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Robert P. Takes, MD; Robert J. Baatenburg de Jong, MD, PhD; Ed Schuuring, PhD; Jo Hermans, PhD; André A. Vis; Sergey V. Litvinov, PhD; J. Han J. M. van Krieken, MD, PhD

Arch Otolaryngol Head Neck Surg. 1997;123(4):412-419.

Abstract
Background
Regional metastasis is an important factor in the treatment and prognosis of patients with head and neck squamous cell carcinoma. Although in recent years imaging techniques have improved, it is still impossible to detect small metastatic deposits. Metastasis is mainly determined by properties of the primary tumor and its interaction with surrounding structures.

Objective
To identify markers that predict the presence of metastasis based on the features of the primary tumor.

Design
Correlation of the results of histological, immunohistochemical, and molecular biological analysis with clinical and histopathological data.

Materials and Methods
Several histological features and biological markers were examined in 31 laryngeal carcinomas. The following markers were selected on their putative role in the process of metastasis and were studied using immunohistochemical and/or Southern blot techniques: proliferating cell nuclear antigen (PCNA),p53, retinoblastoma tumor-suppressor gene (Rb), myc, bcl-2 (inhibitor of apoptosis), epidermal growth factor (EGF), EGF-receptor (EGFR), neu, nm23 (also known as NME1, putative metastasis suppressor), desmoplakin, neuron cell-adhesion molecule (N-CAM), epithelial cell-adhesion molecule (Ep-CAM), E-cadherin, cyclin D1 (CCND1), and EMS1.

Results
The presence of an inflammatory reaction surrounding the tumor (P=.07), eosinophilic infiltration (P=.16), positive immunostaining for Rb (P=.02), negative immunostaining for Ep-CAM (P=.13), and amplification of CCND1 and EMS1 (P=.05) correlated with nodal metastasis. The combination of an inflammatory reaction, eosinophilic infiltration, and staining for Rb and Ep-CAM resulted in a superior accuracy in assessing nodal metastasis.

Conclusions
These results indicate that it is possible to predict and exclude lymph node metastasis by studying the features of the primary tumor only. When these results are confirmed in a larger series, biological markers may be powerful diagnostic tools with great impact on clinical decision making.

Arch Otolaryngol Head Neck Surg. 1997;123:412-419

Author Affiliations
From the Departments of Otolaryngology (Drs Takes and Baatenburg de Jong), Pathology (Drs Schuuring, Litvinov, and van Krieken and Mr Vis), and Biostatistics (Dr Hermans), University Hospital Leiden, Leiden, the Netherlands.

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