Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Basic Research

Cyclin-dependent kinase inhibitor, P276-00, inhibits HIF-1α and induces G2/M arrest under hypoxia in prostate cancer cells

Prostate Cancer and Prostatic Diseases volume 15, pages 15–27 (2012)Cite this article

Abstract

Background:

Hypoxia-inducible factor-1 (HIF-1) is a master regulator of the transcriptional response to oxygen deprivation and controls genes involved in glycolysis, angiogenesis, migration and invasion. Overexpression of HIF-1α has been demonstrated in many common human cancers.

Methods:

Luciferase reporter gene assay under hypoxia and normoxia was used to demonstrate transcriptional inhibition of HIF-1 by P276-00. Detailed studies such as western blotting, reverse-transcriptase-PCR and immunofluorescence were carried out to elucidate its mechanism of action. Cytotoxic potential of P276-00 under normoxia and hypoxia was determined on prostate cancer cells using CCK-8 assay, and cell-cycle analysis was carried out using flow cytometry. Antiangiogenic activity of P276-00 was demonstrated by migration assay and tube-formation assay. Efficacy study of P276-00 was performed in a PC-3 xenograft model.

Results:

P276-00 inhibits transcriptional activation of HIF-1 under hypoxia. It suppressed hypoxia-mediated nuclear HIF-1α expression, as well as phosphorylation of Akt and 4E-BP1 and abrogated expression of HIF-1-inducible gene viz. vascular endothelial growth factor. Under hypoxia, P276-00 did not exhibit enhanced cytotoxic activity in prostate cancer cells but arrested them in the G2/M phase of the cell cycle. The tubular formation of human umbilical vein endothelial cells and migration of prostate cancer cells were also inhibited by P276-00 in vitro. In addition, it demonstrated significant in vivo efficacy in the PC-3 xenograft model.

Conclusions:

Given its low toxicity profile, its demonstrated antitumor activity and its potential to inhibit the HIF-1 pathway, P276-00 should be considered as antiangiogenic chemotherapy for prostate cancer.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  1. Sun Y-L, Liu Y-N, Huang Y-T, Pan S-L, Huang D-Y, Guh J-H et al. YC-1 inhibits HIF-1 expression in prostate cancer cells: contribution of Akt/NF-kB signaling to HIF-1α accumulation during hypoxia. Oncogene 2007; 26: 3941–3951.

    Article  CAS  PubMed  Google Scholar 

  2. Akakura N, Kobayashi M, Horiuchi I, Suzuki A, Wang J, Chen J et al. Constitutive expression of hypoxia-inducible factor-1α renders pancreatic cancer cells resistant to apoptosis induced by hypoxia and nutrient deprivation. Cancer Res 2001; 61: 6548–6554.

    CAS  PubMed  Google Scholar 

  3. Zhong H, De Marzo AM, Laughner E, Lim M, Hilton DA, Zagzag D et al. Overexpression of hypoxia-inducible factor 1α in common human cancers and their metastases. Cancer Res 1999; 59: 5830–5835.

    CAS  PubMed  Google Scholar 

  4. Newcomb EW, Lukyanov Y, Schnee T, Ali MA, Lan L, Zagzag D . Noscapine inhibits hypoxia-mediated HIF-1alpha expression and angiogenesis in vitro: a novel function for an old drug. Int J Oncol 2006; 28: 1121–1130.

    CAS  PubMed  Google Scholar 

  5. Sommani P, Yamashita K, Miyoshi T, Tsunemine H, Kodaki T, Mori H et al. Inhibitory effect of 6-Formylpterin on HIF-1α protein accumulation. Biol Pharm Bull 2007; 30: 2181–2184.

    Article  CAS  PubMed  Google Scholar 

  6. Dong-Li Z, Liu L-Z, Shi X, Fang J, Jiang B-H . Benzo[α] pyrene-3,6-dione inhibited VEGF expression through inducing HIF-1α degradation. Biochem Biophys Res Commun 2007; 357: 517–523.

