Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2006

Open Access 01-12-2006 | Research article

Proteomic analysis of nipple aspirate fluid from women with early-stage breast cancer using isotope-coded affinity tags and tandem mass spectrometry reveals differential expression of vitamin D binding protein

Authors: Timothy M Pawlik, David H Hawke, Yanna Liu, Savitri Krishnamurthy, Herbert Fritsche, Kelly K Hunt, Henry M Kuerer

Published in: BMC Cancer | Issue 1/2006

Login to get access

Abstract

Background

Isotope-coded affinity tag (ICAT) tandem mass spectrometry (MS) allows for qualitative and quantitative analysis of paired protein samples. We sought to determine whether ICAT technology could quantify and identify differential expression of tumor-specific proteins in nipple aspirate fluid (NAF) from the tumor-bearing and contralateral disease-free breasts of patients with unilateral early-stage breast cancer.

Methods

Paired NAF samples from 18 women with stage I or II unilateral invasive breast carcinoma and 4 healthy volunteers were analyzed using ICAT labeling, sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE), liquid chromatography, and MS. Proteins were identified by sequence database analysis. Western blot analysis of NAF from an independent sample set from 12 women (8 with early-stage breast cancer and 4 healthy volunteers) was also performed.

Results

353 peptides were identified from tandem mass spectra and matched to peptide sequences in the National Center for Biotechnology Information database. Equal numbers of peptides were up- versus down-regulated. Alpha2HS-glycoprotein [Heavy:Light (H:L) ratio 0.63] was underexpressed in NAF from tumor-bearing breasts, while lipophilin B (H:L ratio 1.42), beta-globin (H:L ratio 1.98), hemopexin (H:L ratio 1.73), and vitamin D-binding protein precursor (H:L ratio 1.82) were overexpressed. Western blot analysis of pooled samples of NAF from healthy volunteers versus NAF from women with breast cancer confirmed the overexpression of vitamin D-binding protein in tumor-bearing breasts.

