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Analysis of nuclear nestin localization in cell lines derived from neurogenic tumors

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Tumor Biology

Abstract

Nestin is a class VI intermediate filament protein expressed in the cytoplasm of stem and progenitor cells in the mammalian CNS during development. In adults, nestin is present only in a small subset of cells and tissues, including the subventricular zone of the adult mammalian brain, where neurogenesis occurs. Nestin expression has also been detected under such pathological conditions as ischemia, inflammation, and brain injury, as well as in various types of human solid tumors and their corresponding cell lines. Furthermore, nestin was recently found in the nuclei of glioblastoma, neuroblastoma, and angiosarcoma cells and it was proved to interact directly with the nuclear DNA in neuroblastoma cells. Here, we perform the first study of the intracellular distribution of nestin in cell lines derived from neurogenic tumors. Using immunodetection methods, we examined nestin expression in tumor-derived cell lines obtained from 11 patients with neuroblastoma, medulloblastoma, or glioblastoma multiforme. Besides its standard cytoplasmic localization, nestin was present in the nuclei of two neuroblastoma cell lines and one medulloblastoma cell line. Nestin was only present in the nuclei of cells with diffuse cytoplasmic staining for this protein, and the proportion of cells positive for nestin in nuclei, as well as the intensity of staining, varied. The presence of nestin in the nuclei was confirmed by both transmission electron microscopy and Western blotting. Our results indicate that the presence of nestin in the nuclei of tumor cells is not very rare, especially under in vitro conditions.

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References

  1. Dahlstrand J, Zimmerman LB, McKay RD, Lendahl U. Characterization of the human nestin gene reveals a close evolutionary relationship to neurofilaments. J Cell Sci. 1992;103:589–97.

    PubMed  CAS  Google Scholar 

  2. Lobo MV, Arenas MI, Alonso FJ, Gomez G, Bazan E, Paino CL, et al. Nestin, a neuroectodermal stem cell marker molecule, is expressed in Leydig cells of the human testis and in some specific cell types from human testicular tumours. Cell Tissue Res. 2004;316:369–76. doi:10.1007/s00441-003-0848-4.

    Article  PubMed  CAS  Google Scholar 

  3. Tamagno I, Schiffer D. Nestin expression in reactive astrocytes of human pathology. J Neurooncol. 2006;80:227–33. doi:10.1007/s11060-006-9181-6.

    Article  PubMed  CAS  Google Scholar 

  4. Faulkner JR, Herrmann JE, Woo MJ, Tamsey KE, Doan NB, Sofroniew NB. Reactive astrocytes protect tissue and preserve function after spinal cord injury. J Neurosci. 2004;24:2143–55. doi:10.1523/JNEUROSCI.3547-03.2004.

    Article  PubMed  CAS  Google Scholar 

  5. Mokry J, Pudil R, Ehrmann J, Cizkova D, Osterreicher J, Filip S, et al. Re-expression of nestin in the myocardium of postinfarcted patients. Virchows Arch. 2008;453:33–41. doi:10.1007/s00428-008-0631-8.

    Article  PubMed  CAS  Google Scholar 

  6. Krupkova Jr O, Loja T, Zambo I, Veselska R. Nestin expression in human tumors and tumor cell lines. Neoplasma. 2010;57:291–8. doi:10.4149/neo_2010_04_291.

    Article  PubMed  Google Scholar 

  7. Dahlstrand J, Collins VP, Lendahl U. Expression of the class VI intermediate filament nestin in human central nervous system tumors. Cancer Res. 1992;52:5334–41.

    PubMed  CAS  Google Scholar 

  8. Piras F, Perra MT, Murtas D, Minerba L, Floris C, Maxia C, et al. The stem cell marker nestin predicts poor prognosis in human melanoma. Oncol Rep. 2010;23:17–24. doi:10.3892/or_00000601.

