J Korean Med Sci.  2005 Jun;20(3):450-455. 10.3346/jkms.2005.20.3.450.

Identification of Proteins Differentially Expressed in the Conventional Renal Cell Carcinoma by Proteomic Analysis

• Hwa JS
• Park HJ
• Jung JH
• Kam SC
• Park HC
• Kim CW
• Kang KR
• Hyun JS
• Chung KH

• KMID: 1778505
• DOI: http://doi.org/10.3346/jkms.2005.20.3.450

Abstract

Renal cell carcinoma (RCC) is one of the most malignant tumors in urology, and due to its insidious onset patients frequently have advanced disease at the time of clinical presentation. Thus, early detection is crucial in management of RCC. To identify tumor specific proteins of RCC, we employed proteomic analysis. We prepared proteins from conventional RCC and the corresponding normal kidney tissues from seven patients with conventional RCC. The expression of proteins was determined by silver stain after two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The overall protein expression patterns in the RCC and the normal kidney tissues were quite similar except some areas. Of 66 differentially expressed protein spots (p<0.05 by Student t-test), 8 different proteins from 11 spots were identified by MALDI-TOF-MS. The expression of the following proteins was repressed (p<0.05); aminoacylase-1, enoyl-CoA hydratase, aldehyde reductase, tropomyosin alpha-4 chain, agmatinase and ketohexokinase. Two proteins, vimentin and alpha-1 antitrypsin precursor, were dominantly expressed in RCC (p<0.05).

Keyword

Kidney; Carcinoma, Renal Cell; Electrophoresis, Gel, Two-Dimensional; Proteomics; Vimentin; Alpha 1-Antitrypsin

MeSH Terms

Aged
Aldehyde Reductase/analysis
Amidohydrolases/analysis
Carcinoma, Renal Cell/*metabolism/pathology
Comparative Study
Electrophoresis, Gel, Two-Dimensional
Enoyl-CoA Hydratase/analysis
Female
Fructokinases/analysis
Humans
Kidney Neoplasms/*metabolism/pathology
Male
Middle Aged
Proteome/*analysis
Proteomics/*methods
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Tropomyosin/analysis
Ureohydrolases/analysis
Vimentin/analysis
alpha 1-Antitrypsin/analysis

Figure

• 

  Fig. 1 Two-dimensional gel analysis of proteins from the conventional RCC tissue (A) and the corresponding normal kidney tissue (B). Isoelectric focusing at pH 4-7 was carried out at the first dimension using IPG strips, and a 7.5-17.5% gradient SDS gel electrophoresis was followed at the second dimension as described in "Materials and methods". Numbers represent region of interest that were identified by mass spectrometry and peptide mass fingerprinting.

• 

  Fig. 2 Typical spectrum of Aldehyde reductase obtained from peptide mass fingerprinting of a gel spot. After separation of the proteins by two-dimensional gel electrophoresis, the gel spot was excised and digested with trypsin. Peptides obtained from digestion were analyzed by MALDI-TOF/mass spectrometry. The abscissa represents the mass/charge ratio of ions detected in the MALDI-TOF experiments. Peaks are almost always protonated molecular ion [M+H+]. The ordinate represents the relative intensity of the averaged ion current in arbitrary units.

• 

  Fig. 3 The intensity of protein spots (spot intensity from PDQuest software) down-regulated in the conventional RCC samples. Differentially expressed protein spots identified by peptide mass fingerprinting are labeled with protein name. The graph shows difference of spot density as compared normal kidney tissues (closed bar) with RCC (shaded bar). AVE, average.

• 

  Fig. 4 The intensity of protein spots up-regulated in the conventional RCC samples. Differentially expressed protein spots identified by peptide mass fingerprinting are labeled with protein name. The graph shows difference of spot density as compared normal kidney tissues (closed bar) with RCC (shaded bar).

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