Medical Oncology

, Volume 25, Issue 1, pp 88–92 | Cite as

Inhibitory effect of Hypoxia inducible factor-1 antisense oligonucleotide on growth of human hepatocellular carcinoma cells

  • Chen WeiXing
  • Hu Tiantian
  • Ni Qun
  • Yu Chaohui
  • Xu Ping
Original Paper



To observe the inhibitory effect of Hypoxia inducible factor-1 antisense oligonuclecotide on growth of human hepatocellular carcinoma cells and gene expression of HIF-1, in order to seek a new gene therapy for hepatocellular carcinoma.


Six Hypoxia inducible factor-1 antisense oligonuclecotides with various concentrations (0.2 μmol/l, 0.4 μmol/l, and 0.8 μmol/l) were transformed into HepG2 cells by lipofectamine reagent. 72 h after transfection, MTS assay was used to detect cellular proliferation. In addition, Hypoxia inducible factor-1 antisense oligonuclecotide2 with various concentrations (0.2, 0.4, 0.8, and 1.0 μmol/l) were transformed into HepG2 cells. About 48 h after transfection, reverse transcription-polymerase chain reaction assay and Western Blot assay were employed to detect the expression of Hypoxia inducible factor-1 gene and the synthesis of Hypoxia inducible factor-1 protein respectively.


HepG2 cell growth was inhibited by 6 Hypoxia inducible factor-1 antisense oligonuclecotides at various concentrations. Among them, Hypoxia inducible factor-1 antisense oligonuclecotide2 showed the most effective inhibition ability (P < 0.01), the inhibitory rate was 89.66% at the concentration of 1.0 μmol/l. About 48 h after transfection, Hypoxia inducible factor-1 mRNA expression was downregulated and Hypoxia inducible factor-1 protein synthesis was decreased by antisense oligonuclecotide2.


The hepatocellular carcinoma cell proliferation was inhibited by Hypoxia inducible factor-1 antisense oligonuclecotide. Moreover, the gene expression and protein synthesis of Hypoxia inducible factor-1 were reduced by Hypoxia inducible factor-1 antisense oligonuclecotide. The findings suggested that antisense technique targeting Hypoxia inducible factor-1 might be an effective gene therapy of human hepatocellular carcinoma.


Antisense oligonucleotide Hypoxia inducible factor-1 Inhibitory effect Hepatocellular carcinoma Cell culture 



The health department of Zhejiang Province, China, funded the study.


