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Detection of primary cilia in human glioblastoma

Abstract

Glioblastoma (GBM) is the most common malignant adult brain tumor and carries a poor prognosis due to primary and acquired resistance. While many cellular features of GBM have been documented, it is unclear if cells within these tumors extend a primary cilium, an organelle whose associated signaling pathways may regulate proliferation, migration, and survival of neural precursor and tumor cells. Using immunohistochemical and electron microscopy (EM) techniques, we screened human GBM tumor biopsies and primary cell lines for cilia. Immunocytochemical staining of five primary GBM cell lines revealed that between 8 and 25 % of the cells in each line possessed gamma tubulin-positive basal bodies from which extended acetylated, alpha-tubulin-positive axonemes. EM analyses confirmed the presence of cilia at the cell surface and revealed that their axonemes contained organized networks of microtubules, a structural feature consistent with our detection of IFT88 and Arl13b, two trafficked cilia proteins, along the lengths of the axonemes. Notably, cilia were detected in each of 23 tumor biopsies (22 primary and 1 recurrent) examined. These cilia were distributed across the tumor landscape including regions proximal to the vasculature and within necrotic areas. Moreover, ciliated cells within these tumors co-stained with Ki67, a marker for actively dividing cells, and ZEB1, a transcription factor that is upregulated in GBM and linked to tumor initiation, invasion, and chemoresistance. Collectively, our data show that subpopulations of cells within human GBM tumors are ciliated. In view of mounting evidence supporting roles of primary cilia in tumor initiation and propagation, it is likely that further study of the effects of cilia on GBM tumor cell function will improve our understanding of GBM pathogenesis and may provide new directions for GBM treatment strategies.

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Acknowledgments

We would like to thank the UF COM Core EM Facility and R. Martuscello for technical assistance, and T. Caspary for the rabbit Arl13b antibody. We would also like to thank B. Frentzen and R. McTiernan of the Florida Center for Brain Tumor Research for providing us with tumor samples. This work was supported, in part, by funds from the McKnight Brain Research Foundation and the Evelyn F. and William L. McKnight Brain Institute at the University of Florida (to M.R.S., B.A.S and D.A.S.), an American Cancer Society Chris DiMarco Institutional Research Grant Junior Investigator Award (to M.R.S), and an American Cancer Society Research Scholar Grant (#RSG-13-031-01-DDC) (to M.R.S.). M.R.S. would like to dedicate this study to the memory of his younger brother, Andrew T. Sarkisian, who died in 2007 at the age of 28 from blood cancer.

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The authors declare that they have no conflict of interest.

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Correspondence to Matthew R. Sarkisian.

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Sarkisian, M.R., Siebzehnrubl, D., Hoang-Minh, L. et al. Detection of primary cilia in human glioblastoma. J Neurooncol 117, 15–24 (2014). https://doi.org/10.1007/s11060-013-1340-y

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Keywords

  • Glioma
  • Cilium
  • Intraflagellar transport
  • Cancer