Skip to main content
SpringerLink
Log in

Induction of heat shock protein 27 by hydroxyurea and its relationship to experimental metastasis

  • Published:
Clinical & Experimental Metastasis Aims and scope Submit manuscript

Abstract

Treatment of tumor cells with hydroxyurea (HU) has been shown to increase the experimental metastatic potential of these cells. We have previously described the induction of stress proteins (antioxidants) by in B16 murine melanoma cells and their relationship to the metastatic process. We have now investigated the induction by HU of another set of stress proteins, the heat shock proteins, and their role in experi-mental metastasis. HU markedly increased the cellular content of heat shock protein (hsp) 27 but not hsp 90, 72/73, or 60 as measured by immunoblotting. The induction of hsp27 protein was preceded specific increase in hsp27 mRNA. Furthermore, HU-treated cells were more thermotolerant. To investigate the functional role of hsp27, human hsp27 cDNA was constitutively overexpressed in B16 cells at seen in HU-treated cells. In separate experiments, we induced a global increase in hsps by heat shock. Neither the hsp27 transfectants nor the heat-shocked cells demonstrated an increase in their experimental metastatic capacity. We conclude that hsp27 protein is increased by HU by the specific induction of hsp27 mRNA in B16 melanoma cells but increased hsp27 protein is not responsible for the increase in experimental metastasis. Since high levels of hsp27 are associated with metastatic disease in breast and ovarian cancers, but not in our experimental system, the functional role of hsp27 in metastasis requires further study. © Rapid Science 1998

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Nicolson GL, 1991, Molecular mechanisms of cancer metastasis: tumor and host properties and the role of oncogenes and suppressor genes. Curr Opin Oncol, 3, 75-92.

    PubMed  Google Scholar 

  2. McMillan TJ, Rao J and Hart IR, 1986, Enhancement of experimental metastasis by pretreatment of tumour cells with hydroxyurea. Int J Cancer, 38, 61-5.

    PubMed  Google Scholar 

  3. McMillan TJ and Hart IR, 1986, Enhanced experimental metastatic capacity of a murine melanoma following pre-treatment with anticancer drugs. Clin Exp Metastasis, 4, 285-92.

    PubMed  Google Scholar 

  4. Eskenazi AE, Pinkas J, Whitin JC, Arguello F, Cohen HJ and Frantz CN, 1993, Role of antioxidant enzymes in the induction of increased experimental metastasis by hydroxyurea. J Natl Cancer Inst, 85, 711-21.

    PubMed  Google Scholar 

  5. Welch WJ, 1990, The mammalian stress response: cell physiology and biochemistry of stress proteins. In: Morimoto RI, Tissieres A and Georgopoulos C, eds. Stress Proteins in Biology and Medicine. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, pp. 223-78.

    Google Scholar 

  6. Morimoto RI, 1991, Heat shock: the role of transient inducible responses in cell damage, transformation, and differentiation. Cancer Cells, 3, 295-301.

    PubMed  Google Scholar 

  7. Ciocca DR, Oesterreich S, Chamness GC, McGuire WL and Fuqua SAW, 1993, Biological and clinical implications of heat shock protein 27000 (HSP27): a review. J Natl Cancer Inst, 85, 1558-70.

    PubMed  Google Scholar 

  8. Oesterreich S, Schunck H, Benndorf R and Bielka H, 1991, Cisplatin induces the small heat shock protein HSP25 and thermotolerance in Ehrlich ascites tumor cells. Biochem Biophys Res Commun, 180, 243-8.

    PubMed  Google Scholar 

  9. Moseley PL, York SJ and York J, 1989, Bleomycin induces the HSP-70 heat shock promoter in cultured cells. Am J Resp Cell Mol Biol, 1, 89-93.

    Google Scholar 

  10. Kroes RA, Abravaya K, Seidenfeld J and Morimoto RI, 1991, Selective activation of human heat shock gene transcription by nitrosourea antitumor drugs mediated by isocyanate-induced damage and activation of heat shock transcription factor. Proc Natl Acad Sci USA, 88, 4825-9.

    PubMed  Google Scholar 

  11. Li GC, 1987, Heat shock proteins: role in thermotolerance, drug resistance, and relationship to DNA topoisomerases. NCI Monogr, 4, 99-107.

    PubMed  Google Scholar 

  12. Oesterreich S, Weng C-N, Qiu M, Hilsenbeck SG, Osborne CK and Fuqua SAW, 1993, The small heat shock protein hsp27 is correlated with growth and drug resistance in human breast cancer cell lines. Cancer Res, 53, 4443-8.

    PubMed  Google Scholar 

  13. Huot J, Roy G, Lambert H, Chretien P and Landry J, 1991, Increased survival after treatments with anticancer agents of Chinese hamster cells expressing the human Mr 27,000 heat shock protein. Cancer Res, 51, 5245-52.

    PubMed  Google Scholar 

  14. Huot J, Houle F, Spitz DR and Landry J, 1996, HSP27 phosphorylation-mediated resistance against actin fragmentation and cell death induced by oxidative stress. Cancer Res, 56, 273-9.

    PubMed  Google Scholar 

  15. Stackpole CW, Alterman AL and Fornabaio DM, 1985, Growth characteristics of clonal cell populations constituting a B16 melanoma metastasis model system. Invasion Metastasis, 5, 125-43.