  1. 1.
    Höckel M, Vaupel P. Tumor hypoxia: definitions and current clinical, biologic, and molecular aspects. J Natl Cancer Inst 2001;93(4):266–76.PubMedCrossRefGoogle Scholar
  2. 2.
    Schofield CJ, Ratcliffe PJ. Oxygen sensing by HIF hydroxylases. Nat Rev Mol Cell Biol 2004;5(5):343–54.PubMedCrossRefGoogle Scholar
  3. 3.
    Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer 2003;3(10):721–32.PubMedCrossRefGoogle Scholar
  4. 4.
    Belozerov VE, Van Meir EG. Hypoxia inducible factor-1: a novel target for cancer therapy. Anticancer Drugs 2005;16(9):901–909.PubMedCrossRefGoogle Scholar
  5. 5.
    Aboul-Fadl T. Antisense oligonucleotides: the state of the art. Curr Med Chem 2005;12(19):2193–214.PubMedCrossRefGoogle Scholar
  6. 6.
    Gleave ME, Monia BP. Antisense therapy for cancer. Nat Rev Cancer 2005;5(6):468–79.PubMedCrossRefGoogle Scholar
  7. 7.
    Peyman A, Helsberg M, et al. Inhibition of viral growth by antisense oligonucleotides directed against the IE110 and the UL30 mRNA of herpes simplex virus type-1. Biol Chem Hoppe Seyler 1995;376(3):195–8.PubMedGoogle Scholar
  8. 8.
    Monia BP, Johnston JF, Geiger T, Muller M, Fabbro D. Antitumor activity of a phosphorothioate antisense oligodeoxynucleotide targeted against C-raf kinase. Nat Med 1996;2(6):668–75.PubMedCrossRefGoogle Scholar
  9. 9.
    Brahimi-Horn MC, Pouysségur J. The hypoxia-inducible factor and tumor progression along the angiogenic pathway. Int Rev Cytol 2005;242:157–213.PubMedCrossRefGoogle Scholar
  10. 10.
    Pugh CW, Ratcliffe PJ. Regulation of angiogenesis by hypoxia: role of the HIF system. Nat Med 2003;9(6):677–84.PubMedCrossRefGoogle Scholar
  11. 11.
    Zagzag D, Zhong H, Scaizitti JM, et al. Expression of hypoxia-inducible factor 1alpha in brain tumors: association with angiogenesis, invasion, and progression. Cancer 2000;88(11):2606–18.PubMedCrossRefGoogle Scholar
  12. 12.
    Akakura N, Kobayashi M, Horiuchi I, et al. Constitutive expression of hypoxia-inducible factor-1alpha renders pancreatic cancer cells resistant to apoptosis induced by hypoxia and nutrient deprivation. Cancer Res 2001;61(17):6548–54.PubMedGoogle Scholar
  13. 13.
    Zhong H, Chiles K, Feldser D, et al. Modulation of hypoxia-inducible factor 1alpha expression by the epidermal growth factor/phosphatidylinositol 3-kinase/PTEN/AKT/FRAP pathway in human prostate cancer cells: implications for tumor angiogenesis and therapeutics. Cancer Res 2000;60(6):1541–5.PubMedGoogle Scholar
  14. 14.
    Zhang Q, Zhang ZF, Rao JY, et al. Treatment with siRNA and antisense oligonucleotides targeted to HIF-1alpha induced apoptosis in human tongue squamous cell carcinomas. Int J Cancer 2004;111(6):849–57.PubMedCrossRefGoogle Scholar
  15. 15.
    Zhang W, Zhang H, Xing L. Antisense oligonucleotide of hypoxia-inducible factor-1alpha suppresses growth and tumorigenicity of lung cancer cells A549. J Huazhong Univ Sci Technolog Med Sci 2006;26(4):448–50.PubMedCrossRefGoogle Scholar
  16. 16.
    Ding L, Chen X, Jing K, Wang H, Zhang W. Inhibition of the VEGF expression and cell growth in hepatocellular carcinoma by blocking HIF-1alpha and Smad3 binding site in VEGF promoter. J Huazhong Univ Sci Technolog Med Sci 2006;26(1):75–8.PubMedCrossRefGoogle Scholar
  17. 17.
    Vaupel P, Mayer A. Hypoxia and anemia: effects on tumor biology and treatment resistance. Transfus Clin Biol 2005;12(1):5–l0.PubMedCrossRefGoogle Scholar
  18. 18.
    Harrison L, Blackwell K. Hypoxia and anemia: factors in decreased sensitivity to radiation therapy and chemotherapy. Oncologist 2004;9(5):31–40.PubMedCrossRefGoogle Scholar
  19. 19.
    Ji Z, Yang G, Susan S, et al. Induction of hypoxia-inducible factor-1alpha overexpression by cobalt chloride enhances cellular resistance to photodynamic therapy. Cancer Letters 2006;244(2):182–9.PubMedCrossRefGoogle Scholar
  20. 20.
    Krishnamachary B, Berg-Dixon S, Kelly B, et al. Regulation of colon carcinoma cell invasion by hypoxia-inducible factor-1. Cancer Res 2003;63(5):1138–43.PubMedGoogle Scholar
  21. 21.
    Pennachietti S, Michieli P, Galluzzo M, et al. Hypoxia promotes invasive growth by transcriptional activation of the met protooncogene. Cancer Cell 2003;3(4):347–61.CrossRefGoogle Scholar
  22. 22.
    Birner P, Schindi M, Obermair A, et al. Overexpression of hypoxia-inducible factor 1alpha is a marker for an unfavorable prognosis in early-stage invasive cervical cancer. Cancer Res 2000;60(17):4693–6.PubMedGoogle Scholar
  23. 23.
    Birner P, Schindl M, Obermair A, Breitenecker G, Oberhuber G. Expression of hypoxia-inducible factor 1alpha in epithelial ovarian tumors: its impact on prognosis and on response to chemotherapy. Clin Cancer Res 2001;7(6):1661–8.PubMedGoogle Scholar
  24. 24.
    Bos R, Zhong H, Hanrahan CF, et al. Levels of hypoxia-inducible factor-1 alpha during breast carcinogenesis. J Natl Cancer Inst 2001;93(4):309–14.PubMedCrossRefGoogle Scholar
  25. 25.
    Aebersold DM, Burri P, Beer KT, et al. Expression of hypoxia-inducible factor-1alpha: a novel predictive and prognostic parameter in the radiotherapy of oropharyngeal cancer. Cancer Res 2001;61(7):2911–6.PubMedGoogle Scholar
  26. 26.
    Koukourakis MI, Giatromanolaki A, Skarlatos J, et al. Hypoxia inducible factor (HIF-1α and HIF-2α) expression in early esophageal cancer and response to photodynamic therapy and radiotherapy. Cancer Res 2001;61(5):1830–2.PubMedGoogle Scholar
  27. 27.
    Powis G, Kirkpatrick L. Hypoxia inducible factor-1α as a cancer drug target. Mol Cancer Ther 2004;3(5):647–54.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Chen WeiXing
    • 1
  • Hu Tiantian
    • 1
  • Ni Qun
    • 1
  • Yu Chaohui
    • 1
  • Xu Ping
    • 1
  1. 1.Department of Gastroenterology, First Affiliated HospitalMedical College of Zhejiang UniversityHangzhouChina

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