Conclusion

ICAT tandem MS was able to identify and quantify differences in specific protein expression between NAF samples from tumor-bearing and disease-free breasts. Proteomic screening techniques using ICAT and NAF may be used to find markers for diagnosis of breast cancer.
Appendix
Available only for authorised users
Literature
1.
Elmore JG, Barton MB, Moceri VM, Polk S, Arena PJ, Fletcher SW: Ten-year risk of false positive screening mammograms and clinical breast examinations. N Engl J Med. 1998, 338: 1089-1096.CrossRefPubMed
2.
3.
Petricoin EF, Ardekani AM, Hitt BA, Levine PJ, Fusaro VA, Steinberg SM, Mills GB, Simone C, Fishman DA, Kohn EC, Liotta LA: Use of proteomic patterns in serum to identify ovarian cancer. Lancet. 2002, 359: 572-577.CrossRefPubMed
4.
Wulfkuhle JD, Sgroi DC, Krutzsch H, McLean K, McGarvey K, Knowlton M, Chen S, Shu H, Sahin A, Kurek R, Wallwiener D, Merino MJ, Petricoin EF, Zhao Y, Steeg PS: Proteomics of human breast ductal carcinoma in situ. Cancer Res. 2002, 62: 6740-6749.PubMed
5.
Somiari RI, Sullivan A, Russell S, Somiari S, Hu H, Jordan R, George A, Katenhusen R, Buchowiecka A, Arciero C, Brzeski H, Hooke J, Shriver C: High-throughput proteomic analysis of human infiltrating ductal carcinoma of the breast. Proteomics. 2003, 3: 1863-1873.CrossRefPubMed
6.
Celis JE, Moreira JM, Gromova I, Cabezon T, Ralfkiaer U, Guldberg P, Straten PT, Mouridsen H, Friis E, Holm D, Rank F, Gromov P: Towards discovery-driven translational research in breast cancer. Febs J. 2005, 272: 2-15.CrossRefPubMed
7.
Arciero C, Somiari SB, Shriver CD, Brzeski H, Jordan R, Hu H, Ellsworth DL, Somiari RI: Functional relationship and gene ontology classification of breast cancer biomarkers. Int J Biol Markers. 2003, 18: 241-272.PubMed
8.
Jeffrey SS, Pollack JR: The diagnosis and management of pre-invasive breast disease: promise of new technologies in understanding pre-invasive breast lesions. Breast Cancer Res. 2003, 5: 320-328.CrossRefPubMedPubMedCentral
9.
Somiari RI, Somiari S, Russell S, Shriver CD: Proteomics of breast carcinoma. J Chromatogr B Analyt Technol Biomed Life Sci. 2005, 815: 215-225.CrossRefPubMed
10.
Zhang DH, Tai LK, Wong LL, Sethi SK, Koay ES: Proteomics of breast cancer: enhanced expression of cytokeratin19 in human epidermal growth factor receptor type 2 positive breast tumors. Proteomics. 2005, 5: 1797-1805.CrossRefPubMed
11.
Luo Y, Zhang J, Liu Y, Shaw AC, Wang X, Wu S, Zeng X, Chen J, Gao Y, Zheng D: Comparative proteome analysis of breast cancer and normal breast. Mol Biotechnol. 2005, 29: 233-244.CrossRefPubMed
12.
Bottari P, Aebersold R, Turecek F, Gelb MH: Design and synthesis of visible isotope-coded affinity tags for the absolute quantification of specific proteins in complex mixtures. Bioconjug Chem. 2004, 15: 380-388.CrossRefPubMed
13.
Paweletz CP, Trock B, Pennanen M, Tsangaris T, Magnant C, Liotta LA, Petricoin EF: Proteomic patterns of nipple aspirate fluids obtained by SELDI-TOF: potential for new biomarkers to aid in the diagnosis of breast cancer. Dis Markers. 2001, 17: 301-307.CrossRefPubMed
14.
Sauter ER, Shan S, Hewett JE, Speckman P, Du Bois GC: Proteomic analysis of nipple aspirate fluid using SELDI-TOF-MS. Int J Cancer. 2005, 114: 791-796.CrossRefPubMed
15.
Pawlik TM, Fritsche H, Coombes KR, Xiao L, Krishnamurthy S, Hunt KK, Pusztai L, Chen JN, Clarke CH, Arun B, Hung MC, Kuerer HM: Significant differences in nipple aspirate fluid protein expression between healthy women and those with breast cancer demonstrated by time-of-flight mass spectrometry. Breast Cancer Res Treat. 2005, 89: 149-157.CrossRefPubMed
16.
