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Untitled Document
CANCER GENOMICS & PROTEOMICS
Volume 2, Number
1, January-February 2005
| CONTENTS |
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| Genomics, Proteomics and Cancer: Specific Ribosomal, Mitochondrial, and Tumor Reactive Proteins Can Be Used as Biomarkers for Early Detection of Breast Cancer in Serum. J. ALBERTO FERNANDEZ-POL1, PAUL D. HAMILTON, DENNIS J. KLOS (Chesterfield, MO,U.S.A.) |
1 |
| A Display Thiol-Proteomics Approach to Characterize Global Redox Modification of Proteins by Selenium: Implications for the Anticancer Action of Selenium. E-M. PARK, K-S. CHOI, S-Y. PARK, E-S. KONG, K. ZU, Y. WU, H. O. ZHANG, C. IP, Y-M. PARK (Buffalo, NY,U.S.A.; Incheon,Korea) |
25 |
| *Protein Microarrays - A Promising Tool for Cancer Diagnosis. N. SCHNEIDERHAN-MARRA, A. KIRN, A. DOTTINGER, M. TEMPLIN, G. SAUER, H. DEISSLER, T. O. JOOS (Ulm; Reutlingen, Germany) |
37 |
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Detection of Specific mRNAs in Culture Medium Conditioned by Human Tumour Cells: Potential for New Class of Cancer Biomarkers in Serum. L. O'DRISCOLL, E. KENNY, M. PEREZ DE VILLARREAL, M. CLYNES (Dublin, Ireland) |
43 |
| Metallopanstimulin / S27 Ribosomal Antigen Expression in Stages I and II Breast Cancer: its Relationship with Clinicopathologic Factors. A. S. SUNDBLAD, L. RICCI, F. P. XYNOS, J. ALBERTO FERNANDEZ-POL (Chesterfield, Missouri, U.S.A.) |
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| *Review
(page 37) |
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CANCER GENOMICS & PROTEOMICS 2: 1-24 (2005)
Genomics, Proteomics and Cancer: Specific Ribosomal, Mitochondrial, and Tumor Reactive Proteins Can Be Used as Biomarkers for Early Detection of Breast Cancer in Serum
J. ALBERTO FERNANDEZ-POL1,2,3, PAUL D. HAMILTON1, DENNIS J. KLOS1
1Laboratory of Molecular Oncology, Department of Veterans Affairs Medical Center, St. Louis, MO 63106,
2Institute of Applied Neurociences, University of Buenos Aires, Facultad de Medicina, BsAs, Argentina, and
3Metalloproteomics, LLC, Chesterfield, MO 63017, U.S.A.
Abstract: We have used genomics and proteomics based technologies to study tissue and serum protein profiles in patients with breast cancer (BC) in comparison to control healthy subjects. One critical objective of this study was to identify and characterize new tissue and serum biomarkers of BC using differential screening of a recombinant cDNA human BC expression library. A second major objective of this study was to evaluate the clinical utility of Metallolpanstimulin (MPS-1/S27 ribosomal) protein as a biomarker for the early detection and monitoring of BC by immunoassay measurements of serum MPS-1 protein levels and to identify MPS-1 protein in sera of BC patients. A third objective was to present data on cloned BC genes denoted protein subgroup-30 (PS-30), consisting of mitochondria, nuclear, and ribosomal proteins which are increased after growth factor stimulation of BC cells in tissue culture. To study in detail MPS-1 protein expression in BC, the MPS-1 concentrations were determined in the blood of 117 females free of any disease, and in 203 female patients diagnosed with primary BC. The results indicate that increased serum MPS-1 levels can be used for the early detection of BC. Normal subjects have low concentrations of MPS-1 protein in sera. Moreover, changes in MPS-1 protein serum levels can be used for the study of BC progression or regression after various types of therapy. In both the low and high value range, MPS-1 is 10-fold more effective than CA-15-3 in modifying the probability of the target condition Ðbreast cancer. The use of HPLC, Western blot, Immuno-Mass Spectrometry, and protein sequencing confirmed the presence of authentic MPS-1 in sera of patients with BC. Negligible levels of MPS-1 protein were detected in sera from normal subjects. We conclude that (1) the increase in serum MPS-1 can be used for the early detection of BC; and (2) MPS-1 proved to be reliable in the follow-up of patients with advanced BC as demonstrated by the close correlation between MPS-1 protein levels and BC progression or regression after various types of therapy. Furthermore, all proteins denoted group -30 (Mr <30,000), consisting of ribosomal, nuclear and mitochondria proteins, were found to be significantly increased in BC tissues in comparison to control tissues, suggesting that these proteins may be useful markers for detection of BC. Finally, several serum reactive proteins such as haptoglobin and C3 complement components provided valuable information on oncogenic activity in BC patients.
