BACKGROUND: Nestin was originally identified as a class VI intermediate filament protein that is expressed in stem cells and progenitor cells in the mammalian CNS during development. This protein is replaced in the adult organism by other intermediate filament proteins; however, nestin may be re-expressed under certain pathological conditions such as ischemia, inflammation, brain injury, and neoplastic transformation. Nestin has been detected in many kinds of tumors, especially in tumors derived from the CNS. Co-expression of nestin and the CD133 surface molecule is considered to be a marker for cancer stem cells in neurogenic tumors. Our work was aimed at a detailed study of nestin expression in osteosarcomas and osteosarcoma-derived cell lines. METHODS: Using immunodetection methods, we examined nestin in tumor tissue samples from 18 patients with osteosarcomas. We also successfully established permanent cell lines from the tumor tissue of 4 patients and immunodetection of nestin and CD133 was performed on these cell lines. RESULTS: Nestin-positive tumor cells were immunohistochemically detected in all of the examined osteosarcomas, but the proportion of these cells that were positively stained as well as the intensity of staining varied. Nestin-positive cells were rarely observed in 2 tumor samples, and the remaining 16 tumor samples showed various nestin expression patterns ranging from very sporadic occurrence to an overwhelming proportion of cells with strong positive staining. Three of the established osteosarcoma cell lines were demonstrated to be nestin-positive, and only one cell line showed no expression of nestin; this finding corresponds with the rare occurrence of nestin-positive cells in the respective tumor sample. Moreover, three of these osteosarcoma cell lines were undoubtedly proven to be Nes+/CD133+. CONCLUSION: Our results represent the first evidence of nestin expression in osteosarcomas and suggest the possible occurrence of cells with a stem-like phenotype in these tumors.

Laboratory of Tumor Biology and Genetics Institute of Experimental Biology School of Science Masaryk University Kotlarska 2 611 37 Brno Czech Republic

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Hockfield S, McKay RD. Identification of major cell classes in the developing mammalian nervous system. J Neurosci. 1985;5:3310–3328. PubMed PMC

Lendahl U, Zimmerman LB, McKay RDG. CNS stem cells express a new class of intermediate filament protein. Cell. 1990;60:585–595. doi: 10.1016/0092-8674(90)90662-X. PubMed DOI

Dahlstrand J, Zimmerman LB, McKay RDG, Lendahl U. Characterization of the human nestin gene reveals a close evolutionary relationship to neurofilaments. J Cell Sci. 1992;103:589–597. PubMed

Zimmerman L, Parr B, Lendahl U, Cunningham M, McKay R, Gavin B, Mann J, Vassileva G, McMahon A. Independent regulatory elements in the nestin gene direct transgene expression to neural stem cells or muscle precursors. Neuron. 1994;12:11–24. doi: 10.1016/0896-6273(94)90148-1. PubMed DOI

Dahlstrand J, Collins VP, Lendahl U. Expression of the class VI intermediate filament nestin in human central nervous system tumors. Cancer Res. 1992;52:5334–5341. PubMed

Dahlstrand J, Lardelli M, Lendahl U. Nestin mRNA expression correlates with the central nervous system progenitor cell in many, but not all, regions of developing central nervous system. Brain Res Dev Brain Res. 1995;84:109–129. doi: 10.1016/0165-3806(94)00162-S. PubMed DOI

Morshead CM, Reynolds BA, Craig CG, McBurney MW, Staines WA, Morassutti D, Weiss S, van der Kooy D. Neural stem cells in the adult mammalian forebrain: A relatively quiescent subpopulation of subependymal cells. Neuron. 1994;13:1071–1082. doi: 10.1016/0896-6273(94)90046-9. PubMed DOI

Andressen C, Stöcker E, Klinz FJ, Lenka N, Hescheler J, Fleischmann B, Arnhold S, Addicks K. Nestin-specific green fluorescent protein expression in embryonic stem-cell neural precursor cells used for transplantation. Stem Cells. 2001;19:419–424. doi: 10.1634/stemcells.19-5-419. PubMed DOI

Krum JM, Rosenstein JM. Transient coexpression of nestin, GFAP, and vascular endothelial growth factor in mature reactive astroglial following neural grafting or brain wounds. Exp Neurol. 1999;160:348–360. doi: 10.1006/exnr.1999.7222. PubMed DOI

Ha Y, Choi JU, Yoon DH, Cho YE, Kim TS. Nestin and small heat shock protein expression on reactive astrocytes and endothelial cells in cerebral abscess. Neurosci Res. 2002;44:207–212. doi: 10.1016/S0168-0102(02)00126-8. PubMed DOI