Ranish JA, Yi EC, Leslie DM, Purvine SO, Goodlett DR, Eng J, Aebersold R: The study of macromolecular complexes by quantitative proteomics. Nat Genet. 2003, 33: 349-355.CrossRefPubMed
17.
Zhou H, Ranish JA, Watts JD, Aebersold R: Quantitative proteome analysis by solid-phase isotope tagging and mass spectrometry. Nat Biotechnol. 2002, 20: 512-515.CrossRefPubMed
18.
Oda Y, Owa T, Sato T, Boucher B, Daniels S, Yamanaka H, Shinohara Y, Yokoi A, Kuromitsu J, Nagasu T: Quantitative chemical proteomics for identifying candidate drug targets. Anal Chem. 2003, 75: 2159-2165.CrossRefPubMed
19.
Gygi SP, Rist B, Gerber SA, Turecek F, Gelb MH, Aebersold R: Quantitative analysis of complex protein mixtures using isotope-coded affinity tags. Nat Biotechnol. 1999, 17: 994-999.CrossRefPubMed
20.
21.
Mo W, Karger BL: Analytical aspects of mass spectrometry and proteomics. Curr Opin Chem Biol. 2002, 6: 666-675.CrossRefPubMed
22.
Turecek F: Mass spectrometry in coupling with affinity capture-release and isotope-coded affinity tags for quantitative protein analysis. J Mass Spectrom. 2002, 37: 1-14.CrossRefPubMed
23.
Link AJ, Eng J, Schieltz DM, Carmack E, Mize GJ, Morris DR, Garvik BM, Yates JR: Direct analysis of protein complexes using mass spectrometry. Nat Biotechnol. 1999, 17: 676-682.CrossRefPubMed
24.
Opiteck GJ, Lewis KC, Jorgenson JW, Anderegg RJ: Comprehensive on-line LC/LC/MS of proteins. Anal Chem. 1997, 69: 1518-1524.CrossRefPubMed
25.
Shiio Y, Eisenman RN, Yi EC, Donohoe S, Goodlett DR, Aebersold R: Quantitative proteomic analysis of chromatin-associated factors. J Am Soc Mass Spectrom. 2003, 14: 696-703.CrossRefPubMed
26.
Han DK, Eng J, Zhou H, Aebersold R: Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags and mass spectrometry. Nat Biotechnol. 2001, 19: 946-951.CrossRefPubMedPubMedCentral
27.
Patterson SD: Proteomics: the industrialization of protein chemistry. Curr Opin Biotechnol. 2000, 11: 413-418.CrossRefPubMed
28.
Hardwidge PR, Rodriguez-Escudero I, Goode D, Donohoe S, Eng J, Goodlett DR, Aebersold R, Finlay BB: Proteomic analysis of the intestinal epithelial cell response to enteropathogenic Escherichia coli. J Biol Chem. 2004, 279: 20127-20136.CrossRefPubMed
29.
Sartorius OW, Smith HS, Morris P, Benedict D, Friesen L: Cytologic evaluation of breast fluid in the detection of breast disease. J Natl Cancer Inst. 1977, 59: 1073-1080.PubMed
30.
Li J, Steen H, Gygi SP: Protein Profiling with Cleavable Isotope-coded Affinity Tag (cICAT) Reagents: The Yeast Salinity Stress Response. Mol Cell Proteomics. 2003, 2: 1198-1204.CrossRefPubMed
31.
Freake HC, Abeyasekera G, Iwasaki J, Marcocci C, MacIntyre I, McClelland RA, Skilton RA, Easton DF, Coombes RC: Measurement of 1,25-dihydroxyvitamin D3 receptors in breast cancer and their relationship to biochemical and clinical indices. Cancer Res. 1984, 44: 1677-1681.PubMed
32.
Eisman JA, Suva LJ, Sher E, Pearce PJ, Funder JW, Martin TJ: Frequency of 1,25-dihydroxyvitamin D3 receptor in human breast cancer. Cancer Res. 1981, 41: 5121-5124.PubMed
33.
Findlay DM, Michelangeli VP, Eisman JA, Frampton RJ, Moseley JM, MacIntyre I, Whitehead R, Martin TJ: Calcitonin and 1,25-dihydroxyvitamin D3 receptors in human breast cancer cell lines. Cancer Res. 1980, 40: 4764-4767.PubMed
34.
Eisman JA, Martin TJ, MacIntyre I, Moseley JM: 1,25-dihydroxyvitamin-D-receptor in breast cancer cells. Lancet. 1979, 2: 1335-1336.CrossRefPubMed
35.
Sauter ER, Ross E, Daly M, Klein-Szanto A, Engstrom PF, Sorling A, Malick J, Ehya H: Nipple aspirate fluid: a promising non-invasive method to identify cellular markers of breast cancer risk. Br J Cancer. 1997, 76: 494-501.CrossRefPubMedPubMedCentral
