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Inducible Expression of p57KIP2 Inhibits Glioma Cell Motility and Invasion

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Abstract

To examine the role of p57KIP2 in human malignant glioma cells, we studied its expression in a panel of human malignant glioma specimens by western blot and immunohistochemical analysis. To determine the effects of p57KIP2 expression on the phenotype of glioma cells, we analyzed two inducible stably transfected p57KIP2 expressing glioma cell lines. Expression of p57KIP2 was induced in U373 and U87 malignant gliomacells with doxycycline using the tetracycline repressor system. A phagokinetic track assay on gold particles was used to investigate differences in cell migration between p57KIP2 expressing and non-expressing control cells. The effects of the extracellular matrix (ECM) on U373 motility was determined in p57+ and p57− cells on surfaces coated with 5 μg/cm2 of fibronectin, laminin, type I and type IV collagens. The invasion of p57+ and p57− glioma cells across BD Biocoat Matrigel invasion chambers was then determined. p57KIP2 was weakly expressed in 4/6 glioblastoma (GBM) specimens by western blot. By immunohistochemistry, p57KIP2 immunoreactivity was positive in 8/40 GBMs, and was primarily nuclear in location. The motility of U373 glioma cells was significantly reduced after p57KIP2 induction. The presence of ECM proteins did not further alter the motility of p57+ and p57− glioma cells. The results of the invasion chamber assay showed that p57+ cells exhibited a 35% reduction in their invasive capacity as compared to p57− cells. These data suggest that p57KIP2 is expressed in at least some malignant gliomas. Inducible expression of 57KIP2 in cell lines deficient in this cyclin-dependent kinase inhibitor reduces their motility and invasiveness.

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References

  1. Holland EC: Glioblastoma multiforme: the terminator. Proc Natl Acad Sci USA 97: 6242–6244, 2000

    Article  PubMed  CAS  Google Scholar 

  2. Louis DN, Holland EC, Cairncross JG: Glioma classification: a molecular reappraisal. Am J Pathol 159: 779–786, 2001

    PubMed  CAS  Google Scholar 

  3. Dirks PB, Hubbard SL, Murakami M, Rutka JT: Cyclin and cyclin-dependent kinase expression in human astrocytoma cell lines. J Neuropathol Exp Neurol 56: 291–300, 1997

    PubMed  CAS  Google Scholar 

  4. Dirks PB, Rutka JT: Current concepts in neuro-oncology: the cell cycle — a review. Neurosurgery 40: 1000–1013, 1997 (discussion 1013–1005)

    Article  PubMed  CAS  Google Scholar 

  5. Tsugu A, Sakai K, Dirks PB, Jung S, Weksberg R, Fei YL, Mondal S, Ivanchuk S, Ackerley C, Hamel PA et al.: Expression of p57(KIP2) potently blocks the growth of human astrocytomas and induces cell senescence. Am J Pathol 157: 919–932, 2000

    PubMed  CAS  Google Scholar 

  6. Mainprize TG, Taylor MD, Rutka JT, Dirks PB: Cip/Kip cell-cycle inhibitors: a neuro-oncological perspective. J Neurooncol 51: 205–218, 2001

    Article  PubMed  CAS  Google Scholar 

  7. Matsuoka S, Edwards MC, Bai C, Parker S, Zhang P, Baldini A, Harper JW, Elledge SJ: p57KIP2, a structurally distinct member of the p21CIP1 Cdk inhibitor family, is a candidate tumor suppressor gene. Genes Dev 9: 650–662, 1995

    PubMed  CAS  Google Scholar 

  8. Giese A, Loo MA, Rief MD, Tran N, Berens ME: Substrates for astrocytoma invasion. Neurosurgery 37: 294–301, 1995 (discussion 301–292)

    PubMed  CAS  Google Scholar 

  9. Yoshizato K, Zapf S, Westphal M, Berens ME, Giese A: Thromboxane synthase inhibitors induce apoptosis in migration-arrested glioma cells. Neurosurgery 50: 343–354, 2002

    Article  PubMed  Google Scholar 

  10. Westermark B, Ponten J, Hugosson R: Determinants for the establishment of permanent tissue culture lines from human gliomas. Acta Pathol Microbiol Scand [A] 81: 791–805, 1973

    CAS  Google Scholar 

  11. Baron U, Gossen M, Bujard H: Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential. Nucleic Acids Res 25, 2723–2729, 1997

    Article  PubMed  CAS  Google Scholar 

  12. Gossen M, Bujard H: Studying gene function in eukaryotes by conditional gene inactivation. Annu Rev Genet 36: 153–173, 2002

