Multimarker real-time reverse transcription-PCR for quantitative detection of melanoma-associated antigens: a novel possible staging method.
BACKGROUND
Detection of melanoma cells in peripheral blood is a promising method for monitoring haematogenous spread of melanoma cells. It enables us to detect early metastasis and to better stratify candidates for adjuvant immunotherapy. Inconsistent data on the sensitivity and clinical relevance of this method have been reported.


METHODS
We developed a multimarker real-time reverse transcription-PCR (RT-PCR) for quantification of five melanoma markers: Melan-A, gp100, MAGE-3, MIA and tyrosinase. In this prospective study, 65 patients with resected cutaneous melanoma stage IIB-III were screened. Peripheral blood samples were collected every 3 months for the following 18 months, and circulating melanoma cells were examined and compared with clinical staging results.


RESULTS
Eighteen patients relapsed during the trial and showed different types of melanoma progression. All these patients experienced statistically significant tumour marker elevation in the period from 0 to 9 months before the disease progression. MAGE-3 was the most sensitive progression marker. In patients with progression, we observed three concordant positive markers in 39% of cases, two concordant positive markers in 28%, and finally one marker in 33%.


CONCLUSIONS
This report describes a multiple-marker real-time RT-PCR, which is able to provide quantitative data on melanoma markers in the peripheral blood of melanoma patients. Measurement of the studied molecular markers in our hands represents a prognostic factor and a useful method for early detection of metastasis and treatment response of melanoma patients.
Arenberger
P
P
Department of Dermatology, Charles University 3rd Medical Faculty, Prague, Czech Republic.
Arenbergerova
M
M
Gkalpakiotis
S
S
Lippert
J
J
Stribrna
J
J
Kremen
J
J
D000328:Adult; D000368:Aged; D000951:Antigens, Neoplasm / Q000097:blood / Q000235:genetics; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000097:blood / Q000235:genetics; D014408:Biomarkers, Tumor / Q000097:blood*; D045744:Cell Line, Tumor; D018450:Disease Progression; D005260:Female; D006801:Humans; D058965:MART-1 Antigen; D008297:Male; D008545:Melanoma / Q000097:blood* / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000097:blood / Q000235:genetics; D008875:Middle Aged; D014442:Monophenol Monooxygenase / Q000097:blood / Q000235:genetics; D009363:Neoplasm Proteins / Q000097:blood / Q000235:genetics; D009367:Neoplasm Staging; D011446:Prospective Studies; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012878:Skin Neoplasms / Q000097:blood* / Q000473:pathology*; D058951:gp100 Melanoma Antigen
D000951:Antigens, Neoplasm; D015295:Antigens, Tumor-Associated, Carbohydrate; D014408:Biomarkers, Tumor; C072013:MAGEA3 protein, human; D058965:MART-1 Antigen; C074879:MIA antigen, human; C548667:MLANA protein, human; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C548637:PMEL protein, human; D012333:RNA, Messenger; D058951:gp100 Melanoma Antigen; D014442:Monophenol Monooxygenase