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.

Perego, R., Bianchi, C., Corizzato, M., Eroini, B., Torsello, B., Valsecchi, C., et al. (2005). Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues. JOURNAL OF PROTEOME RESEARCH, 4(5), 1503-1510 [10.1021/pr050002o].

Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues

PEREGO, ROBERTO;BIANCHI, CRISTINA;CORIZZATO, MATTEO;EROINI, BARBARA;TORSELLO, BARBARA ROSA;DI FONZO, ANDREA;CORDANI, NICOLETTA;PITTO, MARINA;MAGNI, FULVIO;MOCARELLI, PAOLO
2005

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.
Articolo in rivista - Articolo scientifico
Heat-Shock Proteins; Epithelial Cells; Electrophoresis, Agar Gel; Gene Expression Regulation, Neoplastic; Middle Aged; Electrophoresis, Gel, Two-Dimensional; Neoplasm Proteins; Immunohistochemistry; Blotting, Western; Cell Line, Tumor; HSP27 Heat-Shock Proteins; Tumor Cells, Cultured; Phosphorylation; Aged; Mass Spectrometry; Adult; DNA, Complementary; Kidney; Aged, 80 and over; Male; Cells, Cultured; RNA; Proteomics; Keratins; Female; Kidney Neoplasms; Phenotype; Serine; Humans; Peptide Mapping; Protein Isoforms; Reverse Transcriptase Polymerase Chain Reaction; Carcinoma, Renal Cell; Superoxide Dismutase
English
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Perego, R., Bianchi, C., Corizzato, M., Eroini, B., Torsello, B., Valsecchi, C., et al. (2005). Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues. JOURNAL OF PROTEOME RESEARCH, 4(5), 1503-1510 [10.1021/pr050002o].
Perego, R; Bianchi, C; Corizzato, M; Eroini, B; Torsello, B; Valsecchi, C; DI FONZO, A; Cordani, N; Favini, P; Ferrero, S; Pitto, M; Sarto, C; Magni, F; Rocco, F; Mocarelli, P
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/12615
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