The detection of melanoma cells in peripheral blood by reverse transcription-polymerase chain reaction

PURPOSE Circulating melanoma cells can be detected in peripheral blood by means of tyrosinase mRNA amplification by reverse-transcriptase polymerase chain reaction (RT-PCR). We conducted a prospective study to evaluate the clinical significance of the presence of circulating neoplastic cells in the blood of patients with malignant melanoma (MM). METHODS A sensitive RT-PCR assay was used to detect tyrosinase mRNA in the peripheral blood of patients with stages I to IV melanoma. Healthy subjects or patients with other malignancies were used as negative controls. RESULTS Ninety-one assessable patients were included in the study. There was a statistically significant association between RT-PCR positivity and clinical stage. Circulating melanoma cells were detected in 36% of patients with localized disease (stages I and II), in 45% of patients with regional nodal involvement (stage III), and in 94% of patients with metastatic disease (stage IV) (P < .001). In stage II-III patients who were RT-PCR-positive for mRNA tyrosinase in blood, the recurrence rate and disease-free survival were significantly worse than patients who were RT-PCR-negative. In multivariate analysis, RT-PCR was an independent prognostic factor for recurrence in patients with nonmetastatic disease (P = .002). CONCLUSION The detection of circulating melanoma cells in peripheral blood by RT-PCR correlated with the clinical stage of patients with melanoma and was an independent prognostic factor for recurrence. Further studies are warranted to better assess the significance of this test in the evaluation of prognosis, early detection of relapse, and in monitoring the effectiveness of systemic therapy.

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Evaluation of the use of tyrosinase-specific and melanA/MART-1-specific reverse transcriptase-coupled-polymerase chain reaction to detect melanoma cells in peripheral blood samples from 299 patients with malignant melanoma

British Journal of Dermatology ◽

10.1046/j.1365-2133.2001.04015.x ◽

2001 ◽

Vol 144 (2) ◽

pp. 279-287 ◽

Cited By ~ 48

Author(s):

G.G. Brownbridge ◽

J. Gold ◽

M. Edward ◽

R.M. Mackie

Keyword(s):

Polymerase Chain Reaction ◽

Malignant Melanoma ◽

Reverse Transcriptase ◽

Peripheral Blood ◽

Melanoma Cells ◽

Chain Reaction ◽

Blood Samples ◽

Polymerase Chain

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Failure to Detect Measles Virus Rna, by Reverse Transcription-Polymerase Chain Reaction, in Peripheral Blood Leucocytes of Patients with Multiple Sclerosis

Multiple Sclerosis Journal ◽

10.1177/135245859600100403 ◽

1996 ◽

Vol 1 (4) ◽

pp. 204-206 ◽

Cited By ~ 7

Author(s):

B. Brankin ◽

M. Osman ◽

L. Herlihy ◽

S.A. Hawkins ◽

S.L. Cosby

Keyword(s):

Multiple Sclerosis ◽

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Peripheral Blood ◽

Measles Virus ◽

Chain Reaction ◽

Specific Primers ◽

Polymerase Chain ◽

Multiple Sclerosis Patients ◽

Peripheral Blood Leucocytes

We have examined peripheral blood leucocytes (PBLs) from 17 multiple sclerosis patients, two patients with rheumatoid arthritis, one case of acute childhood measles and one case of subacute sclerosing panencephalitis, as well as 19 healthy adult controls for measles virus (MV) RNA, by the technique of reverse transcription-polymerase chain reaction. MV nucleocapsid gene specific primers were used to amplify all PBL-derived cDNA samples. These proved to be negative with the exception of the sample derived from the acute measles case. Selected cases were examined further, using fusion gene and matrix gene specific primers. MV RNA could not be detected.

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Polymerase Chain Reaction-Based Detection of Circulating Melanoma Cells as an Effective Marker of Tumor Progression

Journal of Clinical Oncology ◽

10.1200/jco.1999.17.1.304 ◽

1999 ◽

Vol 17 (1) ◽

pp. 304-304 ◽

Cited By ~ 87

Author(s):

Giuseppe Palmieri ◽

Maria Strazzullo ◽

Paolo A. Ascierto ◽

Sabrina M.R. Satriano ◽

Antonio Daponte ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Tumor Progression ◽

Peripheral Blood ◽

Melanoma Cells ◽

Disease Stage ◽

Rt Pcr ◽

Chain Reaction ◽

Gene Markers ◽

Risk Of Recurrence ◽

Polymerase Chain

PURPOSE: 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. PATIENTS AND 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. CONCLUSION: 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.

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