    Article  PubMed  CAS  Google Scholar 

  13. Gossen M, Bujard H: Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci USA 89: 5547–5551, 1992

    Article  PubMed  CAS  Google Scholar 

  14. Gossen M, Bujard H: Efficacy of tetracycline-controlled gene expression is influenced by cell type: commentary. Biotechniques 19: 213–216, 1995. (Discussion 216–217)

    PubMed  CAS  Google Scholar 

  15. Gossen M, Freundlieb S, Bender G, Muller G, Hillen W, Bujard H: Transcriptional activation by tetracyclines in mammalian cells. Science 268, 1766–1769, 1995

    PubMed  CAS  Google Scholar 

  16. Resnitzky D, Gossen M, Bujard H, Reed SI: Acceleration of the G1/S phase transition by expression of cyclins D1 and E with an inducible system. Mol Cell Biol 14: 1669–1679, 1994

    PubMed  CAS  Google Scholar 

  17. Albrecht-Buehler G: The phagokinetic tracks of 3T3 cells. Cell 11: 395–404, 1977

    Article  PubMed  CAS  Google Scholar 

  18. Albrecht-Buehler G: Phagokinetic tracks of 3T3 cells: parallels between the orientation of track segments and of cellular structures which contain actin or tubulin. Cell 12: 333–339, 1977

    Article  PubMed  CAS  Google Scholar 

  19. Albrecht-Buehler G: The tracks of moving cells. Sci Am 238, 68–76, 1978

    Article  PubMed  CAS  Google Scholar 

  20. Albini A, Iwamoto Y, Kleinman HK, Martin GR, Aaronson SA, Kozlowski JM, McEwan RN: A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res 47: 3239–3245, 1987

    PubMed  CAS  Google Scholar 

  21. Giese A, Loo MA, Tran N, Haskett D, Coons SW, Berens ME: Dichotomy of astrocytoma migration and proliferation. Int J Cancer 67: 275–282, 1996

    Article  PubMed  CAS  Google Scholar 

  22. Mariani L, Beaudry C, McDonough WS, Hoelzinger DB, Demuth T, Ross KR, Berens T, Coons SW, Watts G, Trent JM et al.: Glioma cell motility is associated with reduced transcription of proapoptotic and proliferation genes: a cDNA microarray analysis. J Neurooncol 53: 161–176, 2001

    Article  PubMed  CAS  Google Scholar 

  23. Mariani L, Beaudry C, McDonough WS, Hoelzinger DB, Kaczmarek E, Ponce F, Coons SW, Giese A, Seiler RW, Berens ME: Death-associated protein 3 (Dap-3) is over-expressed in invasive glioblastoma cells in vivo and in glioma cell lines with induced motility phenotype in vitro. Clin Cancer Res 7: 2480–2489, 2001

    PubMed  CAS  Google Scholar 

  24. Ito Y, Takeda T, Sakon M, Tsujimoto M, Monden M, Matsuura N: Expression of p57/Kip2 protein in hepatocellular carcinoma. Oncology 61: 221–225, 2001

    Article  PubMed  CAS  Google Scholar 

  25. Nakai S, Masaki T, Shiratori Y, Ohgi T, Morishita A, Kurokohchi K, Watanabe S, Kuriyama S: Expression of p57(KIP2) in hepatocellular carcinoma: relationship between tumor differentiation and patient survival. Int J Oncol 20: 769–775, 2002

    PubMed  CAS  Google Scholar 

  26. Rosenberg E, Demopoulos RI, Zeleniuch-Jacquotte A, Yee H, Sorich J, Speyer JL, Newcomb EW: Expression of cell cycle regulators p57(KIP2), cyclin D1, cyclin E in epithelial ovarian tumors and survival. Hum Pathol 32: 808–813, 2001

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Neurosurgery, Shinshu University School of Medicine, Japan

    K Sakai, K Hongo & S Kobayashi

  2. Department of Organ Regeneration, Institutes of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University School of Medicine, Japan

    J Nakayama

  3. Department of Neurosurgery, Tokai University School of Medicine, Japan

    A Tsugu

  4. The Arthur and Sonia Labatt Brain Tumour Research Centre, The Hospital for Sick Children, and the Division of Neurosurgery, University of Toronto, Canada

    A Peraud, T Mainprize & James T. Rutka

Authors
  1. K Sakai
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  2. A Peraud
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  3. T Mainprize
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  4. J Nakayama
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  5. A Tsugu
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  6. K Hongo
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  7. S Kobayashi
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  8. James T. Rutka
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Sakai, K., Peraud, A., Mainprize, T. et al. Inducible Expression of p57KIP2 Inhibits Glioma Cell Motility and Invasion. J Neurooncol 68, 217–223 (2004). https://doi.org/10.1023/B:NEON.0000033380.08940.c8

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  • DOI: https://doi.org/10.1023/B:NEON.0000033380.08940.c8

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