    PubMed  Google Scholar 

  16. Bradford MM, 1976, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt Biochem, 72, 248-54.

    Article  PubMed  Google Scholar 

  17. Laemmli UK, 1970, Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature, 227, 680.

    PubMed  Google Scholar 

  18. Badley JE, Bishop GA, St. John T and Frelinger J, 1988, A simple, rapid method for the purification of polyA+ RNA. BioTechniques, 6, 114-16.

    PubMed  Google Scholar 

  19. Feinberg AP and Vogelstein B, 1983, A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analyt Biochem, 132, 6-13.

    PubMed  Google Scholar 

  20. Hollander MC and Fornace AJ, 1990, Estimation of relative mRNA content by filter hybridization to a polythymidylate probe. Biotechniques, 9, 174-9.

    PubMed  Google Scholar 

  21. Miller AD and Rosman GJ, 1989, Improved retroviral vectors for gene transfer and expression. Biotechniques, 7, 980-90.

    PubMed  Google Scholar 

  22. Edwards DP, Adams DJ, Savage N and McGuire WL, 1980, Estrogen induced synthesis of specific proteins in human breast cancer cells. Biochem Biophys Res Commun, 93, 840-2.

    Google Scholar 

  23. Langdon SP, Rabiasz GJ, Hirst GL, et al. 1995, Expression of the heat shock protein HSP27 in human ovarian cancer. Clin Cancer Res, 1, 1603-9.

    PubMed  Google Scholar 

  24. Coccia DR, Puy LA and Fasoli LC, 1989, Study of estrogen receptor, progesterone receptor, and the estrogen-regulated Mr 24,000 protein in patients with carcinomas of the endometrium and cervix. Cancer Res, 49, 4298-304.

    PubMed  Google Scholar 

  25. Thor A, Benz C, Moore D, et al. 1991, Stress response protein (srp-27) determination in primary human breast carcinomas: clinical, histologic, and prognostic correlations. J Natl Cancer Inst, 83, 170-8.

    PubMed  Google Scholar 

  26. Tetu B, Lacass B, Bouchard H-L, Lagace R, Huot J and Landry J, 1992, Prognostic influence of hsp27 expression in malignant fibrous histiocytoma: a clinicopathological and immunohistochemical study. Cancer Res, 52, 2325-8.

    PubMed  Google Scholar 

  27. Ungar DR, Hailat N, Strahler JR, et al. 1994, Hsp27 expression neuroblastoma: correlation with disease stage. J Natl Cancer Inst, 86, 780-4.

    PubMed  Google Scholar 

  28. Strahler JR, Kuick R, Eckerskorn C, et al. 1990, Identification of two related markers for common acute lymphoblastic leukemia as heat shock proteins. J Clin Invest, 85, 200-7.

    PubMed  Google Scholar 

  29. Fuqua SA, Oesterreich S, Hilsenbeck SG, Von Hoff DD, Eckardt J and Osborne CK, 1994, Heat shock proteins and drug resistance. Breast Cancer Res Treat, 32, 67-71.

    Google Scholar 

  30. Schlappack OK, Zimmerman A and Hill RP, 1991, Glucose starvation and acidosis: effect on experimental metastatic potential, DNA content and MTX resistance of murine tumour cells. Br J Cancer, 64, 663-70.

    PubMed  Google Scholar 

  31. Young SD and Hill RP, 1990, Effects of reoxygenation on cells from hypoxic regions of solid tumors: anticancer drug sensitivity and metastatic potential. J Natl Cancer Inst, 82, 371-80.

    PubMed  Google Scholar 

  32. Spitz DR, Dewey WC and Li GC, 1987, Hydrogen peroxide or heat shock induces resistance to hydrogen peroxide in Chinese hamster fibroblasts. J Cell Physiol, 131, 364-73.

    Google Scholar 

  33. Tomasovic SP and Welch DR, 1986, Heat stress proteins and experimental cancer metastasis. Int J Hyperthermia, 2, 253-66.

    PubMed  Google Scholar 

  34. Muramatsu T, Kobayashi N, Tada H, Hatoko M and Shira, 1995, Induction of the 72-kD heat shock protein in human skin melanoma and squamous cell carcinoma cell lines. J Dermatol, 22, 907-15.

    PubMed  Google Scholar 

  35. Foss AJ, Alexander RA, Guille MJ, Hungerford JL, McCartney AC and Lightman S, 1995, Estrogen and progesterone receptor analysis in ocular melanoma. Ophthalmology, 102, 431-5.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Children's Cancer Foundation Research Laboratory, Department of Pediatrics and the Marlene and Stuart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA

    Allen E. Eskenazi, Jennifer Powers, Jan Pinkas & Christopher N. Frantz

  2. Division of Medical Oncology, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Steffi Oesterreich

Authors
  1. Allen E. Eskenazi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jennifer Powers
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jan Pinkas
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Steffi Oesterreich
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Suzanne A. W. Fuqua
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Christopher N. Frantz
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Eskenazi, A.E., Powers, J., Pinkas, J. et al. Induction of heat shock protein 27 by hydroxyurea and its relationship to experimental metastasis. Clin Exp Metastasis 16, 283–290 (1998). https://doi.org/10.1023/A:1006553127695

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006553127695

Search

Navigation