SOX3 can promote the malignant behavior of glioblastoma cells
- 272 Downloads
Glioblastoma is the most common and lethal adult brain tumor. Despite current therapeutic strategies, including surgery, radiation and chemotherapy, the median survival of glioblastoma patients is 15 months. The development of this tumor depends on a sub-population of glioblastoma stem cells governing tumor propagation and therapy resistance. SOX3 plays a role in both normal neural development and carcinogenesis. However, little is known about its role in glioblastoma. Thus, the aim of this work was to elucidate the role of SOX3 in glioblastoma.
SOX3 expression was assessed using real-time quantitative PCR (RT-qPCR), Western blotting and immunohistochemistry. MTT, immunocytochemistry and Transwell assays were used to evaluate the effects of exogenous SOX3 overexpression on the viability, proliferation, migration and invasion of glioblastoma cells, respectively. The expression of Hedgehog signaling pathway components and autophagy markers was assessed using RT-qPCR and Western blot analyses, respectively.
Higher levels of SOX3 expression were detected in a subset of primary glioblastoma samples compared to those in non-tumoral brain tissues. Exogenous overexpression of this gene was found to increase the proliferation, viability, migration and invasion of glioblastoma cells. We also found that SOX3 up-regulation was accompanied by an enhanced activity of the Hedgehog signaling pathway and by suppression of autophagy in glioblastoma cells. Additionally, we found that SOX3 expression was elevated in patient-derived glioblastoma stem cells, as well as in oncospheres derived from glioblastoma cell lines, compared to their differentiated counterparts, implying that SOX3 expression is associated with the undifferentiated state of glioblastoma cells.
From our data we conclude that SOX3 can promote the malignant behavior of glioblastoma cells.
KeywordsSOX3 Glioblastoma Glioblastoma stem cells Migration Hedgehog signaling Autophagy
This work was supported by the Ministry of Education, Science and Technological Development, Republic of Serbia (Grant No: 173051) and by the Serbian Academy of Sciences and Arts (Grant No: F 24). Jelena Marjanovic Vicentic received a grant from the IBRO-InEurope Short Stay Grants Program.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
The study was approved by the Ethics Committee of the Biodonostia Institute and Hospital Donostia and by the Ethics Committee of the Institute of Molecular Genetics and Genetic Engineering, University of Belgrade. It was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.
Informed consent was obtained from all participants included in the study.
- 3.R. Stupp, W. P. Mason, M. J. van den Bent, M. Weller, B. Fisher, M. J. Taphoorn, K. Belanger, A. A. Brandes, C. Marosi, U. Bogdahn, J. Curschmann, R. C. Janzer, S. K. Ludwin, T. Gorlia, A. Allgeier, D. Lacombe, J. G. Cairncross, E. Eisenhauer, R. O. Mirimanoff, R. for the European Organisation for Research and Treatment of Cancer Brain Tumor and Radiotherapy Groups and the National Cancer Institute of Canada Clinical Trials Group, Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N. Engl. J. Med. 352, 987-996 (2005)Google Scholar
- 5.C.W. Brennan, R.G. Verhaak, A. McKenna, B. Campos, H. Noushmehr, S.R. Salama, S. Zheng, D. Chakravarty, J.Z. Sanborn, S.H. Berman, R. Beroukhim, B. Bernard, C.J. Wu, G. Genovese, I. Shmulevich, J. Barnholtz-Sloan, L. Zou, R. Vegesna, S.A. Shukla, G. Ciriello, W.K. Yung, W. Zhang, C. Sougnez, T. Mikkelsen, K. Aldape, D.D. Bigner, E.G. Van Meir, M. Prados, A. Sloan, K.L. Black, J. Eschbacher, G. Finocchiaro, W. Friedman, D.W. Andrews, A. Guha, M. Iacocca, B.P. O'Neill, G. Foltz, J. Myers, D.J. Weisenberger, R. Penny, R. Kucherlapati, C.M. Perou, D.N. Hayes, R. Gibbs, M. Marra, G.B. Mills, E. Lander, P. Spellman, R. Wilson, C. Sander, J. Weinstein, M. Meyerson, S. Gabriel, P.W. Laird, D. Haussler, G. Getz, L. Chin, T.R. Network, The somatic genomic landscape of glioblastoma. Cell 155, 462–477 (2013)CrossRefPubMedPubMedCentralGoogle Scholar
