title: Proteomic analysis of primary cell lines identifies protein changes present in renal cell carcinoma.
abstract: New markers/targets for renal cell carcinoma (RCC) are needed to enable earlier detection and monitoring of disease and therapeutic targeting. To identify such molecules, normal and RCC-derived primary cell lines have been used as a simplified model system for studying changes that accompany tumorigenesis. Short-term cultures allow enrichment of relevant cell types from tissue samples, which is balanced against the potential for in vitro changes. Examination of 3 proteins with altered expression in RCC tissue showed the maintenance of normal-tumour differences in culture, although some changes were apparent, including alteration in the isoform of aldolase. Comparative analysis of primary cell lines by 2-DE found 43 proteins up-regulated and 29 down-regulated in at least three out of five tumour cell lines. Many of the observed changes have been previously reported in RCC, including up-regulation of several glycolytic enzymes, vimentin and heat shock protein 27, validating the approach. Additionally, several novel changes in protein expression were found, including up-regulation of several proteins involved in actin cytoskeleton organisation such as radixin and moesin, two members of the septin family, and the actin bundling protein, fascin. Validation studies using Western blotting and immunohistochemistry indicate that several of these molecules may be useful as markers for RCC.
authors.affiliation: Cancer Research UK Clinical Centre, St. James's University Hospital, Leeds, UK.
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Craven
authors.forename: Anthea J
authors.initials: AJ
authors.lastname: Stanley
authors.forename: Sarah
authors.initials: S
authors.lastname: Hanrahan
authors.forename: James
authors.initials: J
authors.lastname: Dods
authors.forename: Richard
authors.initials: R
authors.lastname: Unwin
authors.forename: Nick
authors.initials: N
authors.lastname: Totty
authors.forename: Patricia
authors.initials: P
authors.lastname: Harnden
authors.forename: Ian
authors.initials: I
authors.lastname: Eardley
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Selby
authors.forename: Rosamonde E
authors.initials: RE
authors.lastname: Banks
mesh_terms: D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000737:chemistry* / Q000378:metabolism*; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000032:analysis / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007672:Kidney Cortex / Q000737:chemistry / Q000378:metabolism; D007680:Kidney Neoplasms / Q000737:chemistry* / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D008840:Microfilament Proteins / Q000032:analysis / Q000378:metabolism; D040901:Proteomics; D000099149:Ezrin
chemical_list: D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D008840:Microfilament Proteins; D000099149:Ezrin; C071058:moesin; C020324:fascin