Structural N-Glycoproteomics Characterization of Cell-Surface N-Glycosylation of MCF-7/ADR Cancer Stem Cells

Abstract

Breast cancer is responsible for the highest mortality all over the world. Cancer stem cells (CSCs) along with epithelial mesenchymal transition (EMT) are identified as a driver of cancer which are responsible for cancer metastasis and drug resistance. Several signaling pathways are associated with drug resistance. Additionally, glycosyltransferases regulate different types of glycosylation which are involved in drug resistance. To the end, it is urgent to figure out the knowledge on cell-surface altered N-glycosylation and putative markers. Here, differential cell-surface intact N-glycopeptides in adriamycin (ADR)-resistant michigan breast cancer foundation-7 stem cells (MCF-7/ADR CSCs) relative to ADR-sensitive MCF-7 CSCs were analyzed with site- and structure-specific quantitative N-glycoproteomics. The intact N-glycopeptides and differentially expressed intact N-glycopeptides (DEGPs) were determined and quantified via intact N-glycopeptide search engine GPSeeker. Totally, 4,777 intact N-glycopeptides were identified and N-glycan sequence structures among 2,764 IDs were distinguished from their isomers by structure-diagnostic fragment ions. Among 1,717 quantified intact N-glycopeptides, 104 DEGPs were determined (fold change≥1.5 and p value<0.05). Annotation of protein-protein interaction and biological processes among others of DEGPs were finally carried out; down-regulated intact N-glycopeptide with bisecting GlcNAc from p38-interacting protein and up-regulated intact N-glycopeptide with β1,6-branching N-glycan from integrin beta-5 were found.

Note:
Funding declaration: This research was supported by the National Science Foundation of China (22074105) and the Shanghai Science and Technology Commission (14DZ2261100).

Conflict of Interests: The authors declare no competing interests.

Keywords: cell-surface, drug resistance, cancer stem cell, MCF-7, quantitative N-glycoproteomics