Glioblastoma is the most common malignant brain tumor accounting for more than 54 % of all gliomas. Despite aggressive treatments, median survival remains less than 1 year. This might be due to the unavailability of effective molecular diagnostic markers and targeted therapy. Thus, it is essential to discover molecular mechanisms underlying disease by identifying dysregulated pathways involved in tumorigenesis. Notch signaling is one such pathway which plays an important role in determining cell fates. Since it is found to play a critical role in many cancers, we investigated the role of Notch genes in glioblastoma with an aim to identify biomarkers that can improve diagnosis. Using real-time PCR, we assessed the expression of Notch genes including receptors (Notch1, Notch2, Notch3, and Notch4), ligands (JAG1, JAG2, and DLL3), downstream targets (HES1 and HEY2), regulator Deltex1 (DTX1), inhibitor NUMB along with transcriptional co-activator MAML1, and a component of gamma-secretase complex APH1A in 15 formalin-fixed paraffin-embedded (FFPE) patient samples. Relative quantification was done by the 2−ΔΔCt method; the data are presented as fold change in gene expression normalized to an internal control gene and relative to the calibrator. The data revealed aberrant expression of Notch genes in glioblastoma compared to normal brain. More than 85 % of samples showed high Notch1 (P = 0.0397) gene expression and low HES1 (P = 0.011) and DTX1 (P = 0.0001) gene expression. Our results clearly show aberrant expression of Notch genes in glioblastoma which can be used as putative biomarkers together with histopathological observation to improve diagnosis, therapeutic strategies, and patient prognosis.
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This study was funded by the Department of Science and Technology, New Delhi, India (grant number SR/FT/LS-170/2008). We thank the Brain Bank, NIMHANS, Bangalore, India, for providing the clinical samples.
Compliance with ethical standards
Conflicts of interest
For this type of study, formal consent is not required.
Ostrom QT, Gittleman H, Liao P, et al. CBTRUS statistical report: primary brain and central nervous system tumors diagnosed in the United States in 2007-2011. Neuro Oncol. 2014;16 Suppl 4:iv1–63.CrossRefPubMedPubMedCentralGoogle Scholar
Mischel PS, Vinters HV. Neuropathology and molecular pathogenesis of primary brain tumors. Brain Tumor Immunother. 2001;3–45.Google Scholar
Godard S, Getz G, Delorenzi M, et al. Classification of human astrocytic gliomas on the basis of gene expression: a correlated group of genes with angiogenic activity emerges as a strong predictor of subtypes. Cancer Res. 2003;63:6613–25.PubMedGoogle Scholar
Nutt CL, Mani DR, Betensky RA, et al. Gene expression-based classification of malignant gliomas correlates better with survival than histological classification. Cancer Res. 2003;63:1602–7.PubMedGoogle Scholar
Furukawa T, Mukherjee S, Bao ZZ, et al. rax, Hes1, and Notch1 promote the formation of Müller glia by postnatal retinal progenitor cells. Neuron. 2000;26:383–94.CrossRefPubMedGoogle Scholar
Gaiano N, Fishell G. The role of Notch in promoting glial and neural stem cell fates. Annu Rev Neurosci. 2002;25:471–90.CrossRefPubMedGoogle Scholar
Li HS, Wang D, Shen Q, et al. Inactivation of Numb and Numblike in embryonic dorsal forebrain impairs neurogenesis and disrupts cortical morphogenesis. Neuron. 2003;40:1105–18.CrossRefPubMedGoogle Scholar