Smits A, van Grieken D, Hartman M, Lendahl U, Funa K, Nistér M. Coexpression of platelet-derived growth factor alpha and beta receptors on medulloblastomas and other primitive neuroectodermal tumors is consistent with an immature stem cell and neuronal derivation. Lab Invest. 1996;74:188–198. PubMed

Khoddami M, Becker LE. Immunohistochemistry of medulloepithelioma and neural tube. Pediatr Pathol Lab Med. 1997;17:913–925. doi: 10.1080/107710497174363. PubMed DOI

Brisson C, Lelong-Rebel I, Mottolese C, Jouvet A, Fèvre-Montange M, Saint Pierre G, Rebel G, Belin MF. Establishment of human tumoral ependymal cell lines and coculture with tubular-like human endothelial cells. Int J Oncol. 2002;21:775–785. PubMed

Almqvist PM, Mah R, Lendahl U, Jacobsson B, Hendson G. Immunohistochemical detection of nestin in pediatric brain tumors. J Histochem Cytochem. 2002;50:147–158. PubMed

Duggal N, Hammond RR. Nestin expression in ganglioglioma. Exp Neurol. 2002;174:89–95. doi: 10.1006/exnr.2001.7838. PubMed DOI

Donner LR, Teshima I. Peripheral medulloepithelioma: an immunohistochemical, ultrastructural, and cytogenetic study of a rare, chemotherapy-sensitive, pediatric tumor. Am J Surg Pathol. 2003;27:1008–1012. doi: 10.1097/00000478-200307000-00018. PubMed DOI

Kim BJ, Kim SS, Kim YI, Paek SH, Lee YD, Suh-Kim H. Forskolin promotes astroglial differentiation of human central neurocytoma cells. Exp Mol Med. 2004;36:52–56. PubMed

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;58:222–223. doi: 10.1136/jcp.2004.021238. PubMed DOI PMC

You H, Kim YI, Im SY, Suh-Kim H, Paek SH, Park SH, Kim DG, Jung HW. Immunohistochemical study of central neurocytoma, subependymoma, and subependymal giant cell astrocytoma. J Neurooncol. 2005;74:1–8. doi: 10.1007/s11060-004-2354-2. PubMed DOI

Fevre-Montange M, Hasselblatt M, Figarella-Branger D, Chauveinc L, Champier J, Saint-Pierre G, Taillandier L, Coulon A, Paulus W, Fauchon F, Jouvet A. Prognosis and histopathologic features in papillary tumors of the pineal region: a retrospective multicenter study of 31 cases. J Neuropathol Exp Neurol. 2006;65:1004–1011. PubMed

Ikota H, Kinjo S, Yokoo H, Nakazato Y. Systematic immunohistochemical profiling of 378 brain tumors with 37 antibodies using tissue microarray technology. Acta Neuropathol. 2006;111:475–482. doi: 10.1007/s00401-006-0060-1. PubMed DOI

Rani SB, Mahadevan A, Anilkumar SR, Raju TR, Shankar SK. Expression of nestin – a stem cell associated intermediate filament in human CNS tumours. Indian J Med Res. 2006;124:269–280. PubMed

Schiffer D, Manazza A, Tamagno I. Nestin expression in neuroepithelial tumors. Neurosci Lett. 2006;400:80–85. doi: 10.1016/j.neulet.2006.02.034. PubMed DOI

Zhang QB, Ji XY, Huang Q, Dong J, Zhu YD, Lan Q. Differentiation profile of brain tumor stem cells: a comparative study with neural stem cells. Cell Res. 2006;16:909–915. doi: 10.1038/sj.cr.7310104. PubMed DOI

Kobayashi M, Sjöberg G, Söderhäll S, Lendahl U, Sandstedt B, Sejersen T. Pediatric rhabdomyosarcomas express the intermediate filament nestin. Pediatr Res. 1998;43:386–392. doi: 10.1203/00006450-199803000-00013. PubMed DOI

Tsujimura T, Makiishi-Shimobayashi C, Lundkvist J, Lendahl U, Nakasho K, Sugihara A, Iwasaki T, Mano M, Yamada N, Yamashita K, Toyosaka A, Terada N. Expression of the intermediate filament nestin in gastrointestinal stromal tumors and interstitial cells of Cajal. Am J Pathol. 2001;158:817–823. PubMed PMC

Sarlomo-Rikala M, Tsujimura T, Lendahl U, Miettinen M. Patterns of nestin and other intermediate filament expression distinguish between gastrointestinal stromal tumors, leiomyomas and schwannomas. APMIS. 2002;110:499–507. doi: 10.1034/j.1600-0463.2002.100608.x. PubMed DOI

