Dataset Information

An insight into the proteome of uveal melanoma-derived ectosomes reveals the presence of potentially useful biomarkers

ABSTRACT: Cancer cells are known to release extracellular vesicles that often promote disease development and progression. The present study investigated protein content and glycosylation pattern of ectosomes released in vitro by human primary uveal melanoma Mel202 cell line. Ectosomes released by Mel202 cells were isolated from conditioned media by sequential centrifugation and nanoLC-MS/MS approach was used to determine their protein content. Subsequently, proteins from ectosomes, the whole cell extracts and the membrane fractions were probed with a panel of lectins to reveal characteristic glycan structures. As many as 2529 unique proteins were identified, and many of them are known to be involved in cancer cell proliferation and altered metabolism, tumor invasion, metastasis or drug resistance. Lectin-based studies showed a distinct glycosylation pattern between Mel202 ectosomes and parental cell membranes. Selective enrichment of ectosomal proteins with bisected complex type N-glycans and α2,6-linked sialic acid may be significant for ectosome formation and sequestration. Our data supports recent findings that ectosomes derive from particular regions of the cell membrane and thus contain a unique protein and glycan composition. Exploring whether cancer-related proteins and glycans are enriched in ectosomes isolated from body fluids of cancer patients should became a subject of future in-depth studies.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Urszula Jankowska

LAB HEAD: Sylwia Kedracka-Krok

PROVIDER: PXD012003 | Pride | 2019-08-08

REPOSITORIES: pride

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Dataset's files

Source:

			Action	DRS
	17-07-29_Hoja_Ektosomy_1-01.msf	Msf
	17-07-29_Hoja_Ektosomy_1.raw	Raw
	18_07_03_ektosomy_Przybylo.msf	Msf
	18_07_03_ektosomy_Przybylo.raw	Raw
	18_07_04_ektosomy_Przybylo_1.msf	Msf

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Publications

An Insight into the Proteome of Uveal Melanoma-Derived Ectosomes Reveals the Presence of Potentially Useful Biomarkers.

Surman Magdalena M Hoja-Łukowicz Dorota D Szwed Sabina S Kędracka-Krok Sylwia S Jankowska Urszula U Kurtyka Magdalena M Drożdż Anna A Lityńska Anna A Stępień Ewa E Przybyło Małgorzata M

International journal of molecular sciences 20190802 15

Cancer cells are known to release extracellular vesicles that often promote disease development and progression. The present study investigated the protein content and glycosylation pattern of ectosomes released in vitro by a human primary uveal melanoma Mel202 cell line. Ectosomes released by Mel202 cells were isolated from conditioned media using sequential centrifugation, and a nano-LC-MS/MS approach was used to determine their protein content. Subsequently, proteins from ectosomes, the whole ...[more]

PMID: 31382537

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Project description:This data set was downloaded from MetaboLights (http://www.ebi.ac.uk/metabolights/) accession number MTBLS227 Abstract:"Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. However, differences between CRC cell lines with regard to their glycosylation have hitherto been largely neglected. Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. Released glycans were derivatized to stabilize and differentiate between a2,3- and a2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation."