title: Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues.
abstract: Renal cell carcinoma (RCC) tissue is composed of a mixture of neoplastic and normal cells, which complicate proteome analysis. The aim of our study was to investigate whether it is feasible to establish primary cell cultures of RCC and of renal cortex maintaining the tissue phenotype along with a more homogeneous and enriched cytological material. Fourteen (82.3%) primary cultures from 17 surgical cases were established and characterized by morphology, growth rate, immunocytochemistry, and molecular analysis performed by Real-time PCR, Western blotting, two-dimensional electrophoresis (2-DE), and mass spectrometry. Cultures showed >90% cytokeratine-positive epithelial cells. In primary tumor cultures, the molecular phenotype of manganese superoxide dismutase and heat shock protein 27 was the same as that found in tumor tissues with overexpression and increased number of isoforms. Moreover, 27 out 28 specific proteins and their isoforms, present in spots excised from 2-DE gel of cortex or RCC cultures, corresponded to those identified on the 2-DE tissue cortex reference map, suggesting that these primary cultures retain the proteomic profile of the corresponding tissues.
authors.affiliation: Department of Experimental, Environmental Medicine and Medical Biotechnologies, Milano-Bicocca University, Monza, Italy. roberto.perego@inimib.it
authors.forename: Roberto A
authors.initials: RA
authors.lastname: Perego
authors.forename: Cristina
authors.initials: C
authors.lastname: Bianchi
authors.forename: Matteo
authors.initials: M
authors.lastname: Corizzato
authors.forename: Barbara
authors.initials: B
authors.lastname: Eroini
authors.forename: Barbara
authors.initials: B
authors.lastname: Torsello
authors.forename: Cristina
authors.initials: C
authors.lastname: Valsecchi
authors.forename: Andrea
authors.initials: A
authors.lastname: Di Fonzo
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Cordani
authors.forename: Paolo
authors.initials: P
authors.lastname: Favini
authors.forename: Stefano
authors.initials: S
authors.lastname: Ferrero
authors.forename: Marina
authors.initials: M
authors.lastname: Pitto
authors.forename: Cecilia
authors.initials: C
authors.lastname: Sarto
authors.forename: Fulvio
authors.initials: F
authors.lastname: Magni
authors.forename: Francesco
authors.initials: F
authors.lastname: Rocco
authors.forename: Paolo
authors.initials: P
authors.lastname: Mocarelli
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000378:metabolism*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000378:metabolism; D007668:Kidney / Q000378:metabolism*; D007680:Kidney Neoplasms / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000378:metabolism; D010449:Peptide Mapping; D010641:Phenotype; D010766:Phosphorylation; D020033:Protein Isoforms; D040901:Proteomics / Q000379:methods*; D012313:RNA / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012694:Serine / Q000737:chemistry; D013482:Superoxide Dismutase / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D018076:DNA, Complementary; D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D020033:Protein Isoforms; D012694:Serine; D012313:RNA; D007633:Keratins; D013482:Superoxide Dismutase