    PubMed  Google Scholar 

  9. Sugawara K, Kurihara H, Negishi M, Saito N, Nakazato Y, Sasaki T, et al. Nestin as a marker for proliferative endothelium in gliomas. Lab Invest. 2002;82:345–51.

    PubMed  CAS  Google Scholar 

  10. Veselska R, Kuglik P, Cejpek P, Svachova H, Neradil J, Loja T, et al. Nestin expression in the cell lines derived from glioblastoma multiforme. BMC Cancer. 2006;6:32. doi:10.1186/1471-2407-6-32.

    Article  PubMed  Google Scholar 

  11. Sahlgren CM, Mikhailov A, Hellman J, Chou YH, Lendahl U, Goldman RD, et al. Mitotic reorganization of the intermediate filament protein nestin involves phosphorylation by cdc2 kinase. J Biol Chem. 2001;276:16456–63. doi:10.1074/jbc.M009669200.

    Article  PubMed  CAS  Google Scholar 

  12. Loja T, Chlapek P, Kuglik P, Pesakova M, Oltova A, Cejpek P, et al. Characterization of a GM7 glioblastoma cell line showing CD133 positivity and both cytoplasmic and nuclear localization of nestin. Oncol Rep. 2009;21:119–27. doi:10.3892/or_00000198.

    PubMed  CAS  Google Scholar 

  13. Thomas SK, Messam CA, Spengler BA, Biedler JL, Ross RA. Nestin is a potential mediator of malignancy in human neuroblastoma cells. J Biol Chem. 2004;279:27994–9. doi:10.1074/jbc.M312663200.

    Article  PubMed  CAS  Google Scholar 

  14. Yang X, Wu Q, Yu X, Xu CX, Ma BF, Zhang XM, et al. Nestin expression in different tumors and its relevance to malignant grade. J Clin Pathol. 2008;61:467–73. doi:10.1136/jcp.2007.047605.

    Article  PubMed  CAS  Google Scholar 

  15. Merigo F, Mucignat-Caretta C, Zancanaro C. Timing of neuronal intermediate filament proteins expression in the mouse vomeronasal organ during pre- and postnatal development. An immunohistochemical study. Chem Senses. 2005;30:707–17. doi:10.1093/chemse/bji063.

    Article  PubMed  CAS  Google Scholar 

  16. Loja T. Relationship between histopathological and molecular cytogenetic markers on chosen solid tumors in vitro. Dissertation: Masaryk University; 2008.

    Google Scholar 

  17. Ma YH, Mentlein R, Knerlich F, Kruse ML, Mehdorn HM, Held-Feindt J. Expression of stem cell markers in human astrocytomas of different WHO grades. J Neurooncol. 2008;86:31–45. doi:10.1007/s11060-007-9439-7.

    Article  PubMed  Google Scholar 

  18. Ehrmann J, Kolar Z, Mokry J. Nestin as a diagnostic and prognostic marker: immunohistochemical analysis of its expression in different tumours. J Clin Pathol. 2005;58:222–3. doi:10.1136/jcp.2004.021238.

    Article  PubMed  CAS  Google Scholar 

  19. Fusi A, Reichelt U, Busse A, Ochsenreither S, Rietz A, Maisel M, et al. Expression of the stem cell markers nestin and CD133 on circulating melanoma cells. J Invest Dermatol. 2010. doi:10.1038/jid.2010.285.

    PubMed  Google Scholar 

  20. Korja M, Finne J, Salmi TT, Kalimo H, Karikoski R, Tanner M, et al. Chromogenic in situ hybridization-detected hotspot MYCN amplification associates with Ki-67 expression and inversely with nestin expression in neuroblastomas. Mod Pathol. 2005;18:1599–605. doi:10.1038/modpathol.3800462.