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CANCER GENOMICS & PROTEOMICS 2: 53-60 (2005)
Metallopanstimulin / S27 Ribosomal AntigenExpression in Stages I and II Breast Cancer: its Relationship with Clinicopathologic Factors
ALBERTO S. SUNDBLAD1, LILA RICCI2, FRANCISCO P. XYNOS3, J. ALBERTO FERNANDEZ-POL4
1Departments of Pathology and 3Gynecologic Oncology, Division of Breast Cancer Screening/Surgery, Hospital Privado de Comunidad, Mar del Plata, Argentina, and
2Biostatistics Department, Facultad de Ciencias Exactas, National University of Mar del Plata,
Mar del Plata, Argentina, and
4Metalloproteomics, LLC, Chesterfield, Missouri 63017, U.S.A.
Abstract: Metallopanstimulin (MPS-1)/S27 ribosomal protein is involved in cellular proliferation and oncogenesis. In this study, we have examined the expression of the MPS-1 protein in 120 stages I and II breast carcinomas to study its relationship with breast cancer prognosis. We also determined if there was any relationship of MPS-1 with other biological markers commonly used in breast cancer prognosis. The expression of MPS-1 protein was analyzed by immunohistochemistry using specific anti-MPS-1 antibodies. We found that there was greater expression of MPS-1 in tumors of greater size and in higher histological grades. Thus, in tumors with more histological aggressiveness there is more MPS-1. Both were frequently associated with a greater proliferative activity. There was also a significant association between the expression of MPS-1 with the expression of receptors for progesterone (p=0.004), estrogens (p=0.03), bcl-2 (p=0.002), and MIB-1 (p=0.03). After univariate logistic regression analysis, we found that overexpression of MPS-1 correlated with Disease Free Survival (DFS) (p=0.039), showing that MPS-1 positivity is associated with a greater incidence of recurrence and/or metastasis. There was no association between overexpression of MPS-1 and poor Overall Survival (OS) (p=0.146). The results presented here indicate a significant correlation between overexpression of MPS-1/S27 ribosomal protein and more aggressive breast cancer growth. These results suggest that the MPS-1 antigen may be a useful marker to understand better the biological behavior of breast cancer.
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CANCER GENOMICS & PROTEOMICS 2: 37-42 (2005)
Protein Microarrays – A Promising Tool for Cancer Diagnosis
NICOLE SCHNEIDERHAN-MARRA1,2, ANETTE KIRN2, ANETTE DÏTTINGER2, MARKUS TEMPLIN2, GEORG SAUER1, HELMUT DEISSLER1, THOMAS O. JOOS2
1University of Ulm Medical School, Department of Obstetrics and Gynaecology, Frauensteige 14, 89075 Ulm;
2NMI Natural and Medical Sciences Institute at the University of Tubingen,Markwiesenstraâe 55, 72770 Reutlingen, Germany
Abstract: Within recent years, protein microarrays have been developed to quantify a large number of parameters present in a given sample simultaneously. Such miniaturised and parallelised sandwich immunoassays are of general interest for all proteomic and diagnostic approaches in which several parameters have to be determined from small samples, e.g. biopsy material. In addition to planar microarray-based approaches, bead-based flow cytometry is quite suitable for the multiplex detection of target molecules, especially when a limited number of parameters are to be analysed. Appropriate sensitivity, reproducibility and robustness have to be demonstrated before protein microarray technology can be used to characterise clinical samples and generate reliable data sets. As a model system to analyse these issues, a set of multiplexed sandwich immunoassays based on Luminex beads were developed to screen clinical samples for the presence or absence of marker proteins indicative of prognosis or response to therapeutic options.