Hou YY, Tan YS, Xu JF, Wang XN, Lu SH, Ji Y, Wang J, Zhu XZ. Schwannoma of the gastrointestinal tract: a clinicopathological, immunohistochemical and ultrastructural study of 33 cases. Histopathology. 2006;48:536–545. doi: 10.1111/j.1365-2559.2006.02370.x. PubMed DOI

Parfitt JR, McLean CA, Joseph MG, Streutker CJ, Al-Haddad S, Driman DK. Granular cell tumours of the gastrointestinal tract: expression of nestin and clinicopathological evaluation of 11 patients. Histopathology. 2006;48:424–430. doi: 10.1111/j.1365-2559.2006.02352.x. PubMed DOI

Florenes VA, Holm R, Myklebost O, Lendahl U, Fodstad O. Expression of the neuroectodermal intermediate filament nestin in human melanomas. Cancer Res. 1994;54:354–356. PubMed

Klein WM, Wu BP, Zhao S, Wu H, Klein-Szanto AJ, Tahan SR. Increased expression of stem cell markers in malignant melanoma. Mod Pathol. 2007;20:102–107. doi: 10.1038/modpathol.3800720. PubMed DOI

Aihara M, Sugawara K, Torii S, Hosaka M, Kurihara H, Saito N, Takeuchi T. Angiogenic endothelium-specific nestin expression is enhanced by the first intron of the nestin gene. Lab Invest. 2004;84:1581–1592. doi: 10.1038/labinvest.3700186. PubMed DOI

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;279:27994–27999. doi: 10.1074/jbc.M312663200. PubMed DOI

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

Pilkington GJ. Cancer stem cells in the mammalian central nervous system. Cell Prolif. 2005;38:423–433. doi: 10.1111/j.1365-2184.2005.00358.x. PubMed DOI PMC

Liu G, Yuan X, Zeng Z, Tunici P, Ng H, Abdulkadir IR, Lu L, Irvin D, Black KL, Yu JS. Analysis of gene expression and chemoresistance of CD133+ cancer stem cells in glioblastoma. Mol Cancer. 2006;5:67. doi: 10.1186/1476-4598-5-67. PubMed DOI PMC

Yang XH, Wu LQ, Yu XB, Xu CX, Ma BF, Zhang XM, Li SN, Lahn BT, Xiang AP. Nestin expression in different tumors and its relevance to malignant grade. J Clin Pathol. 2007;61:467–473. doi: 10.1136/jcp.2007.047605. PubMed DOI

Somers GR, Zielenska M, Abdullah S, Sherman C, Chan S, Thorner PS. Expression of MYCN in pediatric synovial sarcoma. Mod Pathol. 2007;20:734–741. doi: 10.1038/modpathol.3800792. PubMed DOI

Olsen SH, Thomas DG, Lucas DR. Cluster analysis of immunohistochemical profiles in synovial sarcoma, malignant peripheral nerve sheath tumor, and Ewing sarcoma. Mod Pathol. 2006;19:659–668. doi: 10.1038/modpathol.3800569. PubMed DOI

Olempska M, Eisenach PA, Ammerpohl O, Ungefroren H, Fandrich F, Kalthoff H. Detection of tumor stem cell markers in pancreatic carcinoma cell lines. Hepatobiliary Pancreat Dis Int. 2007;6:92–97. PubMed

Jordan CT. Cancer stem cell biology: from leukemia to solid tumors. Curr Opin Cell Biol. 2004;16:708–712. doi: 10.1016/j.ceb.2004.09.002. PubMed DOI

Burkert J, Wright NA, Alison MR. Stem cells and cancer: an intimate relationship. J Pathol. 2006;209:287–297. doi: 10.1002/path.2016. PubMed DOI

Li F, Tiede B, Massagué J, Kang Y. Beyond tumorigenesis: cancer stem cells in metastasis. Cell Res. 2007;17:3–14. doi: 10.1038/sj.cr.7310118. PubMed DOI

Vaish M. Mismatch repair deficiencies transforming stem cells into cancer stem cells and therapeutic implications. Mol Cancer. 2007;6:26. doi: 10.1186/1476-4598-6-26. PubMed DOI PMC

Neuzil J, Stantic M, Zobalova R, Chladova J, Wang X, Prochazka L, Dong L, Andera L, Ralph SJ. Tumour-initiating cells vs. cancer 'stem' cells and CD133: What's in the name? Biochem Biophys Res Commun. 2007;355:855–859. doi: 10.1016/j.bbrc.2007.01.159. PubMed DOI

Ratajczak M. Cancer stem cells – normal stem cells "Jedi" that went over to the "dark side". Folia Histochem Cytobiol. 2005;43:175–181. PubMed

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