    Article  Google Scholar 

  7. Bilton RL, Wilson WR . Exploiting tumour hypoxia in cancer treatment. Nat Rev Cancer 2003; 4: 437–447.

    Google Scholar 

  8. Zhong H, Chiles K, Feldser D, Laughner E, Hanrahan C, Georgescu MM 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: 1541–1545.

    CAS  PubMed  Google Scholar 

  9. Laughner E, Taghavi P, Chiles K, Mahon PC, Semenza GL . HER2 (neu) signaling increases the rate of hypoxia-inducible factor 1alpha (HIF1-alpha) synthesis: novel mechanism for HIF-1-mediated vascular endothelial growth factor expression. Mol Cell Biol 2001; 21: 3995–4004.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Treins C, Giorgetti-Peraldi S, Murdaca J, Semenza GL, Van OE . Insulin stimulates hypoxia-inducible factor 1 through a phosphotidyl 3-kinase/target of rapamycin-dependent signaling pathway. J Biol Chem 2002; 277: 27975–27981.

    Article  CAS  PubMed  Google Scholar 

  11. Joshi KS, Rathos MJ, Joshi RD, Sivakumar M, Mascarenhas M, Kamble S et al. In vitro antitumor properties of a novel cyclin-dependent kinase inhibitor, P276-00-00. Mol Cancer Ther 2007; 6: 918–925.

    Article  CAS  PubMed  Google Scholar 

  12. Joshi KS, Rathos MJ, Mahajan P, Wagh V, Shenoy S, Chile S et al. P276-00-00, a novel cyclin-dependent inhibitor induces G1-G2 arrest, shows antitumor activity on cisplatin-resistant cells and significant in vivo efficacy in tumor models. Mol Cancer Ther 2007; 6: 926–934.

    Article  CAS  PubMed  Google Scholar 

  13. Rapisarda A, Uranchimeg B, Scudiero DA, Selby M, Sausville EA, Shoemaker RH et al. Identification of small molecule inhibitors of hypoxia-inducible factor 1 transcriptional activation pathways. Cancer Res 2002; 62: 4316–4324.

    CAS  PubMed  Google Scholar 

  14. Zundel W, Schindler C, Haas-Kogan D, Koong A, Kaper F, Chen E . Loss of PTEN facilitates HIF-1-mediated gene expression. Genes Dev 2000; 14: 391–396.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Chen EY, Mazure NM, Cooper JA, Giaccia AJ . Hypoxia activates a platelet-derived growth factor receptor/phosphatidylinositol 3-kinase/Akt pathway that results in glycogen synthase kinase-3 inactivation. Cancer Res 2001; 61: 2429–2433.

    CAS  PubMed  Google Scholar 

  16. Hudson CC, Liu M, Chiang GG, Otterness DM, Loomis DC, Kaper F et al. Regulation of hypoxia-inducible factor 1alpha expression and function by the mammalian target of rapamycin. Mol Cell Biol 2002; 22: 7004–7014.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Li M-H, Miao Z-H, Tan W-F, Yue J-M, Zhang X-W, Ding J . Pseudolaric acid B inhibits angiogenesis and reduces hypoxia-inducible factor 1α by promoting proteasome-mediated degradation. Clin Cancer Res 2004; 10: 8266–8274.

    Article  CAS  PubMed  Google Scholar 

  18. Kong D, Park E-J, Stephen AG, Calvani M, Cardellina JH, Monks A et al. Echinomycin, a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity. Cancer Res 2005; 65: 9047–9055.

    Article  CAS  PubMed  Google Scholar 

  19. Sun X, Kanwar JR, Leung E, Lehnert K, Wang D, Krissansen GW . Gene transfer of antisense hypoxia inducible factor-1α enhances the therapeutic efficacy of cancer immunotherapy. Gene Ther 2001; 8: 638–645.

    Article  CAS  PubMed  Google Scholar 

  20. Zhang Q, Zhang ZF, Rao JY, Sato JD, Brown J, Messadi DV et al. Treatment with siRNA and antisense oligonucleotides targeted to HIF- 1α induced apoptosis in human tongue squamous cell carcinomas. Int J Cancer 2004; 111: 849–857.

