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Proteome Analysis of Human Pancreatic Ductal Adenocarcinoma Tissue Using Two-Dimensional Gel Electrophoresis and Tandem Mass Spectrometry for Identification of Disease-Related Proteins

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Abstract

A comparative proteomic approach has been used to identify and analyze proteins related to pancreatic cancer. Proteomes of eight pairs of clinical pancreatic ductal adenocarcinoma (PDAC) tissue samples and samples of normal adjacent tissue were obtained by two-dimensional gel electrophoresis (2DE). Comprehensive analysis of proteins was focused on total protein spots for which there were statistical differences between the two groups. Proteins were identified by peptide mass fingerprinting with tandem mass spectrometry (MS–MS). Western blotting and immunohistochemistry (IHC) were also performed to verify the expression of some candidate proteins. Thirty protein spots were identified, including proteases, antioxidant proteins, signal-transduction proteins, calcium-binding proteins, structural proteins, chaperones, and others. Western blotting and IHC confirmed up-regulated expression of two candidate proteins, nucleotide diphosphatase kinase (NDPK) and annexin II, in tumorous tissues. These results suggest that combination of 2DE with MS is an effective strategy for discovery of differently expressed proteins in PDAC which may be molecular markers for diagnosis or therapeutic targets.

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Abbreviations

PDAC:

Pancreatic ductal adenocarcinoma

NDPK:

Nucleotide diphosphatase kinase

IHC:

Immunohistochemistry

IAA:

Iodoacetamide

CHAPS:

3-[(3-Cholamidopropyl)dimethylammonio]-1-propane sulfonate

DTT:

Dithiothuretiol

ACN:

Acetonitrile

TFA:

Trifluoroacetic acid

CHCA:

α-Cyano-4-hydroxycinnamic acid

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Acknowledgements

We thank Su-Jun Li (Research Center for Proteome Analysis, Institute of Biochemistry and Cell Biology, Shanghai, P.R. China) and Qing-Ping Liu (Institutes of Biomedical Sciences, Fudan University, Shanghai, P.R. China) for technical assistance. This work was supported by National Natural Science Foundation of China, No. 30471693.

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Authors and Affiliations

  1. Department of Biliary-Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, P.R. China

    Rui Tian, Ren-Yi Qin, Zhi-Yong Du, Wei Xia & Cheng-Jian Shi

  2. Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, P.R. China

    Li-Ming Wei

  3. Department of Dermatology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, P.R. China

    Yan Li

  4. Department of Chemistry, Fudan University, Shanghai, 200433, P.R. China

    Hong Jin

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  1. Rui Tian
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Correspondence to Ren-Yi Qin.

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Rui Tian and Li-Ming Wei contributed equally to this work

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Tian, R., Wei, LM., Qin, RY. et al. Proteome Analysis of Human Pancreatic Ductal Adenocarcinoma Tissue Using Two-Dimensional Gel Electrophoresis and Tandem Mass Spectrometry for Identification of Disease-Related Proteins. Dig Dis Sci 53, 65–72 (2008). https://doi.org/10.1007/s10620-007-9823-3

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  • DOI: https://doi.org/10.1007/s10620-007-9823-3

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