Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Annals of Surgical Oncology
Published in: Annals of Surgical Oncology 1/2009

01-01-2009 | Melanomas

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

Authors: Josep M. Hilari, Cristina Mangas, Liqiang Xi, Cristina Paradelo, Carlos Ferrándiz, Steven J. Hughes, Cindy Yueh, Ivy Altomare, William E. Gooding, Tony E. Godfrey

Published in: Annals of Surgical Oncology | Issue 1/2009

Login to get access

Abstract

The aim of this study was to evaluate the prognostic potential of quantitative reverse-transcription, polymerase chain reaction (qRT-PCR) in melanoma patients with pathologically negative sentinel lymph nodes (SLN). Our study included 195 node-negative melanoma patients with a Breslow thickness greater than 0.76 mm (n = 158), or less than 0.76 mm but who had Clark level IV–V, microscopic ulceration, or pathological signs of regression (n = 32), and five patients with melanoma of unknown thickness. SLNs were examined by serial-section histopathology. A portion of each SLN was frozen for qRT-PCR analysis using markers Tyrosinase, MART1, SSX2, MAGEA3, PAX3, and GalNAc-T. In addition, two other markers (PLAB and L1CAM) were evaluated for melanoma specificity but not for SLN analysis. Median follow-up was 64 months, during which time there were 15 (7.7%) recurrences. A total of 370 lymph nodes were analyzed by qRT-PCR. No association was found between quantitative expression level of any marker and disease recurrence. Previously published primer designs were tested for PAX3 and GalNAc-T and revealed that alternative PAX3 transcripts are differentially expressed in melanoma and benign lymph nodes. No associations with recurrence were found regardless of the transcripts amplified by different primer sets. PLAB and L1CAM did not appear to differentiate between malignant melanoma and benign melanocytes or lymph nodes in our analysis. We conclude that, in this large cohort of patients, multimarker qRT-PCR analysis of SLNs did not correlate with disease recurrence. Our data support specific PAX3 splice variants but not GalNAc-T, PLAB or L1CAM as possible markers for melanoma metastasis to SLNs.
Appendix
Available only for authorised users
Literature
1.
Morton DL, Wen DR, Wong JH, et al. Technical details of intraoperative lymphatic mapping for early stage melanoma. Arch Surg. 1992;127:392–9.CrossRefPubMed
2.
3.
Thompson JF, Uren RF. Lymphatic mapping in management of patients with primary cutaneous melanoma. Lancet Oncol. 2005;6:877–85.CrossRefPubMed
4.
Roberts AA, Cochran AJ. Pathologic analysis of sentinel lymph nodes in melanoma patients: current and future trends. J Surg Oncol. 2004;85:152–61.CrossRefPubMed
5.
Spanknebel K, Coit DG, Bieligk SC, et al. Characterization of micrometastatic disease in melanoma sentinel lymph nodes by enhanced pathology: recommendations for standardizing pathologic analysis. Am J Surg Pathol. 2005;29:305–17.CrossRefPubMed
6.
Yee VS, Thompson JF, McKinnon JG, et al. Outcome in 846 cutaneous melanoma patients from a single center after a negative sentinel node biopsy. Ann Surg Oncol. 2005;12:429–39.CrossRefPubMed
7.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
8.
Kammula US, Ghossein R, Bhattacharya S, Coit DG. Serial follow-up and the prognostic significance of reverse transcriptase-polymerase chain reaction-staged sentinel lymph nodes from melanoma patients. J Clin Oncol. 2004;22:3989–96.CrossRefPubMed
9.
Shivers SC, Wang X, Li W, et al. Molecular staging of malignant melanoma: correlation with clinical outcome. JAMA. 1998;280:1410–5.CrossRefPubMed
10.
Blaheta HJ, Ellwanger U, Schittek B, et al. Examination of regional lymph nodes by sentinel node biopsy and molecular analysis provides new staging facilities in primary cutaneous melanoma. J Invest Dermatol. 2000;114:637–42.CrossRefPubMed
11.
Kammula US, Ghossein R, Bhattacharya S, Coit DG. Serial follow-up and the prognostic significance of reverse transcriptase-polymerase chain reaction-staged sentinel lymph nodes from melanoma patients. J Clin Oncol. 2004;22:3989–96.CrossRefPubMed
