Clinical & Experimental Metastasis

, Volume 21, Issue 5, pp 461–467 | Cite as

Cellular mechanisms mediating the anti-invasive properties of the ornithine decarboxylase inhibitor α-difluoromethylornithine (DFMO) in human breast cancer cells

  • Andrea Manni
  • Sharlene Washington
  • David Mauger
  • Deborah A. Hackett
  • Michael F. Verderame
Article

Abstract

We have shown that inhibition of polyamine biosynthesis with α-difluoromethylornithine (DFMO) reduces in vitro invasiveness and metastatic capacity of MDA-MB-435 breast cancer cells. These experiments investigated the mechanisms mediating the anti-invasive properties of DFMO. DFMO did not affect phosphorylation of FAK or Akt, but increased ERK phosphorylation by approximately threefold. To test the biologic significance of this finding, we tested the effect of the MEK inhibitor PD98059 on in vitro invasiveness of MDA-MB-435 breast cancer cells, both in the absence and in the presence of the proinvasive peptide hepatocyte growth factor (HGF) as a chemoattractant. We observed that PD98059 treatment reversed the anti-invasive effect of DFMO under both experimental conditions. Next, we tested the influence of DFMO on the production of the prometastatic peptide osteopontin (OPN) and the anti-metastatic protein thrombospondin-1 (TSP-1). DFMO treatment, while not affecting OPN production, markedly increased the TSP-1 level in the conditioned media. This effect was abolished by putrescine administration, thus indicating the specificity of the DFMO action through the polyamine pathway. PD98059 completely blocked the stimulatory effect of DFMO on TSP-1 production, which supports a mediatory role for activation of the MAPK pathway in the upregulation of this anti-metastatic peptide by DFMO. In summary, our results show that the increase in ERK phosphorylation induced by DFMO plays a critical role in the anti-invasive action of the drug and in its ability to upregulate TSP-1 production.

