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. 2018 Aug 29;11(1):73.
doi: 10.1186/s12920-018-0382-6.

Differentially expressed proteins in positive versus negative HNSCC lymph nodes

Alessandra Vidotto  1 , Giovana M Polachini  1 , Marina de Paula-Silva  2 , Sonia M Oliani  2 , Tiago Henrique  1 , Rossana V M López  3 , Patrícia M Cury  4 , Fabio D Nunes  5 , José F Góis-Filho  6 , Marcos B de Carvalho  7 , Andréia M Leopoldino  8 , Eloiza H Tajara  9   10
Affiliations

Differentially expressed proteins in positive versus negative HNSCC lymph nodes

Alessandra Vidotto et al. BMC Med Genomics. .

Abstract

Background: Lymph node metastasis is one of the most important prognostic factors in head and neck squamous cell carcinomas (HNSCCs) and critical for delineating their treatment. However, clinical and histological criteria for the diagnosis of nodal status remain limited. In the present study, we aimed to characterize the proteomic profile of lymph node metastasis from HNSCC patients.

Methods: In the present study, we used one- and two-dimensional electrophoresis and mass spectrometry analysis to characterize the proteomic profile of lymph node metastasis from HNSCC.

Results: Comparison of metastatic and non-metastatic lymph nodes showed 52 differentially expressed proteins associated with neoplastic development and progression. The results reinforced the idea that tumors from different anatomical subsites have dissimilar behaviors, which may be influenced by micro-environmental factor including the lymphatic network. The expression pattern of heat shock proteins and glycolytic enzymes also suggested an effect of the lymph node environment in controlling tumor growth or in metabolic reprogramming of the metastatic cell. Our study, for the first time, provided direct evidence of annexin A1 overexpression in lymph node metastasis of head and neck cancer, adding information that may be useful for diagnosing aggressive disease.

Conclusions: In brief, this study contributed to our understanding of the metastatic phenotype of HNSCC and provided potential targets for diagnostic in this group of carcinomas.

Keywords: Head and neck carcinoma; Lymph node; Metastasis; Proteomics.

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Conflict of interest statement

Ethics approval and consent to participate

The study was conducted under approval of the National Committee on Ethics in Research/CONEP (reference number 1763/05, 18/05/2005) and all patients signed informed consent to participate.

Consent for publication

All patients gave written informed consent to publication of their case details, including clinical and pathology report data.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Partial 2-DE gel images of differentially expressed proteins in metastatic (N+) and non-metastatic (N0) lymph nodes of HNSCC. Anatomical subsites - C02 (tongue), floor of the mouth (C04) and larynx carcinomas (C32) - and protein symbols/names are provided to the right of each panel. Over and underexpressed proteins are indicated with arrows
Fig. 2
Fig. 2
Immunodetection of E-FABP by Western blot. Representative Western blot illustrating the E-FABP expression in tumor-free (N0) and positive (N+) lymph nodes. β-actin was used as an internal control. MW=PageRuler Prestained Protein Ladder
Fig. 3
Fig. 3
Immunohistochemical analysis of E-FABP expression in oral squamous cell carcinoma and non-tumoral (margin) samples. a Intense positivity of E-FABP in nucleus and cytoplasm of the basal and spinous layer of the normal epithelium, b reaching all epithelial layers. Immunolabeling intensity and proportion varied in tumor samples, with (c) expression in nests of well differentiated areas, d heterogeneous pattern with predominance of low intensity level in tumor cells; and also (e) moderate and (f) high intensity level of staining in nests. Scale bar indicates 50 μm
Fig. 4
Fig. 4
Immunohistochemical analysis of ANXA1 expression in lymph nodes from head and neck carcinomas. a Non-metastatic lymph node (N0) samples: constitutive expression of ANXA1 in the subcapsular sinus. b Metastatic lymph node (N+) samples: endogenous ANXA1 expression increased in the lymph node tissue and in the metastatic cells (arrows). c Negative control of reaction. Sections: 2 μm. Counterstain: Hematoxylin. d Densitometry of ANXA1. Values expressed as mean ± S.E.M. *** p < 0,001
See this image and copyright information in PMC

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