12.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
13.
Scoggins CR, Ross MI, Reintgen DS, et al. Prospective multi-institutional study of reverse transcriptase polymerase chain reaction for molecular staging of melanoma. J Clin Oncol. 2006;24:2849–57.CrossRefPubMed
14.
Goydos JS, Patel KN, Shih WJ, et al. Patterns of recurrence in patients with melanoma and histologically negative but RT-PCR-positive sentinel lymph nodes. J Am Coll Surg. 2003;196:196–204.CrossRefPubMed
15.
Ribuffo D, Gradilone A, Vonella M, et al. Prognostic significance of reverse transcriptase-polymerase chain reaction-negative sentinel nodes in malignant melanoma. Ann Surg Oncol. 2003;10:396–402.CrossRefPubMed
16.
Ulrich J, Bonnekoh B, Bockelmann R, et al. Prognostic significance of detecting micrometastases by tyrosinase RT/PCR in sentinel lymph node biopsies: lessons from 322 consecutive melanoma patients. Eur J Cancer. 2004;40:2812–9.CrossRefPubMed
17.
Mangas C, Hilari JM, Paradelo C, et al. Prognostic significance of molecular staging study of sentinel lymph nodes by reverse transcriptase-polymerase chain reaction for tyrosinase in melanoma patients. Ann Surg Oncol. 2006;13:910–8.CrossRefPubMed
18.
Gradilone A, Ribuffo D, Silvestri I, et al. Detection of melanoma cells in sentinel lymph nodes by reverse transcriptase-polymerase chain reaction: prognostic significance. Ann Surg Oncol. 2004;11:983–7.CrossRefPubMed
19.
Mocellin S, Hoon DS, Pilati P, Rossi CR, Nitti D. Sentinel lymph node molecular ultrastaging in patients with melanoma: a systematic review and meta-analysis of prognosis. J Clin Oncol. 2007;25:1588–95.CrossRefPubMed
20.
Scoggins CR, Ross MI, Reintgen DS, et al. Prospective multi-institutional study of reverse transcriptase polymerase chain reaction for molecular staging of melanoma. J Clin Oncol. 2006;24:2849–57.CrossRefPubMed
21.
Abrahamsen HN, Sorensen BS, Nexo E, et al. Pathologic assessment of melanoma sentinel nodes: a role for molecular analysis using quantitative real-time reverse transcription-PCR for MART–1 and tyrosinase messenger RNA. Clin Cancer Res. 2005;11:1425–33.CrossRefPubMed
22.
Tucker MA, Fraser MC, Goldstein AM, et al. A natural history of melanomas and dysplastic nevi: an atlas of lesions in melanoma-prone families. Cancer. 2002;94:3192–209.CrossRefPubMed
23.
Holt JB, Sangueza OP, Levine EA, et al. Nodal melanocytic nevi in sentinel lymph nodes. Correlation with melanoma-associated cutaneous nevi. Am J Clin Pathol. 2004;121:58–63.CrossRefPubMed
24.
Starz H, Haas CJ, Schulz GM, Balda BR. Tyrosinase RT-PCR as a supplement to histology for detecting melanoma and nevus cells in paraffin sections of sentinel lymph nodes. Mod Pathol. 2003;16:920–9.CrossRefPubMed
25.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
26.
Talantov D, Mazumder A, Yu JX, et al. Novel genes associated with malignant melanoma but not benign melanocytic lesions. Clin Cancer Res. 2005;11:7234–42.CrossRefPubMed
27.
Rex J, Paradelo C, Mangas C, et al. Single-institution experience in the management of patients with clinical stage I and II cutaneous melanoma: results of sentinel lymph node biopsy in 240 cases. Dermatol Surg. 2005;31:1385–93.PubMed
28.
Kirkwood JM, Ibrahim JG, Sondak VK, et al. High- and low-dose interferon alfa-2b in high-risk melanoma: first analysis of intergroup trial E1690/S9111/C9190. J Clin Oncol. 2000;18:2444–58.PubMed
29.
Tassone F, Hagerman RJ, Taylor AK, et al. Elevated levels of FMR1 mRNA in carrier males: a new mechanism of involvement in the fragile-X syndrome. Am J Hum Genet. 2000;66:6–15.CrossRefPubMedPubMedCentral
30.
Talantov D, Mazumder A, Yu JX, et al. Novel genes associated with malignant melanoma but not benign melanocytic lesions. Clin Cancer Res. 2005;11:7234–42.CrossRefPubMed
31.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
32.
Yee VS, Thompson JF, McKinnon JG, et al. Outcome in 846 cutaneous melanoma patients from a single center after a negative sentinel node biopsy. Ann Surg Oncol. 2005;12:429–39.CrossRefPubMed
