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S100-A10, thioredoxin, and S100-A6 as biomarkers of papillary thyroid carcinoma with lymph node metastasis identified by MALDI Imaging

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Abstract

In papillary thyroid carcinoma (PTC), metastasis is a feature of an aggressive tumor phenotype. To identify protein biomarkers that distinguish patients with an aggressive tumor behavior, proteomic signatures in metastatic and non-metastatic tumors were investigated comparatively. In particular, matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) was used to analyze primary tumor samples. We investigated a tumor cohort of PTC (n = 118) that were matched for age, tumor stage, and gender. Proteomic screening by MALDI-IMS was performed for a discovery set (n = 29). Proteins related to the discriminating mass peaks were identified by 1D-gel electrophoresis followed by mass spectrometry. The candidate proteins were subsequently validated by immunohistochemistry (IHC) using a tissue microarray for an independent PTC validation set (n = 89). In this study, we found 36 mass-to-charge-ratio (m/z) species that specifically distinguished metastatic from non-metastatic tumors, among which m/z 11,608 was identified as thioredoxin, m/z 11,184 as S100-A10, and m/z 10,094 as S100-A6. Furthermore, using IHC on the validation set, we showed that the overexpression of these three proteins was highly associated with lymph node metastasis in PTC (p < 0.005). For functional analysis of the metastasis-specific proteins, we performed an Ingenuity Pathway Analysis and discovered a strong relationship of all candidates with the TGF-β-dependent EMT pathway. Our results demonstrated the potential application of the MALDI-IMS proteomic approach in identifying protein markers of metastasis in PTC. The novel protein markers identified in this study may be used for risk stratification regarding metastatic potential in PTC.

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Acknowledgements

The authors gratefully acknowledge the financial support of the Federal Ministry of Education and Research (BMBF) Germany. The authors thank Ulrike Buchholz, Claudia-Mareike Pflueger, Andreas Voss, and Nina Weber for providing excellent technical assistance.

Conflict of Interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Department of Radiation Sciences, Research Unit of Radiation Cytogenetics, Helmholtz Zentrum München-German Research Center for Environmental Health GmbH, Ingolstaedter Landstraße 1, 85764, Neuherberg, Germany

    Martin Nipp & Horst Zitzelsberger

  2. Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environmental Health GmbH, Neuherberg, Germany

    Mareike Elsner, Benjamin Balluff, Stephan Meding, Sandra Rauser, Heinz Höfler & Axel Walch

  3. Department of Medicine II, Klinikum rechts der Isar, Technische Universität München, Munich, Germany

    Benjamin Balluff

  4. Research Unit of Protein Science, Helmholtz Zentrum München-German Research Center for Environmental Health GmbH, Neuherberg, Germany

    Hakan Sarioglu & Marius Ueffing

  5. Human Cancer Studies Group, Department of Surgery and Cancer, Hammersmith Hospital, Imperial College London, London, UK

    Kristian Unger

  6. Institute of Pathology, Technische Universität München, Munich, Germany

    Heinz Höfler

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  1. Martin Nipp
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Nipp, M., Elsner, M., Balluff, B. et al. S100-A10, thioredoxin, and S100-A6 as biomarkers of papillary thyroid carcinoma with lymph node metastasis identified by MALDI Imaging. J Mol Med 90, 163–174 (2012). https://doi.org/10.1007/s00109-011-0815-6

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