Clinical & Experimental Metastasis

, Volume 30, Issue 4, pp 381–392 | Cite as

Combined effect of dehydroxymethylepoxyquinomicin and gemcitabine in a mouse model of liver metastasis of pancreatic cancer

  • Keiichi Suzuki
  • Koichi Aiura
  • Sachiko Matsuda
  • Osamu Itano
  • Osamu Takeuchi
  • Kazuo Umezawa
  • Yuko Kitagawa
Research Paper

Abstract

Activation of nuclear factor-κB (NF-κB) has been implicated in metastasis of pancreatic cancer. We investigated the effects of the novel NF-κB inhibitor dehydroxymethylepoxyquinomicin (DHMEQ) on the inhibition of liver metastasis of pancreatic cancer in a mouse model of clinical liver metastasis. Nude mice were xenografted by intra-portal-vein injection with the human pancreatic adenocarcinomas cell line AsPC-1 via small laparotomy. Mice were treated with DHMEQ and gemcitabine (GEM), alone or in combination. The combination of GEM + DHMEQ showed a stronger antitumor effect than either monotherapy. Apoptosis induction in the metastatic foci was greatest in the DHMEQ + GEM group. Significant reductions in the numbers of neovessels were also seen in the DHMEQ and/or GEM groups. Cell growth inhibition assays revealed no synergistic effect of combination therapy, although each monotherapy had an individual cytotoxic effect. Combination therapy produced the greatest inhibition of tumor cell invasiveness in chemoinvasion assay. In addition, combination therapy significantly down-regulated the expression level of matrix metalloproteinase (MMP)-9 mRNA in AsPC-1 cells. DHMEQ also markedly down-regulated interleukin-8 and MMP-9, while GEM caused moderate down-regulation of vascular endothelial growth factor in metastatic foci, demonstrated by quantitative reverse transcription-polymerase chain reaction. These results demonstrate that DHMEQ can exert anti-tumor effects by inhibiting angiogenesis and tumor cell invasion, and by inducing apoptosis. Combination therapy with DHMEQ and GEM also showed potential efficacy. DHMEQ is a promising drug for the treatment of advanced pancreatic cancer.

