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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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.
Conflicts of interest
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.PubMedGoogle Scholar
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.PubMedCrossRefGoogle Scholar
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.PubMedCrossRefGoogle Scholar
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.PubMedGoogle Scholar
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.PubMedCrossRefGoogle Scholar
Loja T. Relationship between histopathological and molecular cytogenetic markers on chosen solid tumors in vitro. Dissertation: Masaryk University; 2008.Google Scholar
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.PubMedGoogle Scholar
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.PubMedGoogle Scholar
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.PubMedGoogle Scholar
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.PubMedCrossRefGoogle Scholar
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.PubMedGoogle Scholar
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.PubMedGoogle Scholar
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.PubMedCrossRefGoogle Scholar
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.PubMedGoogle Scholar