Abstract
BackgroundCancer stem cells (CSCs) are reported to be responsible for tumor initiation, progression, metastasis, and therapy resistance where P-glycoprotein (P-gp) as well as other glycoproteins are involved. Identification of these glycoprotein markers is critical for understanding the resistance mechanism and developing therapeutics.MethodsIn this study, we report our comparative and quantitative site- and structure-specific N-glycoproteomics study of MCF-7/ADR cancer stem cells (CSCs) vs. MCF-7/ADR cells. With zic-HILIC enrichment, isotopic diethyl labeling, RPLC–MS/MS (HCD) analysis and GPSeeker DB search, differentially expressed N-glycosylation was quantitatively characterized at the intact N-glycopeptide level.Results4016 intact N-glycopeptides were identified with spectrum-level FDR ≤ 1%. With the criteria of ≥ 1.5 fold change and p value < 0.05, 247 intact N-glycopeptides were found differentially expressed in MCF-7/ADR CSCs as putative markers. Raw data are available via ProteomeXchange with identifier PXD013836.ConclusionsQuantitative site- and structure-specific N-glycoproteomics characterization may help illustrate the cell stemness property.
Highlights
Aberrant N-glycosylation is increasingly recognized as one of the most important biochemical changes involved in tumorigenesis and metastasis [1,2,3,4]; most of the FDA approved cancer biomarkers are glycoproteins
In 2017, Xing et al found that ALDH1 and ABCG2 were enhanced in primary foci and metastatic lymph node from patients with triple-negative breast cancer with qRT-PCR, western blotting and MTT assay of mRNA expression, protein expression and proliferation of MDAMB-231 cells, respectively; and the authors proposed that ALDH1 and ABCG2 may affect the drug resistance [13]
In 2018, Bogusha et al found that induction of the epithelial-mesenchymal transition process made bigger contribution to the drug resistance of MCF-7/ADR cells than the ATP-binding cassette (ABC) transporter’s overexpression, because differential expression of vimentin is much higher than that of P-gp as measured by immunofluorescent staining with antibodies [14]
Summary
We report our comparative and quantitative site- and structure-specific N-glycoproteomics study of MCF-7/ADR cancer stem cells (CSCs) vs. MCF-7/ADR cells. With zic-HILIC enrichment, isotopic diethyl labeling, RPLC–MS/MS (HCD) analysis and GPSeeker DB search, differentially expressed N-glycosylation was quantitatively characterized at the intact N-glycopeptide level
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