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Colon cancer - Characterization of changes in the proteome in different regions of 3D multicell tumor spheroids

ABSTRACT: Proteins were extracted from different regions of HT29 colon carcinoma cells grown as multicell tumour spheroids. Changes in protein expression across the regions were determined by trypsin digestion iTRAQ 4-plex labelling, 2D separation using OffGel (24 fractions) and RP nanoHPLC, MALDI TOF-TOF MS/MS analysis and Mascot database searching.

INSTRUMENT(S): Bruker Daltonics flex series, instrument model

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Colonic Cancer Cell Line, Colon

SUBMITTER: Chris Sutton

PROVIDER: PRD000369 | Pride | 2014-04-08

REPOSITORIES: Pride

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Characterization of changes in the proteome in different regions of 3D multicell tumor spheroids.

McMahon K M KM Volpato M M Chi H Y HY Musiwaro P P Poterlowicz K K Peng Y Y Scally A J AJ Patterson L H LH Phillips R M RM Sutton C W CW

Journal of proteome research 20120330 5

Three dimensional multicell tumor spheroids (MCTS) provide an experimental model where the influence of microenvironmental conditions on protein expression can be determined. Sequential trypsin digestion of HT29 colon carcinoma MCTS enabled segregation into four populations comprising proliferating cells from the surface (SL), an intermediate region (IR), nonproliferating hypoxic cells from the perinecrotic region (PN), and a necrotic core (NC). Total protein was extracted from each population a ...[more]

PMID: 22416669

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