    Article  CAS  PubMed  Google Scholar 

  21. Neckers L . Hsp90 inhibitors as novel cancer chemotherapeutic agents. Trends Mol Med 2002; 8: S55–S61.

    Article  CAS  PubMed  Google Scholar 

  22. Mabjeesh NJ, Post DE, Willard MT, Kaur B, Van Meir EG, Simons JW et al. Geldanamycin induces degradation of hypoxia-inducible factor 1α protein via the proteosome pathway in prostate cancer cells. Cancer Res 2002; 62: 2478–2482.

    CAS  PubMed  Google Scholar 

  23. Kurebayashi J, Otsuki T, Kurosumi M, Soga S, Akinga S, Sonoo H . A radicicol derivative, KF58333, inhibits expression of hypoxia-inducible factor-1α and vascular endothelial growth factor, angiogenesis and growth of human breast cancer xenografts. J Cancer Res 2001; 92: 1342–1351.

    CAS  Google Scholar 

  24. Welsh SJ, Williams RR, Birmingham A, Newman DJ, Kirkpatrick DL, Powis G . The thioredoxin redox inhibitors 1-methylpropyl 2-imidazolyl disulfide and pleurotin inhibit hypoxia-induced factor 1a and vascular endothelial growth factor formation. Mol Cancer Ther 2003; 2: 235–243.

    CAS  PubMed  Google Scholar 

  25. Tan C, de Noronha RG, Roecker AJ, Pyrzynska B, Khwaja F, Zhang Z et al. Identification of a novel small-molecule inhibitor of the hypoxia inducible factor 1 pathway. Cancer Res 2005; 65: 605–612.

    CAS  PubMed  Google Scholar 

  26. Mabjeesh NJ, Willard MT, Harris WB, Sun HY, Wang R, Thong H et al. Dibenzoylmethane, a natural dietary compound, induces HIF- 1α and increases expression of VEGF. Biochem Biophys Res Commun 2003; 303: 279–286.

    Article  CAS  PubMed  Google Scholar 

  27. Rapisarda A, Uranchimeg B, Sordet O, Pommier Y, Shoemaker RH, Melillo G . Topoisomerase I-mediated inhibition of hypoxia-inducible factor 1: mechanism and therapeutic implications. Cancer Res 2004; 64: 1475–1482.

    Article  CAS  PubMed  Google Scholar 

  28. Yeo EJ, Chun YS, Cho YS, Kim J, Lee J-C, Kim M-S et al. YC-1: a potential anticancer drug targeting hypoxia-inducible factor 1. J Natl Cancer Inst 2003; 95: 516–525.

    Article  CAS  PubMed  Google Scholar 

  29. Welsh S, Williams R, Kirkpatrick L, Paine-Murrieta G, Powis G . Antitumor activity and pharmacodynamic properties of PX-478, an inhibitor of hypoxia-inducible factor-1α. Mol Cancer Ther 2004; 3: 233–244.

    Article  CAS  PubMed  Google Scholar 

  30. Giaccia A, Siim BG, Johnson RS . HIF-1 as a target for drug development. Nat Rev Drug Discov 2003; 2: 803–811.

    Article  CAS  PubMed  Google Scholar 

  31. Semenza GL . Targeting HIF-1 for cancer therapy. Nat Rev Cancer 2003; 3: 721–732.

    Article  CAS  PubMed  Google Scholar 

  32. Powis G, Kirkpatrick L . Hypoxia inducible factor- 1alpha as a cancer drug target. Mol Cancer Ther 2004; 3: 647–654.

    CAS  PubMed  Google Scholar 

  33. Yeo EJ, Chun YS, Park JW . New anticancer strategies targeting HIF-1. Biochem Pharmacol 2004; 68: 1061–1069.

    Article  CAS  PubMed  Google Scholar 

  34. Belozerov VE, Van Meir EG . Hypoxia inducible factor-1: a novel target for cancer therapy. Anticancer Drugs 2005; 16: 901–909.