Baumgart A, Mazur PK, Anton M, et al. Opposing role of Notch1 and Notch2 in a KrasG12D-driven murine non-small cell lung cancer model. Oncogene. 2015;34:578–88.CrossRefPubMedGoogle Scholar
Purow BW, Haque RM, Noel MW, et al. Expression of Notch-1 and its ligands, Delta-like-1 and Jagged-1, is critical for glioma cell survival and proliferation. Cancer Res. 2005;65:2353–63.CrossRefPubMedGoogle Scholar
Zhang XP, Zheng G, Zou L, et al. Notch activation promotes cell proliferation and the formation of neural stem cell-like colonies in human glioma cells. Mol Cell Biochem. 2008;307:101–8.CrossRefPubMedGoogle Scholar
Hovinga KE, Shimizu F, Wang R, et al. Inhibition of notch signaling in glioblastoma targets cancer stem cells via an endothelial cell intermediate. Stem Cells. 2010;28:1019–29.CrossRefPubMedGoogle Scholar
Ridgway J, Zhang G, Wu Y, et al. Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis. Nature. 2006;444:1083–7.CrossRefPubMedGoogle Scholar
Kizys MM, Cardoso MG, Lindsey SC, et al. Optimizing nucleic acid extraction from thyroid fine-needle aspiration cells in stained slides, formalin-fixed/paraffin-embedded tissues, and long-term stored blood samples. Arq Bras Endocrinol Metabol. 2012;56:618–26.CrossRefPubMedGoogle Scholar
Aithal MGS, Rajeswari N. Validation of housekeeping genes for gene expression analysis in glioblastoma using quantitative real-time polymerase chain reaction. Brain Tumor Res Treat. 2015;3:24–9.CrossRefPubMedPubMedCentralGoogle Scholar
Sasnauskienė A, Jonušienė V, Krikštaponienė A, et al. NOTCH1, NOTCH3, NOTCH4, and JAG2 protein levels in human endometrial cancer. Medicina (Kaunas). 2014;50:14–8.CrossRefGoogle Scholar
Ye QF, Zhang YC, Peng XQ, et al. Silencing Notch-1 induces apoptosis and increases the chemosensitivity of prostate cancer cells to docetaxel through Bcl-2 and Bax. Oncol Lett. 2012;3:879–84.PubMedPubMedCentralGoogle Scholar
Wei G, Chang Y, Zheng J, et al. Notch1 silencing inhibits proliferation and invasion in SGC-7901 gastric cancer cells. Mol Med Rep. 2014;9:1153–8.PubMedGoogle Scholar
Chen AC, Guo LY, Ostaszewski BL, et al. Aph-1 associates directly with full-length and C-terminal fragments of gamma-secretase substrates. J Biol Chem. 2010;285:11378–91.CrossRefPubMedPubMedCentralGoogle Scholar
Bae SK, Bessho Y, Hojo M, et al. The bHLH gene Hes6, an inhibitor of Hes1, promotes neuronal differentiation. Development. 2000;127:2933–43.PubMedGoogle Scholar
Gibert JM, Simpson P. Evolution of cis-regulation of the proneural genes. Int J Dev Biol. 2003;47:643–51.PubMedGoogle Scholar
Luo B, Aster JC, Hasserjian RP, et al. Isolation and functional analysis of a cDNA for human Jagged2, a gene encoding a ligand for the Notch1 receptor. Mol Cell Biol. 1997;17:6057–67.CrossRefPubMedPubMedCentralGoogle Scholar
Wu L, Griffin JD. Modulation of Notch signaling by mastermind-like (MAML) transcriptional co-activators and their involvement in tumorigenesis. Semin Cancer Biol. 2004;14:348–56.CrossRefPubMedGoogle Scholar
Zhang P, Yang Y, Nolo R, et al. Regulation of NOTCH signaling by reciprocal inhibition of HES1 and Deltex 1 and its role in osteosarcoma invasiveness. Oncogene. 2010;29:2916–26.CrossRefPubMedPubMedCentralGoogle Scholar
Izon DJ, Aster JC, He Y, et al. Deltex1 redirects lymphoid progenitors to the B cell lineage by antagonizing Notch1. Immunity. 2002;16:231–43.CrossRefPubMedGoogle Scholar