36.
Klein P, Glaser E, Grogan L, Keane M, Lipkowitz S, Soballe P, Brooks L, Jenkins J, Steinberg SM, DeMarini DM, Kirsch I: Biomarker assays in nipple aspirate fluid. Breast J. 2001, 7: 378-387.CrossRefPubMed
37.
Kuerer HM, Goldknopf IL, Fritsche H, Krishnamurthy S, Sheta EA, Hunt KK: Identification of distinct protein expression patterns in bilateral matched pair breast ductal fluid specimens from women with unilateral invasive breast carcinoma. High-throughput biomarker discovery. Cancer. 2002, 95: 2276-2282.CrossRefPubMed
38.
Sauter ER, Daly M, Linahan K, Ehya H, Engstrom PF, Bonney G, Ross EA, Yu H, Diamandis E: Prostate-specific antigen levels in nipple aspirate fluid correlate with breast cancer risk. Cancer Epidemiol Biomarkers Prev. 1996, 5: 967-970.PubMed
39.
Liu Y, Wang JL, Chang H, Barsky SH, Nguyen M: Breast-cancer diagnosis with nipple fluid bFGF. Lancet. 2000, 356: 567-CrossRefPubMed
40.
Zhao Y, Verselis SJ, Klar N, Sadowsky NL, Kaelin CM, Smith B, Foretova L, Li FP: Nipple fluid carcinoembryonic antigen and prostate-specific antigen in cancer-bearing and tumor-free breasts. J Clin Oncol. 2001, 19: 1462-1467.PubMed
41.
42.
Swallow CJ, Partridge EA, Macmillan JC, Tajirian T, DiGuglielmo GM, Hay K, Szweras M, Jahnen-Dechent W, Wrana JL, Redston M, Gallinger S, Dennis JW: alpha2HS-glycoprotein, an antagonist of transforming growth factor beta in vivo, inhibits intestinal tumor progression. Cancer Res. 2004, 64: 6402-6409.CrossRefPubMed
43.
Colpitts TL, Billing P, Granados E, Hodges S, Menhart N, Russell J, Stroupe S: Mammaglobin complexes with BU101 in breast tissue. Ann N Y Acad Sci. 2000, 923: 312-315.CrossRefPubMed
44.
Carter D, Douglass JF, Cornellison CD, Retter MW, Johnson JC, Bennington AA, Fleming TP, Reed SG, Houghton RL, Diamond DL, Vedvick TS: Purification and characterization of the mammaglobin/lipophilin B complex, a promising diagnostic marker for breast cancer. Biochemistry. 2002, 41: 6714-6722.CrossRefPubMed
45.
Carter D, Dillon DC, Reynolds LD, Retter MW, Fanger G, Molesh DA, Sleath PR, McNeill PD, Vedvick TS, Reed SG, Persing DH, Houghton RL: Serum antibodies to lipophilin B detected in late stage breast cancer patients. Clin Cancer Res. 2003, 9: 749-754.PubMed
46.
Fu SW, Schwartz A, Stevenson H, Pinzone JJ, Davenport GJ, Orenstein JM, Gutierrez P, Simmens SJ, Abraham J, Poola I, Stephan DA, Berg PE: Correlation of expression of BP1, a homeobox gene, with estrogen receptor status in breast cancer. Breast Cancer Res. 2003, 5: R82-7.CrossRefPubMedPubMedCentral
47.
Cui JF, Liu YK, Pan BS, Song HY, Zhang Y, Sun RX, Chen J, Feng JT, Tang ZY, Yu YL, Shen HL, Yang PY: Differential proteomic analysis of human hepatocellular carcinoma cell line metastasis-associated proteins. J Cancer Res Clin Oncol. 2004, 130: 615-622.CrossRefPubMed
48.
Yu B, Li SY, An P, Zhang YN, Liang ZJ, Yuan SJ, Cai HY: Comparative study of proteome between primary cancer and hepatic metastatic tumor in colorectal cancer. World J Gastroenterol. 2004, 10: 2652-2656.CrossRefPubMedPubMedCentral
49.
Thompson EW, Yu M, Bueno J, Jin L, Maiti SN, Palao-Marco FL, Pulyaeva H, Tamborlane JW, Tirgari R, Wapnir I, et al: Collagen induced MMP-2 activation in human breast cancer. Breast Cancer Res Treat. 1994, 31: 357-370.CrossRefPubMed
50.
Cao J, Kozarekar P, Pavlaki M, Chiarelli C, Bahou WF, Zucker S: Distinct roles for the catalytic and hemopexin domains of membrane type 1-matrix metalloproteinase in substrate degradation and cell migration. J Biol Chem. 2004, 279: 14129-14139.CrossRefPubMed
51.