33.
Kammula US, Ghossein R, Bhattacharya S, Coit DG. Serial follow-up and the prognostic significance of reverse transcriptase-polymerase chain reaction-staged sentinel lymph nodes from melanoma patients. J Clin Oncol. 2004;22:3989–96.CrossRefPubMed
34.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
35.
Li W, Stall A, Shivers SC, et al. Clinical relevance of molecular staging for melanoma: comparison of RT-PCR and immunohistochemistry staining in sentinel lymph nodes of patients with melanoma. Ann Surg. 2000;231:795–803.CrossRefPubMedPubMedCentral
36.
Shivers SC, Wang X, Li W, et al. Molecular staging of malignant melanoma: correlation with clinical outcome. JAMA. 1998;280:1410–5.CrossRefPubMed
37.
Blaheta HJ, Ellwanger U, Schittek B, et al. Examination of regional lymph nodes by sentinel node biopsy and molecular analysis provides new staging facilities in primary cutaneous melanoma. J Invest Dermatol. 2000;114:637–42.CrossRefPubMed
38.
Ulrich J, Bonnekoh B, Bockelmann R, et al. Prognostic significance of detecting micrometastases by tyrosinase RT/PCR in sentinel lymph node biopsies: lessons from 322 consecutive melanoma patients. Eur J Cancer. 2004;40:2812–9.CrossRefPubMed
39.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
40.
Kuo CT, Hoon DS, Takeuchi H, et al. Prediction of disease outcome in melanoma patients by molecular analysis of paraffin-embedded sentinel lymph nodes. J Clin Oncol. 2003;21:3566–72.CrossRefPubMed
41.
Goydos JS, Patel KN, Shih WJ, et al. Patterns of recurrence in patients with melanoma and histologically negative but RT-PCR-positive sentinel lymph nodes. J Am Coll Surg. 2003;196:196–204.CrossRefPubMed
42.
Ribuffo D, Gradilone A, Vonella M, et al. Prognostic significance of reverse transcriptase-polymerase chain reaction-negative sentinel nodes in malignant melanoma. Ann Surg Oncol. 2003;10:396–402.CrossRefPubMed
43.
Bostick PJ, Morton DL, Turner RR, et al. Prognostic significance of occult metastases detected by sentinel lymphadenectomy and reverse transcriptase-polymerase chain reaction in early-stage melanoma patients. J Clin Oncol. 1999;17:3238–44.PubMed
44.
Rimoldi D, Lemoine R, Kurt AM, et al. Detection of micrometastases in sentinel lymph nodes from melanoma patients: direct comparison of multimarker molecular and immunopathological methods. Melanoma Res. 2003;13:511–20.CrossRefPubMed
45.
Giese T, Engstner M, Mansmann U, Hartschuh W, Arden B. Quantification of melanoma micrometastases in sentinel lymph nodes using real-time RT-PCR. J Invest Dermatol. 2005;124:633–7.CrossRefPubMed
46.
Mangas C, Hilari JM, Paradelo C, et al. Prognostic significance of molecular staging study of sentinel lymph nodes by reverse transcriptase-polymerase chain reaction for tyrosinase in melanoma patients. Ann Surg Oncol. 2006;13:910–8.CrossRefPubMed
47.
Kammula US, Ghossein R, Bhattacharya S, Coit DG. Serial follow-up and the prognostic significance of reverse transcriptase-polymerase chain reaction–staged sentinel lymph nodes from melanoma patients. J Clin Oncol. 2004;22:3989–96.CrossRefPubMed
48.
Scoggins CR, Ross MI, Reintgen DS, et al. Prospective multi-institutional study of reverse transcriptase polymerase chain reaction for molecular staging of melanoma. J Clin Oncol. 2006;24:2849–57.CrossRefPubMed
49.
Mocellin S, Hoon DS, Pilati P, Rossi CR, Nitti D. Sentinel lymph node molecular ultrastaging in patients with melanoma: a systematic review and meta-analysis of prognosis. J Clin Oncol. 2007;25:1588–95.CrossRefPubMed
50.
Scoggins CR, Ross MI, Reintgen DS, et al. Prospective multi-institutional study of reverse transcriptase polymerase chain reaction for molecular staging of melanoma. J Clin Oncol. 2006;24:2849–57.CrossRefPubMed
51.
Kammula US, Ghossein R, Bhattacharya S, Coit DG. Serial follow-up and the prognostic significance of reverse transcriptase-polymerase chain reaction–staged sentinel lymph nodes from melanoma patients. J Clin Oncol. 2004;22:3989–96.CrossRefPubMed
