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Primary cell cultures arising from normal kidney and renal cell carcinoma retain the proteomic profile of corresponding tissues.
- Source :
-
Journal of proteome research [J Proteome Res] 2005 Sep-Oct; Vol. 4 (5), pp. 1503-10. - Publication Year :
- 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.
- Subjects :
- Adult
Aged
Aged, 80 and over
Blotting, Western
Cell Line, Tumor
Cells, Cultured
DNA, Complementary metabolism
Electrophoresis, Agar Gel
Electrophoresis, Gel, Two-Dimensional
Epithelial Cells metabolism
Female
HSP27 Heat-Shock Proteins
Heat-Shock Proteins metabolism
Humans
Immunohistochemistry
Keratins metabolism
Male
Mass Spectrometry
Middle Aged
Molecular Chaperones
Neoplasm Proteins metabolism
Peptide Mapping
Phenotype
Phosphorylation
Protein Isoforms
RNA chemistry
Reverse Transcriptase Polymerase Chain Reaction
Serine chemistry
Superoxide Dismutase metabolism
Tumor Cells, Cultured
Carcinoma, Renal Cell metabolism
Gene Expression Regulation, Neoplastic
Kidney metabolism
Kidney Neoplasms metabolism
Proteomics methods
Subjects
Details
- Language :
- English
- ISSN :
- 1535-3893
- Volume :
- 4
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of proteome research
- Publication Type :
- Academic Journal
- Accession number :
- 16212400
- Full Text :
- https://doi.org/10.1021/pr050002o