- 7.M.L. Suva, E. Rheinbay, S.M. Gillespie, A.P. Patel, H. Wakimoto, S.D. Rabkin, N. Riggi, A.S. Chi, D.P. Cahill, B.V. Nahed, W.T. Curry, R.L. Martuza, M.N. Rivera, N. Rossetti, S. Kasif, S. Beik, S. Kadri, I. Tirosh, I. Wortman, A.K. Shalek, O. Rozenblatt-Rosen, A. Regev, D.N. Louis, B.E. Bernstein, Reconstructing and reprogramming the tumor-propagating potential of glioblastoma stem-like cells. Cell 157, 580–594 (2014)CrossRefPubMedPubMedCentralGoogle Scholar
- 12.J. Holmberg, X. He, I. Peredo, A. Orrego, G. Hesselager, C. Ericsson, O. Hovatta, S.M. Oba-Shinjo, S.K. Marie, M. Nister, J. Muhr, Activation of neural and pluripotent stem cell signatures correlates with increased malignancy in human glioma. PLoS One 6, e18454 (2011)CrossRefPubMedPubMedCentralGoogle Scholar
- 13.A.D. Berezovsky, L.M. Poisson, D. Cherba, C.P. Webb, A.D. Transou, N.W. Lemke, X. Hong, L.A. Hasselbach, S.M. Irtenkauf, T. Mikkelsen, A.C. deCarvalho, Sox2 promotes malignancy in glioblastoma by regulating plasticity and astrocytic differentiation. Neoplasia 16, 193–206, 206 e119–125 (2014)CrossRefPubMedPubMedCentralGoogle Scholar
- 14.I. Garcia, J. Aldaregia, J. Marjanovic Vicentic, P. Aldaz, L. Moreno-Cugnon, S. Torres-Bayona, E. Carrasco-Garcia, L. Garros-Regulez, L. Egana, A. Rubio, S. Pollard, M. Stevanovic, N. Sampron, A. Matheu, Oncogenic activity of SOX1 in glioblastoma. Sci. Rep. 7, 46575 (2017)CrossRefPubMedPubMedCentralGoogle Scholar
- 16.K.S. Alatzoglou, A. Azriyanti, N. Rogers, F. Ryan, N. Curry, C. Noakes, P. Bignell, G.W. Hall, A.S. Littooij, D. Saunders, P. Thomas, H. Stewart, M.T. Dattani, SOX3 deletion in mouse and human is associated with persistence of the craniopharyngeal canal. J. Clin. Endocrinol. Metab. 99, E2702–E2708 (2014)CrossRefPubMedGoogle Scholar
- 26.S.M. Pollard, K. Yoshikawa, I.D. Clarke, D. Danovi, S. Stricker, R. Russell, J. Bayani, R. Head, M. Lee, M. Bernstein, J.A. Squire, A. Smith, P. Dirks, Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. Cell Stem Cell 4, 568–580 (2009)CrossRefGoogle Scholar
- 27.L. Garros-Regulez, P. Aldaz, O. Arrizabalaga, V. Moncho-Amor, E. Carrasco-Garcia, L. Manterola, L. Moreno-Cugnon, C. Barrena, J. Villanua, I. Ruiz, S. Pollard, R. Lovell-Badge, N. Sampron, I. Garcia, A. Matheu, mTOR inhibition decreases SOX2-SOX9 mediated glioma stem cell activity and temozolomide resistance. Expert Opin. Ther. Targets 20, 393–405 (2016)CrossRefPubMedPubMedCentralGoogle Scholar
- 28.J. Popovic, D. Stanisavljevic, M. Schwirtlich, A. Klajn, J. Marjanovic, M. Stevanovic, Expression analysis of SOX14 during retinoic acid induced neural differentiation of embryonal carcinoma cells and assessment of the effect of its ectopic expression on SOXB members in HeLa cells. PLoS One 9, e91852 (2014)CrossRefPubMedPubMedCentralGoogle Scholar
- 30.M. van de Wetering, R. Cavallo, D. Dooijes, M. van Beest, J. van Es, J. Loureiro, A. Ypma, D. Hursh, T. Jones, A. Bejsovec, M. Peifer, M. Mortin, H. Clevers, Armadillo coactivates transcription driven by the product of the Drosophila segment polarity gene dTCF. Cell 88, 789–799 (1997)CrossRefPubMedGoogle Scholar
- 34.T.K. Rimkus, R.L. Carpenter, S. Qasem, M. Chan, H.W. Lo, Targeting the sonic hedgehog signaling pathway: Review of smoothened and GLI inhibitors. Cancers 8, 22 (2016)Google Scholar
- 36.Q.Y. Cai, G.Y. Liang, Y.F. Zheng, Q.Y. Tan, R.W. Wang, K. Li, Sox3 silencing inhibits metastasis and growth of esophageal squamous cell carcinoma cell via down-regulating GSK-3 beta. Int. J. Clin. Exp. Pathol. 9, 2939–2949 (2016)Google Scholar