    PubMed  CAS  Google Scholar 

  21. Sahlgren CM, Mikhailov A, Vaittinen S, Pallari HM, Kalimo H, Pant HC, et al. Cdk5 regulates the organization of nestin and its association with p35. Mol Cell Biol. 2003;23:5090–106. doi:10.1128/MCB.23.14.5090-5106.2003.

    Article  PubMed  CAS  Google Scholar 

  22. Sahlgren CM, Pallari HM, Chou YH, Goldman RD, Eriksson JE. A nestin scaffold links Cdk5/p35 signaling to oxidant-induced cell death. EMBOJ. 2006;25:4808–19. doi:10.1038/sj.emboj.7601366.

    Article  CAS  Google Scholar 

  23. Marvin MJ, Dahlstrand J, Lendahl U, McKay RD. A rod end deletion in the intermediate filament protein nestin alters its subcellular localization in neuroepithelial cells of transgenic mice. J Cell Sci. 1998;111:1951–61.

    PubMed  CAS  Google Scholar 

  24. Steinert PM, Chou YH, Prahlad V, Parry DA, Marekov LN, Wu KC, et al. A high molecular weight intermediate filament-associated protein in BHK-21 cells is nestin, a type VI intermediate filament protein. Limited co-assembly in vitro to form heteropolymers with type III vimentin and type IV alpha-internexin. J Biol Chem. 1999;274:9881–90.

    Article  PubMed  CAS  Google Scholar 

  25. Hartig R, Shoeman RL, Janetzko A, Tolstonog G, Traub P. DNA-mediated transport of the intermediate filament protein vimentin into the nucleus of cultured cells. J Cell Sci. 1998;111:3573–84.

    PubMed  CAS  Google Scholar 

  26. Traub P, Mothes E, Shoeman RL, Schroder R, Scherbarth A. Binding of nucleic acids to intermediate filaments of the vimentin type and their effects on filament formation and stability. J Biomol Struct Dyn. 1992;10:505–31.

    PubMed  CAS  Google Scholar 

  27. Luderus ME, de Graaf A, Mattia E, den Blaauwen JL, Grande MA, de Jong L, et al. Binding of matrix attachment regions to lamin B1. Cell. 1992;70:949–59.

    Article  PubMed  CAS  Google Scholar 

  28. Luderus ME, den Blaauwen JL, de Smit OJ, Compton DA, van Driel R. Binding of matrix attachment regions to lamin polymers involves single-stranded regions and the minor groove. Mol Cell Biol. 1994;14:6297–305.

    PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Dobromila Klemova and Johana Maresova for technical assistance. We also acknowledge Barbora Bujokova for confirming nestin expression in the SH-SY5Y cell line. This work was supported by Grant MUNI/A/0090/2009 of the Masaryk University as well as Grants IGA NR/9125-4 of the Ministry of Health of the Czech Republic and VZ MSM0021622415 of the Ministry of Education of the Czech Republic.

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Authors and Affiliations

  1. Laboratory of Tumor Biology and Genetics, Department of Experimental Biology, Faculty of Science, Masaryk University, Kotlarska 2, 61137, Brno, Czech Republic

    Olga Krupkova Jr, Tomas Loja, Martina Redova, Jakub Neradil & Renata Veselska

  2. Department of Pediatric Oncology, Masaryk University and University Hospital Brno, Cernopolni 9, 61300, Brno, Czech Republic

    Jakub Neradil, Karel Zitterbart, Jaroslav Sterba & Renata Veselska

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  1. Olga Krupkova Jr
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  2. Tomas Loja
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  3. Martina Redova
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  4. Jakub Neradil
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  7. Renata Veselska
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Correspondence to Renata Veselska.

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Krupkova, O., Loja, T., Redova, M. et al. Analysis of nuclear nestin localization in cell lines derived from neurogenic tumors. Tumor Biol. 32, 631–639 (2011). https://doi.org/10.1007/s13277-011-0162-9

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  • DOI: https://doi.org/10.1007/s13277-011-0162-9

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