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

Roberto A. Perego, Cristina Bianchi, Matteo Corizzato, Barbara Eroini, Barbara Torsello, Cristina Valsecchi, Andrea Di Fonzo, Nicoletta Cordani, Paolo Favini, Stefano Ferrero, Marina Pitto, Cecilia Sarto, Fulvio Magni, Francesco Rocco, Paolo Mocarelli

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)1503-1510
Number of pages8
JournalJournal of Proteome Research
Volume4
Issue number5
DOIs
Publication statusPublished - Sep 2005

Keywords

  • Heat shock protein 27
  • Immunocytochemistry
  • Manganese superoxide dismutase
  • Mass spectrometry
  • Primary cultures
  • Proteome
  • Real-time PCR
  • Renal cell carcinoma
  • Two-dimensional gel electrophoresis
  • Western blotting

ASJC Scopus subject areas

  • Genetics
  • Biotechnology
  • Biochemistry

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