Glycoproteomic analysis of glioblastoma stem cell differentiation.
Cancer stem cells are responsible for tumor formation through self-renewal and differentiation into multiple cell types and thus represent a new therapeutic target for tumors. Glycoproteins play a critical role in determining the fates of stem cells such as self-renewal, proliferation, and differentiation. Here we applied a multilectin affinity chromatography and quantitative glycoproteomics approach to analyze alterations of glycoproteins relevant to the differentiation of a glioblastoma-derived stem cell line HSR-GBM1. Three lectins including concanavalin A (Con A), wheat germ agglutinin (WGA), and peanut agglutinin (PNA) were used to capture glycoproteins, followed by LC-MS/MS analysis. A total of 73 and 79 high-confidence (FDR < 0.01) glycoproteins were identified from the undifferentiated and differentiated cells, respectively. Label-free quantitation resulted in the discovery of 18 differentially expressed glycoproteins, wherein 9 proteins are localized in the lysosome. All of these lysosomal glycoproteins were up-regulated after differentiation, where their principal function was hydrolysis of glycosyl residues. Protein-protein interaction and functional analyses revealed the active involvement of lysosomes during the process of glioblastoma stem cell differentiation. This work provides glycoprotein markers to characterize differentiation status of glioblastoma stem cells that may be useful in stem-cell therapy of glioblastoma.
Department of Surgery, University of Michigan Medical Center, Ann Arbor, Michigan 48109, USA.
Jintang
J
He
Yashu
Y
Liu
Thant S
TS
Zhu
Xiaolei
X
Xie
Mark A
MA
Costello
Caroline E
CE
Talsma
Callie G
CG
Flack
Jessica G
JG
Crowley
Francesco
F
Dimeco
Angelo L
AL
Vescovi
Xing
X
Fan
David M
DM
Lubman
D015153:Blotting, Western; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D048430:Cell Shape; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D016000:Cluster Analysis; D030562:Databases, Protein; D005909:Glioblastoma / Q000737:chemistry* / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D014411:Neoplastic Stem Cells; D037121:Plant Lectins / Q000378:metabolism; D025941:Protein Interaction Mapping; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry
D006023:Glycoproteins; D037121:Plant Lectins