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Mitochondrial proteomics analysis of tumorigenic and metastatic breast cancer markers

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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.

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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

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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.

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Authors and Affiliations

  1. Institute of Bioinformatics and Structural Biology and Department of Medical Sciences, National Tsing Hua University, No. 101, Kuang-Fu Rd. Sec. 2, Hsinchu, 30013, Taiwan

    Yi-Wen Chen, Ping-Chiang Lyu, Hsien-Sheng Yin, Chiao-Yuan Fan, I-Fan Tu, Tzu-Chia Lai, Szu-Ting Lin, Ying-Chieh Lu, Chieh-Lin Wu, Shun-Hong Huang & Hong-Lin Chan

  2. Department of Applied Science, National Hsinchu University of Education, Hsinchu, Taiwan

    Hsiu-Chuan Chou

  3. Pediatric Department, Taichung Veterans General Hospital, Taichung, Taiwan

    Fang-Liang Huang

  4. National Institute of Cancer Research, National Health Research Institutes, Zhunan Town, Miaoli, 350, Taiwan

    Wun-Shaing Wayne Chang

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  1. Yi-Wen Chen
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  14. Hong-Lin Chan
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Correspondence to Hong-Lin Chan.

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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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Chen, YW., Chou, HC., Lyu, PC. et al. Mitochondrial proteomics analysis of tumorigenic and metastatic breast cancer markers. Funct Integr Genomics 11, 225–239 (2011). https://doi.org/10.1007/s10142-011-0210-y

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  • DOI: https://doi.org/10.1007/s10142-011-0210-y

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