Nogo‑A/NgR signaling regulates stemness in cancer stem‑like cells derived from U87MG glioblastoma cells

Ai,Chengjin; Zhou,Yu; Pu,Kunming; Yang,Yi; Zhou,Yingying

doi:10.3892/ol.2022.13351

Nogo‑A/NgR signaling regulates stemness in cancer stem‑like cells derived from U87MG glioblastoma cells

Authors:
- Chengjin Ai
- Yu Zhou
- Kunming Pu
- Yi Yang
- Yingying Zhou
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Affiliations: Department of Laboratory Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610072, P.R. China, Department of Ultrasound, The Second People's Hospital of Chengdu, Chengdu, Sichuan 610072, P.R. China, Department of Ultrasound, Chengdu Women's and Children's Central Hospital, School of Medicine, University of Electronic and Technology of China, Chengdu, Sichuan 611731, P.R. China, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 610072, P.R. China
Published online on: May 27, 2022 https://doi.org/10.3892/ol.2022.13351
Article Number: 230
Copyright: © Ai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Neurite outgrowth inhibitor A (Nogo‑A), a member of the reticulon 4 family, is an axon regeneration inhibitor that is negatively associated with the malignancy of oligodendroglial tumors. It has been suggested that the Nogo‑A/Nogo Receptor (NgR) pathway plays a promoting effect in regulating cancer stem‑like cells (CSCs) derived from glioblastoma, indicating that Nogo‑A could exert different roles in CSCs than those in parental cancer cells. In the present study, CSCs were generated from the human Uppsala 87 malignant glioma (U87MG) cell line. These U87MG‑CSCs were characterized by the upregulation of CD44 and CD133, which are two markers of stemness. The expression levels of Nogo‑A and the differentiation of U87MG‑CSCs were investigated. In addition, the proliferation, invasion and colony formation U87MG‑CSCs were examined. Using culture in serum‑containing medium, U87MG‑CSCs were differentiated into neuron‑like cells specifically expressing MAP2, β‑III‑tubulin and nestin. Nogo‑A was upregulated in U87MG‑CSCs compared with parental cells. Knockdown of Nogo‑A and inhibition of the Nogo‑A/NgR signaling pathway in U87MG‑CSCs markedly decreased cell viability, cell cycle entry, invasion and tumor formation, indicating that Nogo‑A could regulate U87MG‑CSC function. Moreover, Nogo‑A was involved in intracellular ATP synthesis and scavenging of accumulated reactive oxygen species. Nogo‑A/NgR pathway exerted protective effects against hypoxia‑induced non‑apoptotic and apoptotic cell death. These results suggest that Nogo‑A plays an important role in regulating U87MG‑CSCs via the Nogo‑A/NgR signaling pathway. Nogo‑A may also different roles in U87MG‑CSCs compared with their parental cells.

