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A proteomic approach to analysing spheroid formation of two human thyroid cell lines cultured on a random positioning machine
Author(s) -
Pietsch Jessica,
Sickmann Albert,
Weber Gerhard,
Bauer Johann,
Egli Marcel,
Wildgruber Robert,
Infanger Manfred,
Grimm Daniela
Publication year - 2011
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000817
Subject(s) - vimentin , spheroid , fibronectin , cytoskeleton , actin , cell culture , cell , myosin , microbiology and biotechnology , proteome , gel electrophoresis , chemistry , biology , two dimensional gel electrophoresis , integrin , intermediate filament , proteomics , biochemistry , genetics , gene , immunology , immunohistochemistry
The human cell lines FTC‐133 and CGTH W‐1, both derived from patients with thyroid cancer, assemble to form different types of spheroids when cultured on a random positioning machine. In order to obtain a possible explanation for their distinguishable aggregation behaviour under equal culturing conditions, we evaluated a proteomic analysis emphasising cytoskeletal and membrane‐associated proteins. For this analysis, we treated the cells by ultrasound, which freed up some of the proteins into the supernatant but left some attached to the cell fragments. Both types of proteins were further separated by free‐flow IEF and SDS gel electrophoresis until their identity was determined by MS. The MS data revealed differences between the two cell lines with regard to various structural proteins such as vimentin, tubulins and actin. Interestingly, integrin α‐5 chains, myosin‐10 and filamin B were only found in FTC‐133 cells, while collagen was only detected in CGTH W‐1 cells. These analyses suggest that FTC‐133 cells express surface proteins that bind fibronectin, strengthening the three‐dimensional cell cohesion.