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CANCER GENOMICS & PROTEOMICS 2: 25-36 (2005)
A Display Thiol-Proteomics Approach to Characterize Global Redox Modification of Proteins by Selenium: Implications for the Anticancer Action of Selenium
EUN-MI PARK1,3, KYOUNG-SOO CHOI1, SOO-YEON PARK1, EUNG-SIK KONG1, KE ZU2, YUE WU2, HAITAO ZHANG2, CLEMENT IP2, YOUNG-MEE PARK1
1Department of Cellular Stress Biology and 2Department of Cancer Chemoprevention, Roswell Park Cancer Institute, Elm & Carlton Streets, Buffalo, NY 14263, U.S.A.;
3Department of Chemistry, University of Incheon, Incheon, 402-749, Korea
Abstract: Background: The generation of a monomethylated selenium metabolite is critical for the anticancer activity of selenium. Because of its strong nucleophilicity, the metabolite can react directly with protein thiols to cause redox modification. These chemical changes have never been examined systematically before because of the lack of a reliable methodology to study reactive protein thiols globally in cells and to quantify their redox status. Materials and Methods: PC-3 human prostate cancer cells were treated with methylseleninic acid (MSA) for 0.5, 1, 2, 3, 6, 12 or 24 h. A reactive thiol specific reagent, BIAM, was used to detect the extent of global redox changes on a 2D gel electrophoresis display. The data were analyzed by the Self Organizing Maps clustering algorithm. Protein identification was done by MALDI-TOF and ESI-tandem mass spectrometry. Results: Out of a total of 194 reactive thiol-containing protein spots on the 2D gel display, 100 of them (cluster 1) were not sensitive to MSA modulation. The remaining 94 were categorized into three distinct patterns. Cluster 2 (60 proteins) showed an immediate and sustained loss of reactive thiols for at least 24 h; cluster 3 (19 proteins) showed a transient loss of reactive thiols followed by a rapid rebound; and cluster 4 (15 proteins) showed a transient gain followed by a rapid return to normal. In contrast, there were minimal protein redox changes in control cells (not treated with MSA) over the same period of time. A total of 85 proteins were identified of which 40 were in clusters 2 to 4. The proteins which are sensitive to redox modification by MSA are distributed in various subcellular compartments. Western blot analysis showed that a number of chaperones were significantly induced by MSA. Conclusion: Global redox modification of proteins can be a major driving force of cellular stress, since these changes are likely to lead to protein unfolding, misfolding or aggregation. The induction of chaperones in cells treated with MSA is consistent with this interpretation since chaperones are charged with rescuing misfolded proteins. The above scenario is discussed in relation to an adaptive response which ultimately determines how cells respond to treatment with selenium.
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CANCER GENOMICS & PROTEOMICS 2: 43-52 (2005)
Detection of Specific mRNAs in Culture Medium Conditioned by Human Tumour Cells: Potential for New Class of Cancer Biomarkers in Serum
LORRAINE O'DRISCOLL, ELAINE KENNY, MAIDER PEREZ DE VILLARREAL, MARTIN CLYNES
National Institute for Cellular Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland
Abstract: Background: This study aimed to develop and optimise procedures to enable us to establish, in a well-controlled environment, if cancer cells routinely secrete gene transcripts potentially suitable for monitoring as biomarkers. Materials and Methods: Aliquots of the conditioned media (CM) exposed to cancer cells (including breast, lung and nasal cancer cell lines) were removed at intervals of 24 hours over a 96-hour time period and were passed through 0.45 ìm or 0.22 ìm filters, to remove cellular material. Methods for subsequent RNA extraction (from CM and cells) were investigated. RT-PCR was performed for a number of mRNAs, including mdr-1, mrp-1, CK-19, HnRNP B1, GST-ð, topoisomerase II, bcl-2 and â-actin. Results: Gene transcripts, amplifiable by RT-PCR, were detected in conditioned media from all human cancer cell lines studied. This RNA did not result from the presence of cells in the conditioned media and, as expected, was absent from control medium not exposed to tumour cells. Conclusion: The results from this study indicate that gene transcripts may be secreted from human cancer cells and are detectable in the subsequent cell-free media. The methods developed and optimised here may be suitable for analysis of mRNAs as biomarkers in serum/plasma from cancer patients.
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