Melanoma patient staging: histopathological versus molecular evaluation of the sentinel node

Abstract

Lymphatic mapping and sentinel lymphadenectomy provide a minimally invasive means of directly determining the status of the regional lymph nodes in all patients who have a primary melanoma >1 mm thick but no clinical evidence of nodal involvement. Since the histological status of the sentinel node (SN) has been shown to be the most important prognostic factor in primary melanoma patients, the World Health Organization has recently recommended that sentinel lymphadenectomy should become the new standard of care for primary melanoma patients. This paper reviews the literature with regards to developments in and the current status of SN evaluation. Developments in the histopathological versus molecular detection of melanoma nodal metastases are reviewed, with specific emphasis on the strengths, limitations and clinical significance of these techniques. Molecular evaluation of the SN offers several advantages over standard histopathological analysis. These include an improved sensitivity, the cost-effective use of multiple markers for the improvement of detection rate and prognosis, as well as being less labour-intensive and costly. Moreover, molecular analysis has the potential to allow estimation of tumour burden. We review the potential causes of technical false-negative and false-positive reverse transcription-polymerase chain reaction (RT-PCR) results and how these could be eliminated by a systematic approach consisting of (i) careful and systematic assay design, which would include efficient tissue homogenization, choice of reagents and molecular markers, primer design and the use of one-stage versus two-stage PCR; (ii) careful optimization of the RT-PCR parameters (in particular the PCR cycle number) through the use of appropriate control tissues; and (iii) aiming for high assay reproducibility and lastly by applying the necessary positive and negative controls with each batch of samples. We also review the significant improvement in patient prognosis and management that has been made possible by the development of sentinel lymphadenectomy and histopathological evaluation of the SN, and compare the clinical (predictive) value of histopathological analysis with that of RT-PCR. Although RT-PCR is able to detect additional, clinically significant SN metastases that are missed by routine histopathology, its current limitation is that it overestimates the number of patients who have clinically significant melanoma metastases. Therefore, we suggest and discuss appropriate steps that need to be taken in order to minimize these false-positives and make this molecular tool more acceptable for routine clinical use.