Facts and Pitfalls in the Detection of Tyrosinase mRNA in the Blood of Melanoma Patients by RT-PCR

2001 ◽  
pp. 105-112 ◽  
Author(s):  
S. Seiter ◽  
G. Rappl ◽  
W. Tilgen ◽  
S. Ugurel ◽  
U. Reinhold
Keyword(s):  
Rt Pcr ◽  
Melanoma Patients

Detection of Micrometastases in Sentinel Lymph Nodes of Melanoma Patients by RT-PCR.

2002 ◽  
Vol 64 (2) ◽  
pp. 211-217 ◽  
Author(s):  
Yoichi MOROI ◽  
Chikage OBATA ◽  
Shohei FUJITA ◽  
Satoko SHIBATA ◽  
Sawako KONISHI ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Sentinel Lymph Nodes ◽  
Rt Pcr ◽  
Melanoma Patients

Molecular Staging in Stage II and III Melanoma Patients and Its Effect on Long-Term Survival

2005 ◽  
Vol 23 (6) ◽  
pp. 1218-1227 ◽  
Author(s):  
Christiane Voit ◽  
Martina Kron ◽  
Juergen Rademaker ◽  
Markus Schwürzer-Voit ◽  
Wolfram Sterry ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Prognostic Value ◽  
Stage Ii ◽  
Disease Specific Survival ◽  
Rt Pcr ◽  
Term Survival ◽  
Histologic Subtype ◽  
Melanoma Patients ◽  
Disease Specific

Purpose To assess the prognostic value of serial reverse transcriptase polymerase chain reaction (RT-PCR) -based measurements of tyrosinase mRNA in peripheral blood of stage II and III melanoma patients. Patients and Methods During routine follow-up of American Joint Committee on Cancer stage II and III melanoma patients, serial testing for tyrosinase transcripts in peripheral blood was performed by RT-PCR. The PCR results were compared with the clinical data collected during the follow-up. Results Over a period of 3 years, 111 patients (78 stage II and 33 stage III patients) were enrolled, and tyrosinase determinations were carried out. The 6-year disease-specific survival probability was 97% for patients always showing negative RT-PCR results and 67% for patients who tested positive at least once. In a Cox proportional hazards model, the prognostic value of sex, age, site of primary tumor, histologic subtype, stage, Breslow's tumor thickness, Clark level, and the time-dependent variable PCR result was assessed. Patients with a positive RT-PCR test had a distinctly higher risk of dying from melanoma, with a hazard ratio of 12.6 (95% CI, 3.4 to 46.3; P < .001). Conclusion Our study shows a strong association between PCR and disease-specific survival time. Detection of tyrosinase mRNA in peripheral blood may be of similar importance for the clinical course of melanoma as the detection of micrometastatic disease in the sentinel lymph node. Whether a combination of these two factors leads to a better definition of the prognosis of melanoma patients is under investigation in current studies.

Prognostic value of tyrosinase RT-PCR analysis in histologically negative sentinel lymph nodes from melanoma patients

2004 ◽  
Vol 22 (14_suppl) ◽  
pp. 7562-7562
Author(s):  
M. Gonzalez Cao ◽  
C. Badenas ◽  
J. Malvehy ◽  
R. Martí ◽  
T. Castel ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Prognostic Value ◽  
Sentinel Lymph Nodes ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Melanoma Patients

Molecular Staging of Pathologically Negative Sentinel Lymph Nodes from Melanoma Patients Using Multimarker, Quantitative Real-Time RT-PCR

2009 ◽  
Vol 16 (1) ◽  
Author(s):  
Josep M. Hilari ◽  
Cristina Mangas ◽  
Liqiang Xi ◽  
Cristina Paradelo ◽  
Carlos Ferrándiz ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
Real Time ◽  
Sentinel Lymph Nodes ◽  
Rt Pcr ◽  
Molecular Staging ◽  
Melanoma Patients

Tyrosinase mRNA in Blood of Patients With Melanoma Treated With Adjuvant Interferon

2002 ◽  
Vol 20 (19) ◽  
pp. 4032-4039 ◽  
Author(s):  
Begoña Mellado ◽  
Maria del Carmen Vela ◽  
Dolors Colomer ◽  
Lorena Gutierrez ◽  
Teresa Castel ◽  
...  
Keyword(s):  
Interferon Therapy ◽  
Surrogate Marker ◽  
Disease Free Survival ◽  
Melanoma Cells ◽  
Interferon Response ◽  
Interferon Treatment ◽  
Rt Pcr ◽  
Melanoma Patients ◽  
Adjuvant Interferon

PURPOSE: To evaluate the clinical significance of the detection of circulating melanoma cells in patients treated with adjuvant interferon and to determine their potential value as a marker of interferon response. PATIENTS AND METHODS: We prospectively analyzed 616 peripheral-blood samples from 120 melanoma patients with stage IIA (n = 33), IIB (n = 22), III (n = 50), or IV (surgically resected) (n = 15) disease receiving adjuvant interferon alfa-2b therapy. Tyrosinase mRNA was assayed by reverse transcriptase polymerase chain reaction (RT-PCR) as a marker of circulating melanoma cells before the start of interferon and every 2 to 3 months thereafter. RESULTS: With a median follow-up time of 32.3 months (range, 7.1 to 77.5 months), 47 patients (39.8%) relapsed and 31 (26%) died. During adjuvant interferon treatment, 76 patients (64%) had undetected circulating melanoma cells and 44 patients (36%) had a positive RT-PCR result in at least one sample. Actuarial 5-year disease-free survival was 62% in patients with persistently negative RT-PCR during interferon treatment and 38% for patients with positive RT-PCR during interferon (P = .02). Actuarial 5-year overall survival was 75% and 50%, respectively (P = .03). CONCLUSION: Patients with melanoma and tyrosinase mRNA detected in the blood during adjuvant interferon therapy had a worse prognosis compared with patients with undetected tyrosinase mRNA during treatment. Further investigation into the detection of circulating melanoma cells as a surrogate marker of response to adjuvant interferon therapy is warranted.