Itoh Y, Takamura A, Ito N, Maru Y, Sato H, Suenaga N, Aoki T, Seiki M: Homophilic complex formation of MT1-MMP facilitates proMMP-2 activation on the cell surface and promotes tumor cell invasion. Embo J. 2001, 20: 4782-4793.CrossRefPubMedPubMedCentral
52.
Freake HC, Marcocci C, Iwasaki J, MacIntyre I: 1,25-dihydroxyvitamin D3 specifically binds to a human breast cancer cell line (T47D) and stimulates growth. Biochem Biophys Res Commun. 1981, 101: 1131-1138.CrossRefPubMed
53.
Dunkley TP, Dupree P, Watson RB, Lilley KS: The use of isotope-coded affinity tags (ICAT) to study organelle proteomes in Arabidopsis thaliana. Biochem Soc Trans. 2004, 32: 520-523.CrossRefPubMed
54.
Tian Q, Stepaniants SB, Mao M, Weng L, Feetham MC, Doyle MJ, Yi EC, Dai H, Thorsson V, Eng J, Goodlett D, Berger JP, Gunter B, Linseley PS, Stoughton RB, Aebersold R, Collins SJ, Hanlon WA, Hood LE: Integrated genomic and proteomic analyses of gene expression in Mammalian cells. Mol Cell Proteomics. 2004, 3: 960-969.CrossRefPubMed
55.
Yan W, Lee H, Yi EC, Reiss D, Shannon P, Kwieciszewski BK, Coito C, Li XJ, Keller A, Eng J, Galitski T, Goodlett DR, Aebersold R, Katze MG: System-based proteomic analysis of the interferon response in human liver cells. Genome Biol. 2004, 5: R54-CrossRefPubMedPubMedCentral
56.
Zang L, Palmer Toy D, Hancock WS, Sgroi DC, Karger BL: Proteomic analysis of ductal carcinoma of the breast using laser capture microdissection, LC-MS, and 16O/18O isotopic labeling. J Proteome Res. 2004, 3: 604-612.CrossRefPubMed
57.
Griffin TJ, Gygi SP, Rist B, Aebersold R, Loboda A, Jilkine A, Ens W, Standing KG: Quantitative proteomic analysis using a MALDI quadrupole time-of-flight mass spectrometer. Anal Chem. 2001, 73: 978-986.CrossRefPubMed
58.
Griffin TJ, Han DK, Gygi SP, Rist B, Lee H, Aebersold R, Parker KC: Toward a high-throughput approach to quantitative proteomic analysis: expression-dependent protein identification by mass spectrometry. J Am Soc Mass Spectrom. 2001, 12: 1238-1246.CrossRefPubMed
59.
Shiio Y, Donohoe S, Yi EC, Goodlett DR, Aebersold R, Eisenman RN: Quantitative proteomic analysis of Myc oncoprotein function. Embo J. 2002, 21: 5088-5096.CrossRefPubMedPubMedCentral
Metadata
Title
Proteomic analysis of nipple aspirate fluid from women with early-stage breast cancer using isotope-coded affinity tags and tandem mass spectrometry reveals differential expression of vitamin D binding protein
Authors
Timothy M Pawlik
David H Hawke
Yanna Liu
Savitri Krishnamurthy
Herbert Fritsche
Kelly K Hunt
Henry M Kuerer
Publication date
01-12-2006
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2006
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-6-68

Other articles of this Issue 1/2006

BMC Cancer 1/2006 Go to the issue

Research article

The relation between deoxycytidine kinase activity and the radiosensitising effect of gemcitabine in eight different human tumour cell lines

Study protocol

Prevalence of the use of cancer related self-tests by members of the public: a community survey

Research article

Trends in incidence and mortality of nasopharyngeal carcinoma over a 20–25 year period (1978/1983–2002) in Sihui and Cangwu counties in southern China

Research article

Prognostic significance of MCM2, Ki-67 and gelsolin in non-small cell lung cancer

Research article

Over-representation of specific regions of chromosome 22 in cells from human glioma correlate with resistance to 1,3-bis(2-chloroethyl)-1-nitrosourea

Research article

Inhibition of cyclooxygenase-2 decreases breast cancer cell motility, invasion and matrix metalloproteinase expression
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits