Hao J, Song X, Song B, Liu Y, Wei L, Wang X, Yu J (2008) Effects of lentivirus-mediated HIF-1alpha knockdown on hypoxia-related cisplatin resistance and their dependence on p53 status in fibrosarcoma cells. Cancer Gene Ther 7:449–455CrossRefGoogle Scholar
  34. 34.
    Liu L, Ning X, Sun L, Zhang H, Shi Y, Guo C, Han S, Liu J, Sun S, Han Z et al (2008) Hypoxia-inducible factor-1 alpha contributes to hypoxia-induced chemoresistance in gastric cancer. Cancer Sci 1:121–128Google Scholar
  35. 35.
    Sasabe E, Zhou X, Li D, Oku N, Yamamoto T, Osaki T (2007) The involvement of hypoxia-inducible factor-1alpha in the susceptibility to gamma-rays and chemotherapeutic drugs of oral squamous cell carcinoma cells. Int J Cancer 2:268–277CrossRefGoogle Scholar
  36. 36.
    Sullivan R, Pare GC, Frederiksen LJ, Semenza GL, Graham CH (2008) Hypoxia-induced resistance to anticancer drugs is associated with decreased senescence and requires hypoxia-inducible factor-1 activity. Mol Cancer Ther 7:1961–1973CrossRefPubMedGoogle Scholar
  37. 37.
    Unruh A, Ressel A, Mohamed HG, Johnson RS, Nadrowitz R, Richter E, Katschinski DM, Wenger RH (2003) The hypoxia-inducible factor-1 alpha is a negative factor for tumor therapy. Oncogene 21:3213–3220CrossRefGoogle Scholar
  38. 38.
    Piret JP, Cosse JP, Ninane N, Raes M, Michiels C (2006) Hypoxia protects HepG2 cells against etoposide-induced apoptosis via a HIF-1-independent pathway. Exp Cell Res 15:2908–2920CrossRefGoogle Scholar
  39. 39.
    Yang ZF, Poon RT, To J, Ho DW, Fan ST (2004) The potential role of hypoxia inducible factor 1alpha in tumor progression after hypoxia and chemotherapy in hepatocellular carcinoma. Cancer Res 15:5496–5503CrossRefGoogle Scholar
  40. 40.
    Liu F, Wang P, Jiang X, Tan G, Qiao H, Jiang H, Krissansen GW, Sun X (2008) Antisense hypoxia-inducible factor 1alpha gene therapy enhances the therapeutic efficacy of doxorubicin to combat hepatocellular carcinoma. Cancer Sci 10:2055–2061Google Scholar
  41. 41.
    Tanaka H, Yamamoto M, Hashimoto N, Miyakoshi M, Tamakawa S, Yoshie M, Tokusashi Y, Yokoyama K, Yaginuma Y, Ogawa K (2006) Hypoxia-independent overexpression of hypoxia-inducible factor 1alpha as an early change in mouse hepatocarcinogenesis. Cancer Res 23:11263–11270CrossRefGoogle Scholar
  42. 42.
    Blouw B, Song H, Tihan T, Bosze J, Ferrara N, Gerber HP, Johnson RS, Bergers G (2003) The hypoxic response of tumors is dependent on their microenvironment. Cancer Cell 2:133–146CrossRefGoogle Scholar
  43. 43.
    Dupuy E, Hainaud P, Villemain A, Bodevin-Phedre E, Brouland JP, Briand P, Tobelem G (2003) Tumoral angiogenesis and tissue factor expression during hepatocellular carcinoma progression in a transgenic mouse model. J Hepatol 6:793–802CrossRefGoogle Scholar
  44. 44.
    Bangoura G, Liu ZS, Qian Q, Jiang CQ, Yang GF, Jing S (2007) Prognostic significance of HIF-2alpha/EPAS1 expression in hepatocellular carcinoma. World J Gastroenterol 23:3176–3182Google Scholar
  45. 45.
    Rankin EB, Rha J, Unger TL, Wu CH, Shutt HP, Johnson RS, Simon MC, Keith B, Haase VH (2008) Hypoxia-inducible factor-2 regulates vascular tumorigenesis in mice. Oncogene 40:5354–5358CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Katjana Daskalow
    • 1
  • Nadine Rohwer
    • 1
  • Esther Raskopf
    • 2
  • Evelyne Dupuy
    • 3
  • Anja Kühl
    • 4
  • Christoph Loddenkemper
    • 4
  • Bertram Wiedenmann
    • 1
  • Volker Schmitz
    • 2
  • Thorsten Cramer
    • 1
    • 5
    Email author
  1. 1.Medizinische Klinik mit Schwerpunkt Hepatologie und GastroenterologieCharité-Universitätsmedizin Berlin, Campus Virchow-KlinikumBerlinGermany
  2. 2.Medizinische Klinik und Poliklinik IUniversitätsklinikum BonnBonnGermany
  3. 3.Institut des Vaisseaux et du SangUniversité Paris 7-Denis Diderot, Hôpital LariboisièreParisFrance
  4. 4.Institut für PathologieCharité-Universitätsmedizin Berlin, Campus Benjamin FranklinBerlinGermany
  5. 5.Molekulares KrebsforschungszentrumCharité-Universitätsmedizin BerlinBerlinGermany

Personalised recommendations