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1	Mitre AO, Florian AI, Buruiana A, Boer A, Moldovan I, Soritau O, Florian SI and Susman S: Ferroptosis involvement in glioblastoma treatment. Medicina (Kaunas). 58:3192022. View Article : Google Scholar : PubMed/NCBI
2	Wang Y and Jiang T: Understanding high grade glioma: Molecular mechanism, therapy and comprehensive management. Cancer Lett. 331:139–146. 2013. View Article : Google Scholar
3	Ramaswamy V and Taylor MD: The amazing and deadly glioma race. Cancer Cell. 28:275–277. 2015. View Article : Google Scholar
4	Alcantara Llaguno S and Parada LF: Cancer stem cells in gliomas: Evolving concepts and therapeutic implications. Curr Opin Neurol. 34:868–874. 2021. View Article : Google Scholar
5	Lathia JD, Mack SC, Mulkearns-Hubert EE, Valentim CL and Rich JN: Cancer stem cells in glioblastoma. Genes Dev. 29:1203–1217. 2015. View Article : Google Scholar : PubMed/NCBI
6	Seifert C, Balz E, Herzog S, Korolev A, Gaßmann S, Paland H, Fink MA, Grube M, Marx S, Jedlitschky G, et al: PIM1 inhibition affects glioblastoma stem cell behavior and kills glioblastoma stem-like cells. Int J Mol Sci. 22:111262021. View Article : Google Scholar
7	GrandPré T, Nakamura F, Vartanian T and Strittmatter SM: Identification of the Nogo inhibitor of axon regeneration as a reticulon protein. Nature. 403:439–444. 2000. View Article : Google Scholar
8	Petrinovic MM, Duncan CS, Bourikas D, Weinman O, Montani L, Schroeter A, Maerki D, Sommer L, Stoeckli ET and Schwab ME: Neuronal Nogo-A regulates neurite fasciculation, branching and extension in the developing nervous system. Development. 137:2539–2550. 2010. View Article : Google Scholar
9	Mathis C, Schröter A, Thallmair M and Schwab ME: Nogo-a regulates neural precursor migration in the embryonic mouse cortex. Cereb Cortex. 20:2380–2390. 2010. View Article : Google Scholar : PubMed/NCBI
10	Schwab ME: Functions of Nogo proteins and their receptors in the nervous system. Nat Rev Neurosci. 11:799–811. 2010. View Article : Google Scholar
11	Xiong NX, Zhao HY, Zhang FC and He ZQ: Negative correlation of Nogo-A with the malignancy of oligodendroglial tumor. Neurosci Bull. 23:41–45. 2007. View Article : Google Scholar
12	Schwab DE, Lepski G, Borchers C, Trautmann K, Paulsen F and Schittenhelm J: Immunohistochemical comparative analysis of GFAP, MAP-2, NOGO-A, OLIG-2 and WT-1 expression in WHO 2016 classified neuroepithelial tumors and their prognostic value. Pathol Res Pract. 214:15–24. 2018. View Article : Google Scholar
13	Behling F, Barrantes-Freer A, Behnes CL, Stockhammer F, Rohde V, Adel-Horowski A, Rodríguez-Villagra OA, Barboza MA, Brück W, Lehmann U, et al: Expression of Olig2, nestin, NogoA and AQP4 have no impact on overall survival in IDH-wildtype glioblastoma. PLoS One. 15:e2292742020. View Article : Google Scholar
14	Wang X, Zhou W, Li X, Ren J, Ji G, Du J, Tian W, Liu Q and Hao A: Graphene oxide suppresses the growth and malignancy of glioblastoma stem cell-like spheroids via epigenetic mechanisms. J Transl Med. 18:2002020. View Article : Google Scholar : PubMed/NCBI
15	Shtivelman E and Bishop J: Expression of CD44 is repressed in neuroblastoma cells. Mol Cell Biol. 11:5446–5453. 1991. View Article : Google Scholar
16	Nanduri LS, Maimets M, Pringle SA, van der Zwaag M, van Os RP and Coppes RP: Regeneration of irradiated salivary glands with stem cell marker expressing cells. Radiother Oncol. 99:367–372. 2011. View Article : Google Scholar
17	Wälchli T, Pernet V, Weinmann O, Shiu JY, Guzik-Kornacka A, Decrey G, Yüksel D, Schneider H, Vogel J, Ingber DE, et al: Nogo-A is a negative regulator of CNS angiogenesis. Proc Natl Acad Sci USA. 110:E1943–E1952. 2013. View Article : Google Scholar
18	Wirthschaft P, Bode J, Soni H, Dietrich F, Krüwel T, Fischer B, Knobbe-Thomsen CB, Rossetti G, Hentschel A, Mack N, et al: RhoA regulates translation of the Nogo-A decoy SPARC in white matter-invading glioblastomas. Acta Neuropathol. 138:275–293. 2019. View Article : Google Scholar
19	Aldape K, Zadeh G, Mansouri S, Reifenberger G and von Deimling A: Glioblastoma: Pathology, molecular mechanisms and markers. Acta Neuropathol. 129:829–848. 2015. View Article : Google Scholar
20	Zagrebelsky M and Korte M: Maintaining stable memory engrams: New roles for Nogo-A in the CNS. Neuroscience. 283:17–25. 2014. View Article : Google Scholar : PubMed/NCBI
21	Fang Y, Yao L, Li C, Wang J, Wang J, Chen S, Zhou XF and Liao H: The blockage of the Nogo/NgR signal pathway in microglia alleviates the formation of Aβ plaques and tau phosphorylation in APP/PS1 transgenic mice. J Neuroinflammation. 13:562016. View Article : Google Scholar : PubMed/NCBI
22	Sarkey JP, Chu M, McShane M, Bovo E, Ait Mou Y, Zima AV, de Tombe PP, Kartje GL and Martin JL: Nogo-A knockdown inhibits hypoxia/reoxygenation-induced activation of mitochondrial-dependent apoptosis in cardiomyocytes. J Mol Cell Cardiol. 50:1044–1055. 2011. View Article : Google Scholar
23	Lima FR, Kahn SA, Soletti RC, Biasoli D, Alves T, da Fonseca AC, Garcia C, Romão L, Brito J, Holanda-Afonso R, et al: Glioblastoma: Therapeutic challenges, what lies ahead. Biochim Biophys Acta. 1826:338–349. 2012.
24	Diaz A and Leon K: Therapeutic approaches to target cancer stem cells. Cancers (Basel). 3:3331–3352. 2011. View Article : Google Scholar : PubMed/NCBI
25	Ortensi B, Setti M, Osti D and Pelicci G: Cancer stem cell contribution to glioblastoma invasiveness. Stem Cell Res Ther. 4:182013. View Article : Google Scholar : PubMed/NCBI
26	Safa AR, Saadatzadeh MR, Cohen-Gadol AA, Pollok KE and Bijangi-Vishehsaraei K: Glioblastoma stem cells (GSCs) epigenetic plasticity and interconversion between differentiated non-GSCs and GSCs. Genes Dis. 2:152–163. 2015. View Article : Google Scholar : PubMed/NCBI
27	Meng Y, Shang F and Zhu Y: miR-124 participates in the proliferation and differentiation of brain glioma stem cells through regulating Nogo/NgR expression. Exp Ther Med. 18:2783–2788. 2019.PubMed/NCBI
28	Kern F, Stanika RI, Sarg B, Offterdinger M, Hess D, Obermair GJ, Lindner H, Bandtlow CE, Hengst L and Schweigreiter R: Nogo-A couples with Apg-1 through interaction and co-ordinate expression under hypoxic and oxidative stress. Biochem J. 455:217–227. 2013. View Article : Google Scholar : PubMed/NCBI