title: Detection of melanoma cells in sentinel lymph nodes, bone marrow and peripheral blood by a reverse transcription-polymerase chain reaction assay in patients with primary cutaneous melanoma: association with Breslow's tumour thickness.
abstract: BACKGROUND
  Tyrosinase reverse transcription-polymerase chain reaction (RT-PCR) has been shown to be highly sensitive in detecting tumour cells in melanoma patients.
  
  
  OBJECTIVE
  To assess whether the detection of minimal residual disease by RT-PCR is improved by concomitant analysis of sentinel lymph nodes (SLNs), bone marrow (BM) and peripheral blood (PB) in patients with primary melanoma.
  
  
  METHODS
  Thirty-five SLNs, 41 BM samples and 26 PB specimens from 26 patients with primary cutaneous melanoma (tumour thickness > or = 0.75 mm) were examined by nested RT-PCR for tyrosinase and Melan-A. SLNs and BM samples were also analysed by histopathology. RT-PCR findings were related to tumour thickness of the primary melanoma.
  
  
  RESULTS
  Overall, melanoma cells were detected by RT-PCR in 13 of 26 patients (50%). Seven patients had positive RT-PCR results in their SLNs (27%), including all patients (n = 4) with histologically positive SLNs, two patients had positive findings in their BM exclusively detected by RT-PCR (8%) and six patients in PB (23%). The presence of tumour cells detected by RT-PCR in SLNs was not related to the presence of melanoma cells in BM and/or PB. The incidence of RT-PCR-positive SLNs was significantly associated with greater tumour thickness (P = 0.004). Both patients with positive RT-PCR findings in their BM had a large tumour thickness (> or = 2 mm). No association between positive RT-PCR findings in PB and greater tumour thickness was observed.
  
  
  CONCLUSIONS
  RT-PCR-positive SLNs were strongly associated with greater tumour thickness, underlining the prognostic significance of SLN positivity. Similar to certain epithelial malignancies, molecular investigation of the BM might provide complementary prognostic information in the early stages of melanoma. In contrast, no association between positive RT-PCR results in PB and increasing tumour thickness was found, implying that RT-PCR findings in PB are of doubtful clinical relevance in primary melanoma.
authors.affiliation: Department of Dermatology, Skin Cancer Programme, Eberhard-Karls-University, Liebermeister Str. 25, 72076 Tübingen, Germany. hans-juergen.blaheta@med.uni-tuebingen.de
authors.forename: H J
authors.initials: HJ
authors.lastname: Blaheta
authors.forename: T
authors.initials: T
authors.lastname: Paul
authors.forename: K
authors.initials: K
authors.lastname: Sotlar
authors.forename: E
authors.initials: E
authors.lastname: Maczey
authors.forename: B
authors.initials: B
authors.lastname: Schittek
authors.forename: A
authors.initials: A
authors.lastname: Paul
authors.forename: M
authors.initials: M
authors.lastname: Moehrle
authors.forename: H
authors.initials: H
authors.lastname: Breuninger
authors.forename: B
authors.initials: B
authors.lastname: Bueltmann
authors.forename: G
authors.initials: G
authors.lastname: Rassner
authors.forename: C
authors.initials: C
authors.lastname: Garbe
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor / Q000032:analysis; D001856:Bone Marrow Examination / Q000379:methods; D005260:Female; D006801:Humans; D008207:Lymphatic Metastasis; D058965:MART-1 Antigen; D008297:Male; D008545:Melanoma / Q000378:metabolism / Q000473:pathology* / Q000556:secondary; D008875:Middle Aged; D014442:Monophenol Monooxygenase / Q000032:analysis; D009363:Neoplasm Proteins / Q000032:analysis; D009367:Neoplasm Staging / Q000379:methods; D011379:Prognosis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D021701:Sentinel Lymph Node Biopsy / Q000379:methods*; D012878:Skin Neoplasms / Q000378:metabolism / Q000473:pathology*; D018709:Statistics, Nonparametric
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D014442:Monophenol Monooxygenase