Towards a comprehensive proteome of normal and malignant human colon tissue by 2‐D‐LC‐ESI‐MS and 2‐DE proteomics and identification of S100A12 as potential cancer biomarker

Abstract

The aim of this study was to characterize the proteome of normal and malignant colonic tissue. We previously studied the colon proteome using 2-DE and MALDI-MS and identified 734 proteins (Roeßler, M., Rollinger, W., Palme S., Hagmann, M.-L., et al., Clin. Cancer Res. 2005, 11, 6550-6557).Here we report the identification of additional colon proteins from the same set of tissue samples using a complementary nano-flow 2-D-LC-ESI-MS. In total, 484 proteins were identified in colon. Of these, 252 had also been identified by the 2-DE/MALDI-MS approach, whereas 232 proteins were unique to the 2-D-LC-ESI-MS analysis. Comparing protein expression in neoplastic and normal colon tissue indicated elevated expression of several proteins in colorectal cancer, among them the well established tumor marker carcinoembryonic antigen, as well as calnexin, 40S ribosomal protein S15a, serpin H1, and S100A12. Overexpression of these proteins was confirmed by immunoblotting. Serum levels of S100A12 were determined by ELISA and were found to be strongly elevated in colorectal cancer patients compared to healthy individuals. We conclude, that 2-D-LC-ESI-MS is a powerful approach to identify and compare protein profiles of tissue samples, that it is complementary to 2-DE/MALDI-MS approaches and has the potential to identify novel biomarkers.

Quantitative proteomics analysis of early recurrence/metastasis of huge hepatocellular carcinoma following radical resection

“…Its reported function involves calcium regulation and arachidonic acid metabolism in the neutrophile granulocytes [50]; it has also been reported that the expression of S100A12 was up-regulated both at mRNA and protein levels in the patients with colorectal carcinoma relative to healthy volunteers [62]. The AMACR encoding gene is localized on chromosome 5p13 [51]; it is probably involved in the metabolism of fatty acid and cholalic acid [51].…”

“…The AMACR encoding gene is localized on chromosome 5p13 [51]; it is probably involved in the metabolism of fatty acid and cholalic acid [51]. Previous studies have reported the application of AMACR in the diagnosis of prostatic cancer [62,63] and there are also a few studies that reported AMACR could be used as biomarker to distinguish the hepatocellular carcinoma and the benign tissue [64,65]. However, these two proteins were rarely reported to be associated with the prognosis of HCC.…”

A Combination of Serum Markers for the Early Detection of Colorectal Cancer

“…14), PSME3 (15), and S100A12 (16). The technical performance of the prototype ELISA kits was validated to meet the following specifications (not shown): intraassay CV and interassay CV (<15%), linearity of dilution (AE10% of theoretical value), and analyte recovery in spiked samples (100% AE 20%).…”

Down-regulation of LXR/RXR activation and negative acute phase response pathways in colon adenocarcinoma revealed by proteomics and bioinformatics analysis

“…Previous efforts to identify protein biomarkers in colon cancer tissues or patient fluids have included mass spectrometry, 2D-SDS-PAGE separation and MALDI-TOF identification which are limited by their relatively poor separation and low sensitivity of detection [4,5,25,43]. Though a series of "biomarkers", proteins that are up-regulated or down-regulated in cancerous tissue as compared to healthy tissue, have been established by mass spectrometry, few of them have been established as genuine biomarkers through clinical verification.…”