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Detection of Melanoma Cells in the Lymphatic Drainage after Lymph Node Dissection in Melanoma Patients by Using Two-Marker Reverse Transcriptase-Polymerase Chain Reaction Assay

Ruka Wodzimierz, MD, Piotr Rutkowski, MD, Zbigniew I. Nowecki, MD, Jadwiga Kulik, MD, Anna Nasierowska-Guttmejer, MD and Janusz A. Siedlecki, PhD

From the Departments of Soft Tissue/Bone Sarcoma and Melanoma (RW, PR, ZIN), Molecular Biology (JK, JAS), and Pathology (AN-G), M. Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, Warsaw, Poland.

Correspondence: Address correspondence and reprint requests to: Piotr Rutkowski, MD, Department of Soft Tissue/Bone Sarcoma and Melanoma, M. Sklodowska-Curie Memorial Cancer Center and Institute of Oncology, W. Roentgena Str. 5, 02-781, Warsaw, Poland; Fax: 48-22-643-9791; E-mail: rutkowskip{at}coi.waw.pl

Background: The aim of this study was to evaluate the role of melanoma gene expression as a marker of the presence of melanoma cells in lymphatic drainage routinely collected after lymphadenectomy and to correlate reverse transcriptase-polymerase chain reaction (RT-PCR) assay results with recurrence, survival, and prognostic factors.

Methods: We collected 24-hour postoperative lymphatic drainage samples (between days 2 and 4) from 93 patients with stage III melanoma who underwent radical lymphadenectomy between May 2002 and November 2003. We used RT-PCR assays with primers specific for the tyrosinase and MART-1 (Melan-A) genes. The samples were considered positive if at least one marker was expressed. Median follow-up time was 12.8 months.

Results: In 18 (19.4%) of 93 patients, the RT-PCR assay results were positive: in 8 of 18 for tyrosinase only, in 7 of 18 for MART-1 only, and in 3 of 18 for both markers. We observed a significantly higher recurrence rate in patients with positive RT-PCR results (15 of 18; 83%) than negative results (26 of 75; 35%; P = .0001). Positive results of RT-PCR correlated with the number of involved lymph nodes (P = .0001) and extracapsular extension of nodal metastases (P = .03). We observed significant differences in overall and disease-free survival for RT-PCR–positive and –negative patients in univariate and multivariate analyses.

Conclusions: We observed positive RT-PCR assay results for melanoma cells in the lymphatic drainages of approximately 20% of patients after lymphadenectomy. This correlated significantly with early recurrence and shorter survival. These results may suggest that the RT-PCR assay could be useful for routinely analyzing postoperatively collected lymphatic drainage in stage III melanoma patients and for predicting disease progression.

Key Words: Melanoma • Lymph node • Metastasis • RT-PCR assay • Prognostic markers