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  Vol. 130 No. 1, January 2004 TABLE OF CONTENTS
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Molecular Assay to Detect Metastatic Head and Neck Squamous Cell Carcinoma

Marta T. Becker, MD; Carol G. Shores, MD, PhD; Kathy K. Yu, MD; Wendell G. Yarbrough, MD

Arch Otolaryngol Head Neck Surg. 2004;130:21-27.

Background  The presence or absence of metastatic disease in cervical lymph nodes is the single most important determinant of therapy and prognosis for patients with head and neck squamous cell carcinoma (HNSCC). However, histologic examination fails to detect metastatic disease in a subset of neck dissection specimens. The accuracy of neck staging may be improved by the use of molecular techniques. Cytokeratins 5, 14, and 20 may be appropriate markers for HNSCC because they are expressed in HNSCC but not in lymphatic tissue.

Design  To test the sensitivity of detection of cytokeratin 5, 14, and 20 messenger RNA by quantitative reverse transcription polymerase chain reaction (RT-PCR), full-length coding DNA sequences were cloned and transcribed. The expression of cytokeratin 5, 14, and 20 messenger RNA was quantified in 4 HNSCC cell lines and 11 tumors. A cell culture lymph node model was created.

Results  As few as 32 molecules of cytokeratin 14 could be detected using quantitative RT-PCR. Cytokeratins 5 and 14 were easily detected in all 4 HNSCC cell lines and almost all tumors. Cytokeratin 20 was not a useful marker, as expression was absent or significantly reduced in cell lines and tumors. In the lymph node model, cytokeratin 14 quantitative RT-PCR was able to detect 1 cancer cell in a background of 10 million lymphatic cells.

Conclusions  Quantitative RT-PCR detection of cytokeratin 5 or 14 is a sensitive new molecular technique that may be used for detection of cervical micrometastases in head and neck cancer.


From the Department of Otolaryngology/Head and Surgery (Drs Becker, Shores, and Yu) and Lineberger Comprehensive Cancer Center (Dr Shores), University of North Carolina at Chapel Hill; and Departments of Otolaryngology and Cancer Biology, Vanderbilt-Ingram Comprehensive Cancer Center, Vanderbilt University Medical Center, Nashville, Tenn (Dr Yarbrough). The authors have no relevant financial interest in this article.



THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES

The Diagnostic Accuracy of Reverse Transcription-PCR Quantification of Cytokeratin mRNA in the Detection of Sentinel Lymph Node Invasion in Oral and Oropharyngeal Squamous Cell Carcinoma: A Comparison with Immunohistochemistry
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Clin. Cancer Res. 2006;12:2498-2505.
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Molecular Staging of Cervical Lymph Nodes in Squamous Cell Carcinoma of the Head and Neck
Ferris et al.
Cancer Res. 2005;65:2147-2156.
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Clinical Evaluation of a New Molecular Method for Detection of Micrometastases in Head and Neck Squamous Cell Carcinoma
Shores et al.
Arch Otolaryngol Head Neck Surg 2004;130:937-942.
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