title: Differential profiling studies of N-linked glycoproteins in glioblastoma cancer stem cells upon treatment with γ-secretase inhibitor.
abstract: We have recently demonstrated that Notch pathway blockade by γ-secretase inhibitor (GSI) depletes cancer stem cells (CSCs) in Glioblastoma Multiforme (GBM) through reduced proliferation and induced apoptosis. However, the detailed mechanism by which the manipulation of Notch signal induces alterations on post-translational modifications such as glycosylation has not been investigated. Herein, we present a differential profiling work to detect the change of glycosylation pattern upon drug treatment in GBM CSCs. Rapid screening of differential cell surface glycan structures has been performed by lectin microarray on live cells followed by the detection of N-linked glycoproteins from cell lysates using multi-lectin chromatography and label-free quantitative mass spectrometry analysis. A total of 51 and 52 glycoproteins were identified in the CSC- and GSI-treated groups, respectively, filtered by a combination of decoy database searching and Trans-Proteomic Pipeline (TPP) processing. Although no significant changes were detected from the lectin microarray experiment, 7 differentially expressed glycoproteins with high confidence were captured after the multi-lectin column including key enzymes involved in glycan processing. Functional annotations of the altered glycoproteins suggest a phenotype transformation of CSCs toward a less tumorigenic form upon GSI treatment.
authors: Program of Bioinformatics, University of Michigan Medical Center, Ann Arbor, MI 48109-0650, USA.
authors: Lan
authors: L
authors: Dai
authors: Yashu
authors: Y
authors: Liu
authors: Jintang
authors: J
authors: He
authors: Callie G
authors: CG
authors: Flack
authors: Caroline E
authors: CE
authors: Talsma
authors: Jessica G
authors: JG
authors: Crowley
authors: Karin M
authors: KM
authors: Muraszko
authors: Xing
authors: X
authors: Fan
authors: David M
authors: DM
authors: Lubman
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors*; D015153:Blotting, Western; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D020869:Gene Expression Profiling; D005909:Glioblastoma; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000737:chemistry; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D040081:Protein Array Analysis
chemical_list: D004791:Enzyme Inhibitors; D006023:Glycoproteins; D037102:Lectins; D053829:Amyloid Precursor Protein Secretases