Polymerase chain reaction-based detection of circulating melanoma cells as an effective marker of tumor progression. Melanoma Cooperative Group.
OBJECTIVE
Reverse transcriptase (RT) polymerase chain reaction (PCR) with multiple markers has been demonstrated to be highly sensitive in detecting circulating cells from patients with malignant melanoma (MM). We evaluated the clinical significance of the presence in peripheral blood of specific PCR-positive mRNA markers as an expression of circulating melanoma cells.


METHODS
Total cellular RNA was obtained from the peripheral blood of 235 patients with either localized (n = 154) or metastatic (n = 81) melanoma. We performed RT-PCR using tyrosinase, p97, MUC18, and MelanA/MART1 as gene markers. The PCR products were analyzed by gel electrophoresis and Southern blot hybridization. In addition, 20 healthy subjects and 21 patients with nonmelanoma cancer were used as negative controls.


RESULTS
Although detected at various levels among assessable patients, each mRNA marker was significantly correlated with disease stage. A significant correlation with disease stage was demonstrated for patients who were positive to all four markers (P < .0001) or to at least three markers (P < .001). Univariate analysis showed a significant correlation between risk of recurrence (evaluated in stage I, II, and III patients) and increasing number of PCR-positive markers (P = .0002). Logistic regression multivariate analysis indicated that each single marker (except tyrosinase) and, more especially, the presence of four PCR-positive markers remained statistically independent prognostic factors for tumor progression.


CONCLUSIONS
Our data establish the existence of a significant correlation among clinical stages, tumor progression, and presence of circulating melanoma-associated antigens in peripheral blood of MM patients. Preliminary assessment of a subset of patients with a higher risk of recurrence needs longer follow-up and further studies to define the role of RT-PCR in monitoring MM patients.
Palmieri
G
G
Department of Medicine, National Tumor Institute, Naples, Italy.
Strazzullo
M
M
Ascierto
P A
PA
Satriano
S M
SM
Daponte
A
A
Castello
G
G
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015703:Antigens, CD; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor / Q000097:blood; D051929:CD146 Antigen; D018450:Disease Progression; D005260:Female; D006801:Humans; D058965:MART-1 Antigen; D008297:Male; D008545:Melanoma / Q000276:immunology / Q000473:pathology* / Q000556:secondary; D058950:Melanoma-Specific Antigens; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics; D008875:Middle Aged; D014442:Monophenol Monooxygenase / Q000032:analysis / Q000235:genetics; D009363:Neoplasm Proteins / Q000032:analysis / Q000097:blood / Q000235:genetics; D009360:Neoplastic Cells, Circulating; D019006:Neural Cell Adhesion Molecules; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000097:blood; D012194:RNA-Directed DNA Polymerase; D012680:Sensitivity and Specificity; D012878:Skin Neoplasms / Q000276:immunology / Q000473:pathology*
D015703:Antigens, CD; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D051929:CD146 Antigen; D058965:MART-1 Antigen; C497902:MCAM protein, human; C548667:MLANA protein, human; D058950:Melanoma-Specific Antigens; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D019006:Neural Cell Adhesion Molecules; D012333:RNA, Messenger; D014442:Monophenol Monooxygenase; D012194:RNA-Directed DNA Polymerase