    Article  CAS  PubMed  Google Scholar 

  35. Escuin D, Kline ER, Giannakakou P . Both microtubule-stabilizing and microtubule-destabilizing drugs inhibit hypoxia-inducible factor-1alpha accumulation and activity by disrupting microtubule function. Cancer Res 2005; 65: 9021–9028.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Wiedmann MW, Caca K . Molecularly targeted therapy for gastrointestinal cancer. Curr Cancer Drug Targets 2005; 5: 171–193.

    Article  CAS  PubMed  Google Scholar 

  37. Generali D, Berruti A, Brizzi MP, Campo L, Bonardi S, Wigfield S et al. Hypoxia-inducible factor-1α expression predicts a poor response to primary chemoendocrine therapy and disease-free survival in primary human breast cancer. Clin Cancer Res 2006; 12: 4562–4568.

    Article  CAS  PubMed  Google Scholar 

  38. Patiar S, Harris AL . Role of hypoxia-inducible factor-1α as a cancer therapy target. Endocr Relat Cancer 2006; 13: 61–75.

    Article  Google Scholar 

  39. Powis G, Kirkpatrick L . Hypoxia inducible factor-1α as a cancer drug target. Mol Cancer Ther 2004; 5: 647–654.

    Google Scholar 

  40. Zhong H, Semenza GL, Simons JW, De Marzo AM . Up-regulation of hypoxia-inducible factor 1alpha is an early event in prostate carcinogenesis. Cancer Detect Prev 2004; 28: 88–93.

    Article  CAS  PubMed  Google Scholar 

  41. Du Z, Fujiyama C, Chen Y, Masaki Z . Expression of hypoxia-inducible factor 1alpha in human normal, benign, and malignant prostate tissue. Chin Med J 2003; 116: 1936–1939.

    CAS  PubMed  Google Scholar 

  42. Fang J, Ding M, Yang L, Liu L-Z, Jiang B-H . PI3K/PTEN/AKT signaling regulates prostate tumor angiogenesis. Cell Signal 2007; 19: 2487–2497.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Karni R, Dor Y, Keshet E, Meyuhas O, Levitzki A . Activated pp60c-Src leads to elevated hypoxia-inducible factor (HIF)-1α expression under normoxia. J Biol Chem 2002; 277: 42919–42925.

    Article  CAS  PubMed  Google Scholar 

  44. Lou J, Zhou X, Weng Q, Wang D-d, Xia Q, Hu Y et al. XQ2, a novel TPZ derivative, induced G2/M arrest and apoptosis under hypoxia in non-small cell lung cancer cells. Biosci Biotechnol Biochem 2010; 74: 1181–1187.

    Article  CAS  PubMed  Google Scholar 

  45. Pauli RM, Strauss BS . Proliferation of human peripheral lymphocytes. Characteristics of cells once stimulated or restimulated by concanavalin A. Exp Cell Res 1973; 82: 357–366.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The study has been supported and carried out at Piramal Life Sciences, Goregaon, Mumbai, India. We extend our thanks for the support.

Author information

Authors and Affiliations

  1. Oncology Franchisee, Cancer Pharmacology, Piramal Life Sciences, Mumbai, Maharashtra, India

    S M Manohar, A A Padgaonkar, A Jalota-Badhwar & S V Rao

  2. Target Identification Group, Piramal Life Sciences, Mumbai, Maharashtra, India

    K S Joshi

Authors
  1. S M Manohar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A A Padgaonkar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Jalota-Badhwar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S V Rao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K S Joshi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K S Joshi.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Manohar, S., Padgaonkar, A., Jalota-Badhwar, A. et al. Cyclin-dependent kinase inhibitor, P276-00, inhibits HIF-1α and induces G2/M arrest under hypoxia in prostate cancer cells. Prostate Cancer Prostatic Dis 15, 15–27 (2012). https://doi.org/10.1038/pcan.2011.51

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/pcan.2011.51

Keywords

This article is cited by

Search

Advanced search

Quick links