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

• Published in: Clinical cancer research Vol. 12; no. 8; pp. 2498 - 2505
• Main Authors: Garrel, Renaud, Dromard, Mathilde, Costes, Valérie, Barbotte, Eric, Comte, Frédéric, Gardiner, Quentin, Cartier, César, Makeieff, Marc, Crampette, Louis, Guerrier, Bernard, Boulle, Nathalie
• Format: Journal Article
• Language: English
• Published: United States American Association for Cancer Research 15.04.2006
• Subjects: Adult; Aged; Carcinoma, Squamous Cell - genetics; Carcinoma, Squamous Cell - metabolism; Carcinoma, Squamous Cell - pathology; diagnostic accuracy; head and neck cancer; Humans; Immunohistochemistry; Keratin-14; Keratin-5; Keratins - analysis; Keratins - genetics; Lymph Nodes - metabolism; Lymph Nodes - pathology; Lymphatic Metastasis - diagnosis; Lymphatic Metastasis - genetics; Middle Aged; Mouth Neoplasms - genetics; Mouth Neoplasms - metabolism; Mouth Neoplasms - pathology; Neoplasm Staging; Oropharyngeal Neoplasms - genetics; Oropharyngeal Neoplasms - metabolism; Oropharyngeal Neoplasms - pathology; quantitative RT-PCR; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sensitivity and Specificity; Sentinel Lymph Node Biopsy; sentinel lymph nodes
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.CCR-05-2136

Purpose: The main goal of sentinel lymph node (SLN) detection in head and neck squamous cell carcinomas is to limit neck dissections to pN+ cases only. However, intraoperative + diagnosis cannot be routinely done using the current gold standard, serial step sectioning with immunohistochemistry. Real-time quantitative reverse transcription-PCR (RT-PCR) is potentially compatible with intraoperative use, proving highly sensitive in detecting molecular markers. This study postoperatively assessed the accuracy of quantitative RT-PCR in staging patients from their SLN. Experimental Design: A combined analysis on the same SLN by serial step sectioning with immunohistochemistry and quantitative RT-PCR targeting cytokeratins 5, 14, and 17 was done in 18 consecutive patients with oral or oropharyngeal squamous cell carcinoma and 10 control subjects. Results: From 71 lymph nodes examined, mRNA levels (KRT) were linked to metastasis size for the three cytokeratins studied (Pearson correlation coefficient, r = 0.89, 0.73, and 0.77 for KRT 5, 14, and 17 respectively; P < 0.05). Histopathology-positive SLNs (macro- and micrometastases) showed higher mRNA values than negative SLNs for KRT 17 ( P < 10 −4 ) and KRT 14 ( P < 10 −2 ). KRT 5 showed nonsignificant results. KRT 17 seemed to be the most accurate marker for the diagnosis of micrometastases of a size >450 μm. Smaller micrometastases and isolated tumor cells did not provide results above the background level. Receiver operating characteristic curve analysis for KRT 17 identified a cutoff value where patient staging reached 100% specificity and sensitivity for macro- and micrometastases. Conclusion: Quantitative RT-PCR for SLN staging in cN 0 patients with oral and oropharyngeal squamous cell carcinoma seems to be a promising approach.