Keywords

Pancreatic cancer NF-kappaB DHMEQ Liver metastasis Anticancer effect 

Abbrevations

NF-κB

Nuclear factor-κB

DHMEQ

Dehydroxymethylepoxyquinomicin

CMC

Chrolomethylcellulose

FBS

Fetal bovine serum

PBS

Phosphate-buffered saline

MVD

Microvessel density

VEGF

Vascular endothelial growth factor

MMP

Matrix metalloproteinase

IL-8

Interleukin-8

References

  1. 1.
    Heinemann V (2002) Present and future treatment of pancreatic cancer. Semin Oncol 29(3 Suppl 9):23–31PubMedCrossRefGoogle Scholar
  2. 2.
    Haller DG (2002) Future directions in the treatment of pancreatic cancer. Semin Oncol 29(6 Suppl 20):31–39PubMedGoogle Scholar
  3. 3.
    Jemal A et al (2009) Cancer statistics, 2009. CA Cancer J Clin 59(4):225–249PubMedCrossRefGoogle Scholar
  4. 4.
    Cullinan SA et al (1985) A comparison of three chemotherapeutic regimens in the treatment of advanced pancreatic and gastric carcinoma. Fluorouracil vs fluorouracil and doxorubicin vs fluorouracil, doxorubicin, and mitomycin. JAMA 253(14):2061–2067PubMedCrossRefGoogle Scholar
  5. 5.
    DeCaprio JA et al (1991) Fluorouracil and high-dose leucovorin in previously untreated patients with advanced adenocarcinoma of the pancreas: results of a phase II trial. J Clin Oncol 9(12):2128–2133PubMedGoogle Scholar
  6. 6.
    Lionetto R et al (1995) No standard treatment is available for advanced pancreatic cancer. Eur J Cancer 31A(6):882–887PubMedCrossRefGoogle Scholar
  7. 7.
    Burris HA 3rd et al (1997) Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer: a randomized trial. J Clin Oncol 15(6):2403–2413PubMedGoogle Scholar
  8. 8.
    Oettle H et al (2007) Adjuvant chemotherapy with gemcitabine vs observation in patients undergoing curative-intent resection of pancreatic cancer: a randomized controlled trial. JAMA 297(3):267–277PubMedCrossRefGoogle Scholar
  9. 9.
    Sen R, Baltimore D (1986) Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell 46(5):705–716PubMedCrossRefGoogle Scholar
  10. 10.
    Cogswell PC, Scheinman RI, Baldwin AS Jr (1993) Promoter of the human NF-kappa B p50/p105 gene. Regulation by NF-kappa B subunits and by c-REL. J Immunol 150(7):2794–2804PubMedGoogle Scholar
  11. 11.
    Baldwin AS Jr (1996) The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol 14:649–683PubMedCrossRefGoogle Scholar
  12. 12.
    Wang CY, Mayo MW, Baldwin AS Jr (1996) TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-kappaB. Science 274(5288):784–787PubMedCrossRefGoogle Scholar
  13. 13.
    Pikarsky E et al (2004) NF-kappaB functions as a tumour promoter in inflammation-associated cancer. Nature 431(7007):461–466PubMedCrossRefGoogle Scholar
  14. 14.
    Redell MS, Tweardy DJ (2005) Targeting transcription factors for cancer therapy. Curr Pharm Des 11(22):2873–2887PubMedCrossRefGoogle Scholar
  15. 15.
    Dolcet X et al (2005) NF-kB in development and progression of human cancer. Virchows Arch 446(5):475–482PubMedCrossRefGoogle Scholar
  16. 16.
    Karin M, Greten FR (2005) NF-kappaB: linking inflammation and immunity to cancer development and progression. Nat Rev Immunol 5(10):749–759PubMedCrossRefGoogle Scholar
  17. 17.
    Ariga A et al (2002) Inhibition of tumor necrosis factor-alpha-induced nuclear translocation and activation of NF-kappa B by dehydroxymethylepoxyquinomicin. J Biol Chem 277(27):24625–24630PubMedCrossRefGoogle Scholar
  18. 18.
    Watanabe M et al (2005) A novel NF-kappaB inhibitor DHMEQ selectively targets constitutive NF-kappaB activity and induces apoptosis of multiple myeloma cells in vitro and in vivo. Int J Cancer 114(1):32–38PubMedCrossRefGoogle Scholar
  19. 19.
    Poma P et al (2006) Antitumor effects of the novel NF-kappaB inhibitor dehydroxymethyl-epoxyquinomicin on human hepatic cancer cells: analysis of synergy with cisplatin and of possible correlation with inhibition of pro-survival genes and IL-6 production. Int J Oncol 28(4):923–930PubMedGoogle Scholar
  20. 20.
    Ohsugi T et al (2006) In vitro and in vivo antitumor activity of the NF-kappaB inhibitor DHMEQ in the human T-cell leukemia virus type I-infected cell line, HUT-102. Leuk Res 30(1):90–97PubMedCrossRefGoogle Scholar
  21. 21.
    Matsumoto N et al (2000) Synthesis of NF-kappaB activation inhibitors derived from epoxyquinomicin C. Bioorg Med Chem Lett 10(9):865–869PubMedCrossRefGoogle Scholar
