Identification of Stage-Specific Breast Markers Using Quantitative Proteomics

Sadr-ul Shaheed, Nitin Rustogi, Andrew Scally, Julie Wilson, Helene Thygesen, Maria A. Loizidou, Andreas Hadjisavvas, Andrew Hanby, Valerie Speirs, Paul Loadman, Richard Linforth, Kyriacos Kyriacou, Chris W. Sutton

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Matched healthy and diseased tissues from breast cancer patients were analyzed by quantitative proteomics. By comparing proteomic profiles of fibroadenoma (benign tumors, three patients), DCIS (noninvasive cancer, three patients), and invasive ductal carcinoma (four patients), we identified protein alterations that correlated with breast cancer progression. Three 8-plex iTRAQ experiments generated an average of 826 protein identifications, of which 402 were common. After excluding those originating from blood, 59 proteins were significantly changed in tumor compared with normal tissues, with the majority associated with invasive carcinomas. Bioinformatics analysis identified relationships between proteins in this subset including roles in redox regulation, lipid transport, protein folding, and proteasomal degradation, with a substantial number increased in expression due to Myc oncogene activation. Three target proteins, cofilin-1 and p23 (increased in invasive carcinoma) and membrane copper amine oxidase 3 (decreased in invasive carcinoma), were subjected to further validation. All three were observed in phenotype-specific breast cancer cell lines, normal (nontransformed) breast cell lines, and primary breast epithelial cells by Western blotting, but only cofilin-1 and p23 were detected by multiple reaction monitoring mass spectrometry analysis. All three proteins were detected by both analytical approaches in matched tissue biopsies emulating the response observed with proteomics analysis. Tissue microarray analysis (361 patients) indicated cofilin-1 staining positively correlating with tumor grade and p23 staining with ER positive status; both therefore merit further investigation as potential biomarkers.

Original languageEnglish
Pages (from-to)5696-5708
Number of pages13
JournalJournal of Proteome Research
Volume12
Issue number12
Early online date9 Oct 2013
DOIs
Publication statusPublished - 6 Dec 2013

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Cofilin 1
Proteomics
Breast
Tissue
Tumors
Breast Neoplasms
Proteins
Carcinoma
Neoplasms
Cells
Tissue Array Analysis
Staining and Labeling
Amine Oxidase (Copper-Containing)
Protein folding
Fibroadenoma
Cell Line
Ductal Carcinoma
myc Genes
Carcinoma, Intraductal, Noninfiltrating
Biopsy

Keywords

  • breast cancer
  • DCIS
  • fibroadenoma
  • invasive carcinoma
  • iTRAQ
  • mass spectrometry
  • proteomics
  • tissue microarray
  • Western blotting

Cite this

Shaheed, S., Rustogi, N., Scally, A., Wilson, J., Thygesen, H., Loizidou, M. A., ... Sutton, C. W. (2013). Identification of Stage-Specific Breast Markers Using Quantitative Proteomics. Journal of Proteome Research, 12(12), 5696-5708. https://doi.org/10.1021/pr400662k

Identification of Stage-Specific Breast Markers Using Quantitative Proteomics. / Shaheed, Sadr-ul; Rustogi, Nitin; Scally, Andrew; Wilson, Julie; Thygesen, Helene; Loizidou, Maria A.; Hadjisavvas, Andreas; Hanby, Andrew; Speirs, Valerie; Loadman, Paul; Linforth, Richard; Kyriacou, Kyriacos; Sutton, Chris W.

In: Journal of Proteome Research, Vol. 12, No. 12, 06.12.2013, p. 5696-5708.

Research output: Contribution to journalArticle

Shaheed, S, Rustogi, N, Scally, A, Wilson, J, Thygesen, H, Loizidou, MA, Hadjisavvas, A, Hanby, A, Speirs, V, Loadman, P, Linforth, R, Kyriacou, K & Sutton, CW 2013, 'Identification of Stage-Specific Breast Markers Using Quantitative Proteomics', Journal of Proteome Research, vol. 12, no. 12, pp. 5696-5708. https://doi.org/10.1021/pr400662k
Shaheed S, Rustogi N, Scally A, Wilson J, Thygesen H, Loizidou MA et al. Identification of Stage-Specific Breast Markers Using Quantitative Proteomics. Journal of Proteome Research. 2013 Dec 6;12(12):5696-5708. https://doi.org/10.1021/pr400662k
Shaheed, Sadr-ul ; Rustogi, Nitin ; Scally, Andrew ; Wilson, Julie ; Thygesen, Helene ; Loizidou, Maria A. ; Hadjisavvas, Andreas ; Hanby, Andrew ; Speirs, Valerie ; Loadman, Paul ; Linforth, Richard ; Kyriacou, Kyriacos ; Sutton, Chris W. / Identification of Stage-Specific Breast Markers Using Quantitative Proteomics. In: Journal of Proteome Research. 2013 ; Vol. 12, No. 12. pp. 5696-5708.
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abstract = "Matched healthy and diseased tissues from breast cancer patients were analyzed by quantitative proteomics. By comparing proteomic profiles of fibroadenoma (benign tumors, three patients), DCIS (noninvasive cancer, three patients), and invasive ductal carcinoma (four patients), we identified protein alterations that correlated with breast cancer progression. Three 8-plex iTRAQ experiments generated an average of 826 protein identifications, of which 402 were common. After excluding those originating from blood, 59 proteins were significantly changed in tumor compared with normal tissues, with the majority associated with invasive carcinomas. Bioinformatics analysis identified relationships between proteins in this subset including roles in redox regulation, lipid transport, protein folding, and proteasomal degradation, with a substantial number increased in expression due to Myc oncogene activation. Three target proteins, cofilin-1 and p23 (increased in invasive carcinoma) and membrane copper amine oxidase 3 (decreased in invasive carcinoma), were subjected to further validation. All three were observed in phenotype-specific breast cancer cell lines, normal (nontransformed) breast cell lines, and primary breast epithelial cells by Western blotting, but only cofilin-1 and p23 were detected by multiple reaction monitoring mass spectrometry analysis. All three proteins were detected by both analytical approaches in matched tissue biopsies emulating the response observed with proteomics analysis. Tissue microarray analysis (361 patients) indicated cofilin-1 staining positively correlating with tumor grade and p23 staining with ER positive status; both therefore merit further investigation as potential biomarkers.",
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