Identification of cell surface glycoprotein markers for glioblastoma-derived stem-like cells using a lectin microarray and LC-MS/MS approach.
Despite progress in the treatment of glioblastoma, more than 95% of patients suffering from this disease still die within 2 years. Recent findings support the belief that cancer stem-like cells are responsible for tumor formation and ongoing growth. Here a method combining lectin microarray and LC-MS/MS was used to discover the cell surface glycoprotein markers of a glioblastoma-derived stem-like cell line. Lectin microarray analysis of cell surface glycans showed that two galactose-specific lectins Trichosanthes kirilowii agglutinin (TKA) and Peanut agglutinin (PNA) could distinguish the stem-like glioblastoma neurosphere culture from a traditional adherent glioblastoma cell line. Agarose-bound TKA and PNA were used to capture the glycoproteins from the two cell cultures, which were analyzed by LC-MS/MS. The glycoproteins were quantified by spectral counting, resulting in the identification of 12 and 11 potential glycoprotein markers from the TKA and PNA captured fractions respectively. Almost all of these proteins were membrane proteins. Differential expression was verified by Western blotting analysis of 6 interesting proteins, including the up-regulated Receptor-type tyrosine-protein phosphatase zeta, Tenascin-C, Chondroitin sulfate proteoglycan NG2, Podocalyxin-like protein 1 and CD90, and the down-regulated CD44. An improved understanding of these proteins may be important for earlier diagnosis and better therapeutic targeting of glioblastoma.
Department of Surgery, University of Michigan Medical Center, Ann Arbor, Michigan 48109, USA.
Jintang
J
He
Yashu
Y
Liu
Xiaolei
X
Xie
Thant
T
Zhu
Mary
M
Soules
Francesco
F
DiMeco
Angelo L
AL
Vescovi
Xing
X
Fan
David M
DM
Lubman
D000595:Amino Acid Sequence; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D005909:Glioblastoma / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000378:metabolism; D008969:Molecular Sequence Data; D014411:Neoplastic Stem Cells / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry* / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D013347:Subcellular Fractions / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods
D014408:Biomarkers, Tumor; D008562:Membrane Glycoproteins; D037121:Plant Lectins