  22. 22.
    Wang W et al (1999) The nuclear factor-kappa B RelA transcription factor is constitutively activated in human pancreatic adenocarcinoma cells. Clin Cancer Res 5(1):119–127PubMedGoogle Scholar
  23. 23.
    Sclabas GM et al (2003) Restoring apoptosis in pancreatic cancer cells by targeting the nuclear factor-kappaB signaling pathway with the anti-epidermal growth factor antibody IMC-C225. J Gastrointest Surg 7(1): 37–43; discussionGoogle Scholar
  24. 24.
    Liptay S et al (2003) Mitogenic and antiapoptotic role of constitutive NF-kappaB/Rel activity in pancreatic cancer. Int J Cancer 105(6):735–746PubMedCrossRefGoogle Scholar
  25. 25.
    Liu LP et al (2010) The role of NF-kappaB in Hepatitis b virus X protein-mediated upregulation of VEGF and MMPs. Cancer Invest 28(5):443–451PubMedCrossRefGoogle Scholar
  26. 26.
    Lauricella-Lefebvre MA et al (1993) Stimulation of the 92-kD type IV collagenase promoter and enzyme expression in human melanoma cells. Invasion Metastasis 13(6):289–300PubMedGoogle Scholar
  27. 27.
    Griffin JF et al (1990) Patterns of failure after curative resection of pancreatic carcinoma. Cancer 66(1):56–61PubMedCrossRefGoogle Scholar
  28. 28.
    Weidner N (1995) Current pathologic methods for measuring intratumoral microvessel density within breast carcinoma and other solid tumors. Breast Cancer Res Treat 36(2):169–180PubMedCrossRefGoogle Scholar
  29. 29.
    Chen WH et al (1982) Human pancreatic adenocarcinoma: in vitro and in vivo morphology of a new tumor line established from ascites. In Vitro 18(1):24–34PubMedCrossRefGoogle Scholar
  30. 30.
    Fujioka S et al (2003) Function of nuclear factor kappaB in pancreatic cancer metastasis. Clin Cancer Res 9(1):346–354PubMedGoogle Scholar
  31. 31.
    Loukopoulos P et al (2004) Orthotopic transplantation models of pancreatic adenocarcinoma derived from cell lines and primary tumors and displaying varying metastatic activity. Pancreas 29(3):193–203PubMedCrossRefGoogle Scholar
  32. 32.
    Tan MH, Chu TM (1985) Characterization of the tumorigenic and metastatic properties of a human pancreatic tumor cell line (AsPC-1) implanted orthotopically into nude mice. Tumour Biol 6(1):89–98PubMedGoogle Scholar
  33. 33.
    Watanabe M et al (2005) Dual targeting of transformed and untransformed HTLV-1-infected T cells by DHMEQ, a potent and selective inhibitor of NF-kappaB, as a strategy for chemoprevention and therapy of adult T-cell leukemia. Blood 106(7):2462–2471PubMedCrossRefGoogle Scholar
  34. 34.
    Yamamoto M et al (2008) Inactivation of NF-kappaB components by covalent binding of (−)-dehydroxymethylepoxyquinomicin to specific cysteine residues. J Med Chem 51(18):5780–5788PubMedCrossRefGoogle Scholar
  35. 35.
    Shimada C et al (2010) Efficient cellular uptake of the novel NF-kappaB inhibitor (−)-DHMEQ and irreversible inhibition of NF-kappaB in neoplastic cells. Oncol Res 18(11–12):529–535PubMedCrossRefGoogle Scholar
  36. 36.
    Gilmore TD (1999) The Rel/NF-kappaB signal transduction pathway: introduction. Oncogene 18(49):6842–6844PubMedCrossRefGoogle Scholar
  37. 37.
    Pahl HL (1999) Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene 18(49):6853–6866PubMedCrossRefGoogle Scholar
  38. 38.
    Karin M (1999) The beginning of the end: IkappaB kinase (IKK) and NF-kappaB activation. J Biol Chem 274(39):27339–27342PubMedCrossRefGoogle Scholar
  39. 39.
    Gilroy DW et al (2004) Inflammatory resolution: new opportunities for drug discovery. Nat Rev Drug Discov 3(5):401–416PubMedCrossRefGoogle Scholar
  40. 40.
    Maeda S et al (2005) Nod2 mutation in Crohn’s disease potentiates NF-kappaB activity and IL-1beta processing. Science 307(5710):734–738PubMedCrossRefGoogle Scholar
  41. 41.
    Arkan MC et al (2005) IKK-beta links inflammation to obesity-induced insulin resistance. Nat Med 11(2):191–198PubMedCrossRefGoogle Scholar
  42. 42.
    Chandler NM, Canete JJ, Callery MP (2004) Increased expression of NF-kappa B subunits in human pancreatic cancer cells. J Surg Res 118(1):9–14PubMedCrossRefGoogle Scholar
  43. 43.
    Sclabas GM et al (2005) Nuclear factor kappa B activation is a potential target for preventing pancreatic carcinoma by aspirin. Cancer 103(12):2485–2490PubMedCrossRefGoogle Scholar
  44. 44.
