title: A proteomic approach to analysing spheroid formation of two human thyroid cell lines cultured on a random positioning machine.
abstract: 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.
authors.affiliation: Department of Biology, Chemistry, Pharmacy, FU Berlin, Berlin, Germany.
authors.forename: Jessica
authors.initials: J
authors.lastname: Pietsch
authors.forename: Albert
authors.initials: A
authors.lastname: Sickmann
authors.forename: Gerhard
authors.initials: G
authors.lastname: Weber
authors.forename: Johann
authors.initials: J
authors.lastname: Bauer
authors.forename: Marcel
authors.initials: M
authors.lastname: Egli
authors.forename: Robert
authors.initials: R
authors.lastname: Wildgruber
authors.forename: Manfred
authors.initials: M
authors.lastname: Infanger
authors.forename: Daniela
authors.initials: D
authors.lastname: Grimm
mesh_terms: D000595:Amino Acid Sequence; D000704:Analysis of Variance; D018929:Cell Culture Techniques / Q000295:instrumentation; D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007525:Isoelectric Focusing; D013058:Mass Spectrometry; D008970:Molecular Weight; D020543:Proteome / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism; D013961:Thyroid Gland / Q000378:metabolism*; D013964:Thyroid Neoplasms / Q000378:metabolism*
chemical_list: D003598:Cytoskeletal Proteins; D020543:Proteome