title: Differential patterns of NOTCH1-4 receptor expression are markers of glioma cell differentiation.
abstract: Background Notch signaling is deregulated in human gliomas and may play a role in their malignancy. However, the role of each Notch receptor in glioma cell differentiation and progression is not clear. We examined the expression pattern of Notch receptors and compared it with differentiation markers in glioma cell lines, primary human cultures, and biopsies of different grades. Furthermore, the effects of a γ-secretase inhibitor (GSI) on cell survival were assessed. Methods Notch receptors and markers of cellular differentiation were analyzed by reverse transcriptase PCR, Western blotting, immunohistochemistry, and immunocytochemistry. GSI sensitivity was assessed in both cell lines and primary cultures grown as monolayers or tumorspheres, by MTT assay. Results In cell lines, Notch1 and Notch2/4 levels paralleled those of glial fibrillary acidic protein (GFAP) and vimentin, respectively. In human gliomas and primary cultures, Notch1 was moderate/strong in low-grade tumors but weak in glioblastoma multiforme (GBM). Conversely, Notch4 increased from astrocytoma grade II to GBM. Primary GBM cultures grown in serum (monolayer) showed moderate/high levels of CD133, nestin, vimentin, and Notch4 and very low levels of GFAP and Notch1, which were reduced in tumorspheres. This effect was drastic for Notch4. GSI reduced cell survival with stronger effect in serum, whilst human primary cultures showed different sensitivity. Conclusion Data from cell lines and human gliomas suggest a correlation between expression of Notch receptors and cell differentiation. Namely, Notch1 and Notch4 are markers of differentiated and less differentiated glioma cells, respectively. We propose Notch receptors as markers of glioma grading and possible prognostic factors.
authors.affiliation: Corresponding authors: Paola Dell'Albani, PhD, Institute of Neurological Sciences, CNR, Via P. Gaifami, 18-95126 Catania, Italy. paola.dellalbani@cnr.it); Maria Vincenza Catania, MD, Institute of Neurological Sciences, CNR, Via P. Gaifami, 18-95126 Catania, Italy (mariavincenza.catania@cnr.it.
authors.forename: Paola
authors.initials: P
authors.lastname: Dell'albani
authors.forename: Margherita
authors.initials: M
authors.lastname: Rodolico
authors.forename: Rosalia
authors.initials: R
authors.lastname: Pellitteri
authors.forename: Elisa
authors.initials: E
authors.lastname: Tricarichi
authors.forename: Sebastiano Alfio
authors.initials: SA
authors.lastname: Torrisi
authors.forename: Simona
authors.initials: S
authors.lastname: D'Antoni
authors.forename: Mario
authors.initials: M
authors.lastname: Zappia
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Albanese
authors.forename: Rosario
authors.initials: R
authors.lastname: Caltabiano
authors.forename: Nunzio
authors.initials: N
authors.lastname: Platania
authors.forename: Eleonora
authors.initials: E
authors.lastname: Aronica
authors.forename: Maria Vincenza
authors.initials: MV
authors.lastname: Catania
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors; D000818:Animals; D017209:Apoptosis; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005910:Glioma / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007124:Immunoenzyme Techniques; D060787:Neoplasm Grading; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000235:genetics / Q000378:metabolism; D051883:Receptor, Notch2 / Q000235:genetics / Q000378:metabolism; D000071656:Receptor, Notch3; D000074562:Receptor, Notch4; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D014408:Biomarkers, Tumor; D004791:Enzyme Inhibitors; C499374:NOTCH3 protein, human; C106075:NOTCH4 protein, human; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D051881:Receptor, Notch1; D051883:Receptor, Notch2; D000071656:Receptor, Notch3; D000074562:Receptor, Notch4; D051880:Receptors, Notch; D053829:Amyloid Precursor Protein Secretases
keywords: Notch receptors; cell differentiation; gene expression; glioma cell cultures; gliomas; intermediate filaments