Functional & Integrative Genomics

, Volume 11, Issue 2, pp 225–239 | Cite as

Mitochondrial proteomics analysis of tumorigenic and metastatic breast cancer markers

  • Yi-Wen Chen
  • Hsiu-Chuan Chou
  • Ping-Chiang Lyu
  • Hsien-Sheng Yin
  • Fang-Liang Huang
  • Wun-Shaing Wayne Chang
  • Chiao-Yuan Fan
  • I-Fan Tu
  • Tzu-Chia Lai
  • Szu-Ting Lin
  • Ying-Chieh Lu
  • Chieh-Lin Wu
  • Shun-Hong Huang
  • Hong-Lin Chan
Original Paper

Abstract

Mitochondria are key organelles in mammary cells responsible for several cellular functions including growth, division, and energy metabolism. In this study, mitochondrial proteins were enriched for proteomics analysis with the state-of-the-art two-dimensional differential gel electrophoresis and matrix-assistant laser desorption ionization–time-of-flight mass spectrometry strategy to compare and identify the mitochondrial protein profiling changes between three breast cell lines with different tumorigenicity and metastasis. The proteomics results demonstrate more than 1,500 protein features were resolved from the equal amount pooled from three purified mitochondrial proteins, and 125 differentially expressed spots were identified by their peptide finger print, in which, 33 identified proteins belonged to mitochondrial proteins. Eighteen out of these 33 identified mitochondrial proteins such as SCaMC-1 have not been reported in breast cancer research to our knowledge. Additionally, mitochondrial protein prohibitin has shown to be differentially distributed in mitochondria and in nucleus for normal breast cells and breast cancer cell lines, respectively. To sum up, our approach to identify the mitochondrial proteins in various stages of breast cancer progression and the identified proteins may be further evaluated as potential breast cancer markers in prognosis and therapy.

Keywords

Breast cancer Biomarker Proteomics Mitochondria DIGE MALDI-TOF Tumorigenesis Metastasis Prohibitin SCaMC-1 

Abbreviations

1-DE

One-dimensional gel electrophoresis

2-DE

Two-dimensional gel electrophoresis

Ab

Antibody

CCB

Colloidal Coomassie blue

CHAPS

3-[(3-Cholamidopropyl)-dimethylammonio]-1-propanesulfonate)

ddH2O

Double deionized water

DIGE

Differential gel electrophoresis

DTT

Dithiothreitol

FCS

Fetal calf serum

MALDI-TOF MS

Matrix-assisted laser desorption ionization–time-of-flight mass spectrometry

NP-40

Nonidet P-40

TFA

Trifluoroacetic acid

Notes

Acknowledgments

This work was supported by grant (NSC 99-2311-B-007-002) from the National Science Council, Taiwan, NTHU Booster grant (99N2908E1) from the National Tsing Hua University, and grant (VGHUST99-P5-22) Veteran General Hospitals University System of Taiwan.

Supplementary material

10142_2011_210_MOESM1_ESM.pdf (150 kb)
Supplementary table 1 Alphabetic list of identified differentially expressed mitochondrial proteins across MCF-10A, MCF-7, and MDA-MB-231 breast cells obtained after 2D-DIGE coupled with MALDI-TOF mass spectrometry analysis. aFunctional class of identified mitochondrial proteins were referred to Uniprot Website (http://www.uniprot.org/). bAverage ratio of differentially expressed (p < 0.05) proteins across MCF-7/MCF-10A, MDA-MB-231/MCF-10A, and MDA-MB-231/MCF-7 calculated considering three replica gels. cIdentified proteins which have not been reported in any cancer research are marked “A”, while identified proteins which have been reported in cancer research but not in breast cancer are marked “B”. (PDF 149 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Yi-Wen Chen
    • 1
  • Hsiu-Chuan Chou
    • 2
  • Ping-Chiang Lyu
    • 1
  • Hsien-Sheng Yin
    • 1
  • Fang-Liang Huang
    • 3
  • Wun-Shaing Wayne Chang
    • 4
  • Chiao-Yuan Fan
    • 1
  • I-Fan Tu
    • 1
  • Tzu-Chia Lai
    • 1
  • Szu-Ting Lin
    • 1
  • Ying-Chieh Lu
    • 1
  • Chieh-Lin Wu
    • 1
  • Shun-Hong Huang
    • 1
  • Hong-Lin Chan
    • 1
  1. 1.Institute of Bioinformatics and Structural Biology and Department of Medical SciencesNational Tsing Hua UniversityHsinchuTaiwan
  2. 2.Department of Applied ScienceNational Hsinchu University of EducationHsinchuTaiwan
  3. 3.Pediatric DepartmentTaichung Veterans General HospitalTaichungTaiwan
  4. 4.National Institute of Cancer ResearchNational Health Research InstitutesZhunan TownTaiwan

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