    Matsumoto N et al (1997) Epoxyquinomicins A, B, C and D, new antibiotics from Amycolatopsis. II. Effect on type II collagen-induced arthritis in mice. J Antibiot (Tokyo) 50(11):906–911CrossRefGoogle Scholar
  45. 45.
    Ohsugi T et al (2005) In vivo antitumor activity of the NF-kappaB inhibitor dehydroxymethylepoxyquinomicin in a mouse model of adult T-cell leukemia. Carcinogenesis 26(8):1382–1388PubMedCrossRefGoogle Scholar
  46. 46.
    Ohsugi T et al (2007) Dehydroxymethylepoxyquinomicin (DHMEQ) therapy reduces tumor formation in mice inoculated with tax-deficient adult T-cell leukemia-derived cell lines. Cancer Lett 257(2):206–215PubMedCrossRefGoogle Scholar
  47. 47.
    Watanabe M et al (2007) IkappaBalpha independent induction of NF-kappaB and its inhibition by DHMEQ in Hodgkin/Reed-Sternberg cells. Lab Invest 87(4):372–382PubMedGoogle Scholar
  48. 48.
    Starenki DV et al (2004) Induction of thyroid cancer cell apoptosis by a novel nuclear factor kappaB inhibitor, dehydroxymethylepoxyquinomicin. Clin Cancer Res 10(20):6821–6829PubMedCrossRefGoogle Scholar
  49. 49.
    Matsumoto G et al (2005) Targeting of nuclear factor kappaB pathways by dehydroxymethylepoxyquinomicin, a novel inhibitor of breast carcinomas: antitumor and antiangiogenic potential in vivo. Clin Cancer Res 11(3):1287–1293PubMedGoogle Scholar
  50. 50.
    Tatetsu H et al (2005) Dehydroxymethylepoxyquinomicin, a novel nuclear factor-kappaB inhibitor, induces apoptosis in multiple myeloma cells in an IkappaBalpha-independent manner. Mol Cancer Ther 4(7):1114–1120PubMedCrossRefGoogle Scholar
  51. 51.
    Umezawa K (2006) Inhibition of tumor growth by NF-kappaB inhibitors. Cancer Sci 97(10):990–995PubMedCrossRefGoogle Scholar
  52. 52.
    Matsumoto G et al (2005) Enhancement of the caspase-independent apoptotic sensitivity of pancreatic cancer cells by DHMEQ, an NF-kappaB inhibitor. Int J Oncol 27(5):1247–1255PubMedGoogle Scholar
  53. 53.
    Hu DE, Hori Y, Fan TP (1993) Interleukin-8 stimulates angiogenesis in rats. Inflammation 17(2):135–143PubMedCrossRefGoogle Scholar
  54. 54.
    Strieter RM et al (1992) Interleukin-8. A corneal factor that induces neovascularization. Am J Pathol 141(6):1279–1284PubMedGoogle Scholar
  55. 55.
    Olive KP et al (2009) Inhibition of Hedgehog signaling enhances delivery of chemotherapy in a mouse model of pancreatic cancer. Science 324(5933):1457–1461PubMedCrossRefGoogle Scholar
  56. 56.
    Bruns CJ et al (2002) Effect of the vascular endothelial growth factor receptor-2 antibody DC101 plus gemcitabine on growth, metastasis and angiogenesis of human pancreatic cancer growing orthotopically in nude mice. Int J Cancer 102(2):101–108PubMedCrossRefGoogle Scholar
  57. 57.
    Jia L et al (2005) Antiangiogenic therapy for human pancreatic carcinoma xenografts in nude mice. World J Gastroenterol 11(3):447–450PubMedGoogle Scholar
  58. 58.
    Szlosarek PW, Balkwill FR (2003) Tumour necrosis factor alpha: a potential target for the therapy of solid tumours. Lancet Oncol 4(9):565–573PubMedCrossRefGoogle Scholar
  59. 59.
    Kaltschmidt B et al (2002) Cyclooxygenase-2 is a neuronal target gene of NF-kappaB. BMC Mol Biol 3:16PubMedCrossRefGoogle Scholar
  60. 60.
    Nakanishi C, Toi M (2005) Nuclear factor-kappaB inhibitors as sensitizers to anticancer drugs. Nat Rev Cancer 5(4):297–309PubMedCrossRefGoogle Scholar
  61. 61.
    Jazirehi AR et al (2005) Rituximab (chimeric anti-CD20 monoclonal antibody) inhibits the constitutive nuclear factor-{kappa}B signaling pathway in non-Hodgkin’s lymphoma B-cell lines: role in sensitization to chemotherapeutic drug-induced apoptosis. Cancer Res 65(1):264–276PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Keiichi Suzuki
    • 1
  • Koichi Aiura
    • 2
  • Sachiko Matsuda
    • 3
  • Osamu Itano
    • 3
  • Osamu Takeuchi
    • 4
  • Kazuo Umezawa
    • 5
  • Yuko Kitagawa
    • 3
  1. 1.Department of SurgeryKitasato Institute HospitalTokyoJapan
  2. 2.Department of SurgeryKawasaki City HospitalKawasakiJapan
  3. 3.Department of SurgeryKeio University School of MedicineTokyoJapan
  4. 4.Biomedical LaboratoryKitasato Institute HospitalTokyoJapan
  5. 5.Department of Applied ChemistryFaculty of Science and Technology, Keio University, KanagawaYokohamaJapan

Personalised recommendations