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Proteomics Clin Appl. 2019 Jan;13(1):e1700173. doi: 10.1002/prca.201700173. Epub 2018 Nov 26.

Identification of Proteomic Markers in Head and Neck Cancer Using MALDI-MS Imaging, LC-MS/MS, and Immunohistochemistry.

Author information

1
Department of Otorhinolaryngology, Jena University Hospital, Jena, Germany.
2
Institute of Forensic Medicine, Section Pathology, Jena University Hospital, Jena, Germany.
3
Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Hans Knöll Institute, Jena, Germany.
4
Institute of Physical Chemistry, Friedrich Schiller University, Jena, Germany.

Abstract

PURPOSE:

The heterogeneity of squamous cell carcinoma tissue greatly complicates diagnosis and individualized therapy. Therefore, characterizing the heterogeneity of tissue spatially and identifying appropriate biomarkers is crucial. MALDI-MS imaging (MSI) is capable of analyzing spatially resolved tissue biopsies on a molecular level.

EXPERIMENTAL DESIGN:

MALDI-MSI is used on snap frozen and formalin-fixed and paraffin-embedded (FFPE) tissue samples from patients with head and neck cancer (HNC) to analyze m/z values localized in tumor and nontumor regions. Peptide identification is performed using LC-MS/MS and immunohistochemistry (IHC).

RESULTS:

In both FFPE and frozen tissue specimens, eight characteristic masses of the tumor's epithelial region are found. Using LC-MS/MS, the peaks are identified as vimentin, keratin type II, nucleolin, heat shock protein 90, prelamin-A/C, junction plakoglobin, and PGAM1. Lastly, vimentin, nucleolin, and PGAM1 are verified with IHC.

CONCLUSIONS AND CLINICAL RELEVANCE:

The combination of MALDI-MSI, LC-MS/MS, and subsequent IHC furnishes a tool suitable for characterizing the molecular heterogeneity of tissue. It is also suited for use in identifying new representative biomarkers to enable a more individualized therapy.

KEYWORDS:

FFPE; LC-MS/MS; MALDI MSI; head and neck squamous cell carcinoma; tumor marker

PMID:
30411850
DOI:
10.1002/prca.201700173
[Indexed for MEDLINE]

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