scholarly journals The Analysis of Tyrosinase-Specific mRNA in Blood Samples of Melanoma Patients by RT-PCR Is Not a Useful Test for Metastic Tumor Progression

1997 ◽  
Vol 108 (2) ◽  
pp. 166-169 ◽  
Author(s):  
Uwe Reinhold ◽  
Hans-Christian Lüdtke-Handjery ◽  
Sylvia Schnautz ◽  
Hans-Wilhelm Kreysel ◽  
Hinrich Abken
Keyword(s):  
Tumor Progression ◽  
Rt Pcr ◽  
Blood Samples ◽  
Melanoma Patients ◽  
Specific Mrna

Prognostic Significance of Occult Metastases Detected by Sentinel Lymphadenectomy and Reverse Transcriptase–Polymerase Chain Reaction in Early-Stage Melanoma Patients

1999 ◽  
Vol 17 (10) ◽  
pp. 3238-3244 ◽  
Author(s):  
Peter J. Bostick ◽  
Donald L. Morton ◽  
Roderick R. Turner ◽  
Kelly T. Huynh ◽  
He-Jing Wang ◽  
...  
Keyword(s):  
Early Stage ◽  
Disease Relapse ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Sentinel Lymphadenectomy ◽  
Increased Risk ◽  
Melanoma Metastases ◽  
Melanoma Patients ◽  
Polymerase Chain ◽  
Mrna Markers

PURPOSE: Detection of micrometastases in the regional tumor-draining lymph nodes is critical for accurate staging and prognosis in melanoma patients. We hypothesized that a multiple-mRNA marker (MM) reverse transcriptase–polymerase chain reaction (RT-PCR) assay would improve the detection of occult metastases in the sentinel node (SN), compared with hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC), and that MM expression is predictive of disease relapse. PATIENTS AND METHODS: Seventy-two consecutive patients with clinical early-stage melanoma underwent sentinel lymphadenectomy (SLND). Their SNs were serially sectioned and assessed for MAGE-3, MART-1, and tyrosinase mRNA expression by RT-PCR, in parallel with H&E staining and IHC, for melanoma metastases. MM expression in the SNs was correlated with H&E and IHC assay results, standard prognostic factors, and disease-free survival. RESULTS: In 17 patients with H&E- and/or IHC-positive SNs, 16 (94%) expressed two or more mRNAmarkers. Twenty (36%) of 55 patients with histopathologically negative SNs expressed two or more mRNA markers. By multivariate analysis, patients at increased risk of metastases to the SN had thicker lesions (P = .03), were 60 years of age or younger (P < .05), and/or were MM-positive (P < .001). Patients with histopathologically melanoma-free SNs who were MM-positive, compared with those who were positive for one or fewer mRNA markers, were at increased risk of recurrence (P = .02). Patients who were MM-positive with histopathologically proven metastases in the SN were at greatest risk of disease relapse (P = .01). CONCLUSION: H&E staining and IHC underestimate the true incidence of melanoma metastases. MM expression in the SN more accurately reflects melanoma micrometastases and is also a more powerful predictor of disease relapse than are H&E staining and IHC alone.

Survivin, bcl-2, bax, and bcl-X Gene Expression in Sentinel Lymph Nodes From Melanoma Patients

2003 ◽  
Vol 21 (2) ◽  
pp. 306-312 ◽  
Author(s):  
Angela Gradilone ◽  
Paola Gazzaniga ◽  
Diego Ribuffo ◽  
Susanna Scarpa ◽  
Emanuele Cigna ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lymph Nodes ◽  
Survivin Expression ◽  
Sentinel Lymph Nodes ◽  
Rt Pcr ◽  
Survivin Gene ◽  
Variable Expression ◽  
Melanoma Patients ◽  
X Gene ◽  
Disease Free

Purpose: The expression of apoptosis-related genes, such as survivin, bcl-2, bcl-X, and bax, has been evaluated by reverse transcriptase polymerase chain reaction (RT-PCR) and by immunohistochemistry in sentinel lymph nodes (SLNs) from melanoma patients and then correlated to the outcome of patients. Patients and Methods: Thirty-six SLNs were examined. After RNA extraction, an RT-PCR followed by Southern blot hybridization was performed to detect survivin, bcl-2, bcl-X, and bax mRNA. bcl-2, survivin, and bax gene expression was evaluated, whenever possible, also by immunohistochemistry at the protein level. Results: We found a significant correlation (P < .005) between survivin expression and outcome of patients; in fact, 61.5% of patients expressing survivin gene progressed or died because of the disease, whereas 38.5% are currently disease-free. Among patients negative for survivin expression, 100% are disease-free after a median follow-up time of 52.9 months. We did not find a significant correlation between bcl-2, bax, and bcl-X gene expression and outcome of patients. In fact, these genes were found equally expressed in patients with disease progression and in disease-free patients. Conclusion: Our findings show a variable expression of apoptosis-related genes in SLNs of melanoma patients; more interestingly, we found that survivin expression correlates to outcome of patients in a statistically significant way, whereas the expression of other genes, such as bcl-2, bax, and bcl-X, did not seem to correlate to progression of disease. We suggest that the detection of survivin gene expression by RT-PCR in SLNs may be a useful prognostic indicator.

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