Identification of PSME3 as a novel serum tumor marker for colorectal cancer by combining two-dimensional polyacrylamide gel electrophoresis with a strictly mass spectrometry-based approach for data analysis

被引:116
作者
Roessler, Markus
Rollinger, Wolfgang
Mantovani-Endl, Liliana
Hagmann, Marie-Luise
Palme, Stefan
Berndt, Peter
Engel, Alfred M.
Pfeffer, Michael
Karl, Johann
Bodenmueller, Heinz
Rueschoff, Josef
Henkel, Thomas
Rohr, Gerhard
Rossol, Siegbert
Roesch, Wolfgang
Langen, Hanno
Zolg, Werner
Tacke, Michael
机构
[1] Roche Diagnost GmbH, Centralized Diagnost, D-82377 Penzberg, Germany
[2] F Hoffmann La Roche & Co Ltd, Roche Ctr Med Genomics, CH-4070 Basel, Switzerland
[3] Klinikum Kassel GmbH, Inst Pathol & Biomed Res, D-34125 Kassel, Germany
[4] Hochtaunus Kliniken GGmbH, Inst Gastroenterol, D-61348 Bad Homburg Vor Der Hohe, Germany
[5] Stadtkrankenhaus Russelsheim, Med Abt, D-65428 Russelsheim, Germany
[6] Krankenhaus NW Frankfurt, D-60488 Frankfurt, Germany
关键词
D O I
10.1074/mcp.M600118-MCP200
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The purpose of this study was to identify and validate novel serological protein biomarkers of human colorectal cancer (CRC). Proteins from matched CRC and adjacent normal tissue samples were resolved by two-dimensional gel electrophoresis. From each gel all spots were excised, and enveloped proteins were identified by MS. By comparison of the resulting protein profiles, dysregulated proteins can be identified. A list of all identified proteins and validation of five exemplarily selected proteins, elevated in CRC was reported previously (Roessler, M., Rollinger, W., Palme, S., Hagmann, M. L., Berndt, P., Engel, A. M., Schneidinger, B., Pfeffer, M., Andres, H., Karl, J., Bodenmuller, H., Ruschoff, J., Henkel, T., Rohr, G., Rossol, S., Rosch, W., Langen, H., Zolg, W., and Tacke, M. ( 2005) Identification of nicotinamide N-methyltransferase as a novel serum tumor marker for colorectal cancer. Clin. Cancer Res. 11, 6550-6557). Here we describe identification and initial validation of another potential marker protein for CRC. Comparison of tissue protein profiles revealed strong elevation of proteasome activator complex subunit 3 (PSME3) expression in CRC tissue. This dysregulation was not detectable based on the spot pattern. The PSME3-containing spot on tumor gels showed no visible difference to the corresponding spot on matched control gels. MS analysis revealed the presence of two proteins, PSME3 and annexin 4 (ANXA4) in one and the same spot on tumor gels, whereas the matched spot contained only one protein, ANXA4, on control gels. Therefore, dysregulation of PSME3 was masked by ANXA4 and could only be recognized by MS-based analysis but not by image analysis. To validate this finding, antibody to PSME3 was developed, and up-regulation in CRC was confirmed by Western blot analysis and immunohistochemistry. Finally by developing a highly sensitive immunoassay, PSME3 could be detected in human sera and was significantly elevated in CRC patients compared with healthy donors and patients with benign bowel disease. We propose that PSME3 be considered a novel serum tumor marker for CRC that may have significance in the detection and in the management of patients with this disease. Further studies are needed to fully assess the potential clinical value of this marker candidate.
引用
收藏
页码:2092 / 2101
页数:10
相关论文
共 43 条
[1]   Fecal occult blood testing for colorectal cancer - Can we afford to do this? [J].
Ahlquist, DA .
GASTROENTEROLOGY CLINICS OF NORTH AMERICA, 1997, 26 (01) :41-&
[2]   Immune defects in 28-kDa proteasome activator γ-deficient mice [J].
Barton, LF ;
Runnels, HA ;
Schell, TD ;
Cho, YJ ;
Gibbons, R ;
Tevethia, SS ;
Deepe, GS ;
Monaco, JJ .
JOURNAL OF IMMUNOLOGY, 2004, 172 (06) :3948-3954
[3]  
Bast RC, 1996, J CLIN ONCOL, V14, P2843
[4]   2000 update of recommendations for the use of tumor markers in breast and colorectal cancer: Clinical practice guidelines of the American Society of Clinical Oncology [J].
Bast, RC ;
Ravdin, P ;
Hayes, DF ;
Bates, S ;
Fritsche, H ;
Jessup, JM ;
Kemeny, N ;
Locker, GY ;
Mennel, RG ;
Somerfield, MR .
JOURNAL OF CLINICAL ONCOLOGY, 2001, 19 (06) :1865-1878
[5]  
Berndt P, 1999, ELECTROPHORESIS, V20, P3521, DOI 10.1002/(SICI)1522-2683(19991201)20:18<3521::AID-ELPS3521>3.0.CO
[6]  
2-8
[7]   Spot overlapping in two-dimensional maps: A serious problem ignored for much too long [J].
Campostrini, N ;
Areces, LB ;
Rappsilber, J ;
Pietrogrande, MC ;
Dondi, F ;
Pastorino, F ;
Ponzoni, M ;
Righetti, PG .
PROTEOMICS, 2005, 5 (09) :2385-2395
[8]  
Ciechanover A, 2000, BIOESSAYS, V22, P442, DOI 10.1002/(SICI)1521-1878(200005)22:5<442::AID-BIES6>3.0.CO
[9]  
2-Q
[10]   Accuracy of screening for fecal occult blood on a single stool sample obtained by digital rectal examination: A comparison with recommended sampling practice [J].
Collins, JF ;
Lieberman, DA ;
Durbin, TE ;
Weiss, DG .
ANNALS OF INTERNAL MEDICINE, 2005, 142 (02) :81-85