52.
Mangas C, Hilari JM, Paradelo C, et al. Prognostic significance of molecular staging study of sentinel lymph nodes by reverse transcriptase-polymerase chain reaction for tyrosinase in melanoma patients. Ann Surg Oncol. 2006;13:910–8.CrossRefPubMed
53.
Li W, Stall A, Shivers SC, et al. Clinical relevance of molecular staging for melanoma: comparison of RT-PCR and immunohistochemistry staining in sentinel lymph nodes of patients with melanoma. Ann Surg. 2000;231:795–803.CrossRefPubMedPubMedCentral
54.
Blaheta HJ, Ellwanger U, Schittek B, et al. Examination of regional lymph nodes by sentinel node biopsy and molecular analysis provides new staging facilities in primary cutaneous melanoma. J Invest Dermatol. 2000;114:637–42.CrossRefPubMed
55.
Goydos JS, Patel KN, Shih WJ, et al. Patterns of recurrence in patients with melanoma and histologically negative but RT-PCR-positive sentinel lymph nodes. J Am Coll Surg. 2003;196:196–204.CrossRefPubMed
56.
Ribuffo D, Gradilone A, Vonella M, et al. Prognostic significance of reverse transcriptase-polymerase chain reaction-negative sentinel nodes in malignant melanoma. Ann Surg Oncol. 2003;10:396–402.CrossRefPubMed
57.
Ulrich J, Bonnekoh B, Bockelmann R, et al. Prognostic significance of detecting micrometastases by tyrosinase RT/PCR in sentinel lymph node biopsies: lessons from 322 consecutive melanoma patients. Eur J Cancer. 2004;40:2812–9.CrossRefPubMed
58.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
59.
Giese T, Engstner M, Mansmann U, Hartschuh W, Arden B. Quantification of melanoma micrometastases in sentinel lymph nodes using real-time RT-PCR. J Invest Dermatol. 2005;124:633–7.CrossRefPubMed
60.
Takeuchi H, Morton DL, Kuo C, et al. Prognostic significance of molecular upstaging of paraffin-embedded sentinel lymph nodes in melanoma patients. J Clin Oncol. 2004;22:2671–80.CrossRefPubMedPubMedCentral
61.
Giese T, Engstner M, Mansmann U, Hartschuh W, Arden B. Quantification of melanoma micrometastases in sentinel lymph nodes using real-time RT-PCR. J Invest Dermatol. 2005;124:633–7.CrossRefPubMed
62.
Balch CM, Soong SJ, Atkins MB, et al. An evidence-based staging system for cutaneous melanoma. CA Cancer J Clin. 2004;54:131–49.CrossRefPubMed
63.
Talantov D, Mazumder A, Yu JX, et al. Novel genes associated with malignant melanoma but not benign melanocytic lesions. Clin Cancer Res. 2005;11:7234–42.CrossRefPubMed
Metadata
Title
Molecular Staging of Pathologically Negative Sentinel Lymph Nodes from Melanoma Patients Using Multimarker, Quantitative Real-Time RT-PCR
Authors
Josep M. Hilari
Cristina Mangas
Liqiang Xi
Cristina Paradelo
Carlos Ferrándiz
Steven J. Hughes
Cindy Yueh
Ivy Altomare
William E. Gooding
Tony E. Godfrey
Publication date
01-01-2009
Publisher
Springer New York
Published in
Annals of Surgical Oncology / Issue 1/2009
Print ISSN: 1068-9265
Electronic ISSN: 1534-4681
DOI
https://doi.org/10.1245/s10434-008-0183-9

Other articles of this Issue 1/2009

Annals of Surgical Oncology 1/2009 Go to the issue

Breast Oncology

International Randomized Trials’ Attention to Locoregional Breast Cancer Control and Promise of Personal Genomics and Whole-Genome Scans for Personalized Approaches

Breast Oncology

Impact and Outcomes of Routine Microstaging of Sentinel Lymph Nodes in Breast Cancer: Significance of the pN0(i+) and pN1mi Categories

Gynecologic Oncology

Matched-Case Comparison for the Role of Surgery in FIGO Stage Ib1–IIa Squamous Cell Carcinoma of Cervix and Suspicious Para-Aortic Lymph Node Metastasis

Hepatobiliary and Pancreatic Tumors

A Proposed Staging System for Intrahepatic Cholangiocarcinoma

Breast Oncology

Is Electron Beam Intraoperative Radiotherapy (ELIOT) Safe in Pregnant Women with Early Breast Cancer? In Vivo Dosimetry to Assess Fetal Dose

Gastrointestinal Oncology

Perineal Healing and Survival After Anal Cancer Salvage Surgery: 10-Year Experience with Primary Perineal Reconstruction Using the Vertical Rectus Abdominis Myocutaneous (VRAM) Flap