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

University of Dundee, Dundee, Scotland, United Kingdom
Clinical Cancer Research (Impact Factor: 8.72). 05/2006; 12(8):2498-505. DOI: 10.1158/1078-0432.CCR-05-2136
Source: PubMed


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.
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.
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 mum. 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.
Quantitative RT-PCR for SLN staging in cN(0) patients with oral and oropharyngeal squamous cell carcinoma seems to be a promising approach.

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    • "In this context, real-time reverse transcriptase-PCR (RT – PCR) may be helpful because it is operator independent and, with some adaptation, can be automated and rapidly performed (Raja et al, 2002; Hamakawa et al, 2004). In a previous study, we demonstrated that cytokeratin (CK)17 mRNA quantification could be evaluated in SLNs by semiquantitative RT – PCR and that neck staging could be performed with relevant sensitivity and specificity when compared with SS- IHC staging (Garrel et al, 2006). However, minute micrometastases (MIs) sized o450 mm produced signals similar to those of negative SLN controls and were thus undiagnosed by CK17 mRNA level quantification, showing the limitation of this mRNA marker for the detection of metastases in SLNs. "
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