Keywords

breast cancer invasiveness polyamines signal transduction 

Abbreviation

ODC

ornithine decarboxylase

DFMO

α-difluromethylornithine

ERK

extracellular signal-regulated kinase

MAPK

mitogen-activated protein kinase

MEK-1

MAP/ERK kinase-1

OPN

osteopontin

TSP-1

thrombospondin 1

HGF

hepatocyte growth factor

FAK

focal adhesion kinase

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References

  1. Glikman, P, Manni, A, Demers, L, Bartholomew, M 1989Polyamine involvement in the growth of hormone-responsive and -resistant human breast cancer cells in culture.Clinical & Experimental Metastasis4913711376Google Scholar
  2. Kendra, KL, Katzenellenbogen, BS 1987An evaluation of the involvement of polyamines in modulating MCF-7 human breast cancer cell proliferation and progesterone receptor levels by estrogen and antiestrogen.Clinical & Experimental Metastasis28123128Google Scholar
  3. Manni, A, Grove, R, Kunselman, S, Aldaz, M 1995Involvement of the polyamine pathway in breast cancer progression.Clinical & Experimental Metastasis924957Google Scholar
  4. Manni, A, Mauger, D, Gimotty, P, Badger, B 1996Prognostic influence on survival of increased ornithine decarboxylase activity in human breast cancer.Clinical & Experimental Metastasis219011906Google Scholar
  5. Canizares, F, Salinas, J, las Heras, M,  et al. 1999Prognostic value of ornithine decarboxylase and polyamines in human breast cancer: correlation with clinicopathologic parameters.Clinical & Experimental Metastasis520352041Google Scholar
  6. Manni, A, Washington, S, Griffith, JW, Verderame, MF, Mauger, D, Demers, LM,  et al. 2002Influence of polyamines on in vitro and in vivo features of aggressive and metast behavior by human breast cancer cells.Clinical & Experimental Metastasis1995105Google Scholar
  7. Manni, A, Washington, S, Craig, L,  et al. 2003Effects of alpha-difluoromethylornithine on local recurrence and pulmonary metastasis from MDA-MB-435 breast cancer xenografts in nude mice.Clinical & Experimental Metastasis20321325Google Scholar
  8. Tuck, AB, Chambers, AF 2001The role of osteopontin in breast cancer: clinical and experimental studies.Clinical & Experimental Metastasis6419429Google Scholar
  9. Tuck, AB, Arsenault, DM, O’Malley, FP,  et al. 1999Osteopontin induces increased invasiveness and plasminogen activator expression of human mammary epithelial cells.Clinical & Experimental Metastasis1842374246Google Scholar
  10. Tuck, AB, Elliott, BE, Hota, C,  et al. 2000Osteopontin-induced, integrin-dependent migration of human mammary epithelial cells involves activation of the hepatocyte growth factor receptor (Met).Clinical & Experimental Metastasis78465475Google Scholar
  11. Weinstat-Saslow, DL, Zabrenetzky, VS, VanHoutte, K, Frazier, WA, Roberts, DD, Steeg, PS 1994Transfection of thrombospondin 1 complementary DNA into a human breast carcinoma cell line reduces primary tumor growth, metastatic potential, and angiogenesis.Clinical & Experimental Metastasis5465046511Google Scholar
  12. Xuan, JW, Hota, C, Chambers, AF 1994Recombinant GST-human osteopontin fusion protein is functional in RGD-dependent cell adhesion.Clinical & Experimental Metastasis54247255Google Scholar
  13. Hortobagyi, GN 2001Adjuvant systemic therapy for early breast cancer: progress and controversies.Clinical & Experimental Metastasis718391842Google Scholar
  14. Cowley, S, Paterson, H, Kemp, P, Marshall, CJ 1994Activation of MAP kinase is necessary and sufficient for PC12 differentiation and for transformation of NIH 3T3 cells.Clinical & Experimental Metastasis77841852Google Scholar
  15. Traverse, S, Gomez, N, Paterson, H, Marshall, C, Cohen, P 1992Sustained activation of the mitogen-activated protein (MAP) kinase cascade may be required for differentiation of PC12 cells.Clinical & Experimental Metastasis288351355(Pt 2)Google Scholar
  16. Ray,  RM, Zimmerman,  BJ , McCormack , SA , Patel,  TB , Johnson,  LR 1999Polyamine depletion arrests cell cycle and induces inhibitors p21(Waf1/Cip1), p27(Kip1), and p53 in IEC-6 cellsClinical & Experimental Metastasis276C684C691Google Scholar
  17. Chen, Y, Alm, K, Vujcic, S, Kramer, DL, Kee, K, Diegelman, P,  et al. 2003The role of mitogen-activated protein kinase activation in determining cellular outcomes in polyamine analogue-treated human melanoma cells.Clinical & Experimental Metastasis6336193625Google Scholar
  18. Manni, A, Wechter, R, Verderame, MF, Mauger, D 1998Cooperativity between the polyamine pathway and HER-2neu in transformation of human mammary epithelial cells in culture: role of the MAPK pathway.Clinical & Experimental Metastasis76563570Google Scholar
  19. Hawighorst, T, Oura, H, Streit, M, Janes, L, Nguyen, L, Brown, LF,  et al. 2002Thrombospondin-1 selectively inhibits early-stage carcinogenesis and angiogenesis but not tumor lymphangiogenesis and lymphatic metastasis in transgenic mice.Clinical & Experimental Metastasis2179457956Google Scholar
  20. Gutierrez, LS, Suckow, M, Lawler, J, Ploplis, VA, Castellino, FJ 2003Thrombospondin 1–a regulator of adenoma growth and carcinoma progression in the APC(Min/+) mouse model.Clinical & Experimental Metastasis24199207Google Scholar
  21. Good, DJ, Polverini, PJ, Rastinejad, F, Beau , MM, Lemons, RS, Frazier, WA,  et al. 1990A tumor suppressor-dependent inhibitor of angiogenesis is immunologically and functionally indistinguishable from a fragment of thrombospondin.Clinical & Experimental Metastasis8766246628Google Scholar
  22. Jimenez, B, Volpert, OV, Crawford, SE, Febbraio, M, Silverstein, RL, Bouck, N 2000Signals leading to apoptosis-dependent inhibition of neovascularization by thrombospondin-1.Clinical & Experimental Metastasis64148Google Scholar
  23. Yamashita, Y, Kurohiji, T, Tuszynski, GP, Sakai, T, Shirakusa, T 1998Plasma thrombospondin levels in patients with colorectal carcinoma.Clinical & Experimental Metastasis82632638Google Scholar
  24. Urquidi, V, Sloan, D, Kawai, K, Agarwal, D, Woodman, AC, Tarin, D,  et al. 2002Contrasting expression of thrombospondin-1 and osteopontin correlates with absence or presence of metastatic phenotype in an isogenic model of spontaneous human breast cancer metastasis.Clinical & Experimental Metastasis86174Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Andrea Manni
    • 1
  • Sharlene Washington
    • 1
  • David Mauger
    • 2
  • Deborah A. Hackett
    • 1
  • Michael F. Verderame
    • 1
  1. 1.Departmnt of Medicine and Health Evaluation SciencesThe Pennsylvania State University College of MedicineHersheyUSA
  2. 2.Health Evaluation SciencesThe Pennsylvania State University College of MedicineHersheyUSA

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