Modified expression of plasma glutathione peroxidase and manganese superoxide dismutase in human renal cell carcinoma

Abstract

Two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) is a powerful tool to separate thousands of polypeptides and to highlight the modification of protein expression in malignant diseases. By applying 2‐D PAGE to ten normal human kidney and ten homologous renal cell carcinoma (RCC) tissues, we found two peptides in all ten normal tissues but not in RCCs and, conversely, two peptides were detected in all RCCs but not in normal tissues. Using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) and internal sequence analysis, the two first peptides were identified as two isoforms of plasma glutathione peroxidase (GPxP). The two other peptides isolated in all RCCs but not in normal tissues were identified by N‐terminal sequence analysis as multimeric forms of manganese superoxide dismutase (Mn‐SOD). No multimeric Mn‐SODs and only two monomeric forms were detected in normal tissues. GPxP and Mn‐SOD are metallo‐enzymes encoded on chromosome 5q32 and on chromosome 6p25, respectively. Their regions are within the locus 5q21 → qter and 6q21‐6q27 on which deletions and translocations are described in some cytogenetic studies of RCC transformation. Therefore, our results might suggest a correlation between the modified expression of GPxP and Mn‐SOD in tumor tissues and chromosomal modifications, and that the two proteins may be putative markers for diagnosis of RCC.*

Contribution of proteomics to the molecular analysis of renal cell carcinoma with an emphasis on manganese superoxide dismutase

“…Heterogeneity of RCC cell lines was also revealed using 2-DE, highlighting different levels in the expression of proliferating cell nuclear antigen and glutathione-Stransferase [50]. Specific antibodies to epithelial antigens bound to magnetic beads can be used to improve separation of epithelial cells [55][56], but laser capture microdissection is a more promising technique to resolve the problem of contamination and mixture of cell types [57]. In addition, to improve the RCC protein characterization by 2-D PAGE, Starita-Geribaldi et al [58] described a protocol to analyse specific subcellular fractions such as basolateral membranes containing protease SP220K, a potential RCC marker.…”

Targeting of tumor associated antigens in renal cell carcinoma using proteome-based analysis and their clinical significance

“…Recent reports suggest that 2-DE based proteome analysis or PROTEOMEX, a combination of conventional proteome analysis with serology, might allow the identification of diagnostic and prognostic markers in RCC [5][6][7][8][9][10]. 2-DE patterns of normal renal parenchyma and RCC samples revealed a number of polypeptides functional associated with RCC [5,[11][12][13]. These include proteins of mitochondrial pathway, such as the ubiquinol cytochrome reductase and the mitochondrial NADH-ubiquinone oxidoreductase complex I, which are expressed in normal kidney tissues, but absent in RCC lesions [11] and enzymes that play an important role in controlling redox changes and detoxification and enzymes of the carbohydrate metabolism [12,13].…”

“…2-DE patterns of normal renal parenchyma and RCC samples revealed a number of polypeptides functional associated with RCC [5,[11][12][13]. These include proteins of mitochondrial pathway, such as the ubiquinol cytochrome reductase and the mitochondrial NADH-ubiquinone oxidoreductase complex I, which are expressed in normal kidney tissues, but absent in RCC lesions [11] and enzymes that play an important role in controlling redox changes and detoxification and enzymes of the carbohydrate metabolism [12,13]. In addition, the serological based proteome analysis also resulted in the definition of tumor-associated markers in RCC [6,14].…”

Mapping and expression pattern analysis of key components of the major histocompatibility complex class I antigen processing and presentation pathway in a representative human renal cell carcinoma cell line

“…Several strategies exist to define potential target structures for this therapeutic modality, including 2-D PAGE separation [30,33], serological identification of antigens by recombinant expression cloning (SEREX) analysis [34], cDNA expression cloning [35], and subtractive hybridization procedures [36]. The frequent occurrence of immune escape variants within tumor entities has rekindled interest for the mapping of key components of the MHC class I antigen processing and presentation pathway, which might be helpful in the evaluation and selection of patients to be treated with CTL-based immunotherapeutic approaches.…”