Annals of Surgical Oncology

, Volume 6, Issue 3, pp 232–240 | Cite as

Detection of Tyrosinase mRNA by Reverse Transcription Polymerase Chain Reaction in Melanoma Sentinel Nodes

  • Samuel C. Bieligk
  • Ronald Ghossein
  • Satyajit Bhattacharya
  • Daniel G. Coit
Original Article


Background: Sentinel lymph node (SLN) biopsy is an alternative to elective dissection or observation for management of lymph node basins in patients with cutaneous melanomas. The detection of tyrosinase mRNA in melanoma SLN specimens by reverse transcription-polymerase chain reaction (RT-PCR) has been reported to be a more sensitive method to detect subclinical metastases, compared with histological analysis. The aims of this study were to (1) define the yield of RT-PCR in assessing SLNs, compared with histological analysis, (2) identify the incidence of false-positive results in SLNs, and (3) report the rate of actin PCR negativity (i.e., samples with degraded RNA) in SLNs.

Methods: Twenty-eight patients with 1.2–9.6-mm cutaneous melanomas underwent SLN biopsy (between October 1996 and March 1997). One half of each SLN was analyzed by nested RT-PCR for tyrosinase mRNA. The other half of the SLN was examined by routine microscopy. Twenty-one lymph nodes from patients without melanoma were evaluated as controls.

Results: Two of the 28 patients with melanoma were excluded because of RNA degradation, as indicated by actin negativity. Six of the remaining 26 patients exhibited melanoma metastases in routine histological examinations. All histologically positive lymph nodes were RT-PCR-positive. Thirteen of the 20 (65%) histologically negative cases were RT-PCR-positive. Of 21 control lymph nodes, 3 were actin-negative and were not assessable for tyrosinase mRNA. Two of the remaining 18 (11%) negative-control nodes were RT-PCR-positive.

Conclusions: Among patients undergoing SLN biopsy, tyrosinase mRNA was detectable in 73% of SLNs from patients at risk for regional nodal metastases, including all of those with histologically positive SLNs. There is a definable false-positive rate for tyrosinase mRNA detection in the lymph nodes of patients who do not have melanoma. Actin verification of RNA integrity is necessary to ensure the accuracy of this test in detecting tyrosinase mRNA. Ongoing follow-up monitoring will define the prognostic value of this assay.

Key Words

Tyrosinase mRNA Reverse transcriptase polymerase chain reaction Sentinel lymph node 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Sim FH, Taylor WF, Pritchard DJ, Soule EH. Lymphadenectomy in the management of stage I malignant melanoma: a prospective randomized study. Mayo Clin Proc 1986;61:697–705.PubMedGoogle Scholar
  2. 2.
    Veronesi U, Adamus J, Bandiera DC, et al. Inefficacy of immediate node dissection in stage I melanoma of the limbs. N Engl J Med 1977;297:627–630.PubMedCrossRefGoogle Scholar
  3. 3.
    Veronesi U, Adamus J, Bandiera DC, et al. Delayed regional lymph node dissection in stage I melanoma of the skin of the lower extremities. Cancer 1982;49:2420–2430.PubMedGoogle Scholar
  4. 4.
    Balch CM, Soong SJ, Bartolucci AA, et al. Efficacy of an elective regional lymph node dissection of 1 to 4 mm thick melanomas for patients 60 years of age and younger. Ann Surg 1996;224:255–266.PubMedGoogle Scholar
  5. 5.
    Dickler MN, Coit DG, Meyers ML. Adjuvant therapy of malignant melanoma. Surg Oncol Clin North Am 1997;6:793–812.Google Scholar
  6. 6.
    Morton DL, Wen DR, Wong JH, et al. Technical details of intraoperative lymphatic mapping for early stage melanoma. Arch Surg 1992;127:392–399.PubMedGoogle Scholar
  7. 7.
    Multicenter Selective Lymphadenectomy Trial. National Cancer Institute Grant PO1-CA29605–12.Google Scholar
  8. 8.
    Brady MS, Coit DG. Lymphatic mapping in the management of the patient with cutaneous melanoma. The Cancer Journal 1997;10:87–93.Google Scholar
  9. 9.
    Cochran AJ, Wen DR, Morton DL. Occult tumor cells in the lymph nodes of patients with pathological stage I malignant melanoma. Am J Surg Pathol 1988;12:612–618.CrossRefPubMedGoogle Scholar
  10. 10.
    Bieligk SC, Klimstra DS, Rosai J, Coit DG. The detection of micrometastases in melanoma sentinel nodes by step sectioning and immunohistochemical staining. Melanoma Res 1997;7(Suppl 1):S125 [Abstract].Google Scholar
  11. 11.
    Ghossein RA, Coit DG, Brennan MF, et al. Prognostic significance of peripheral blood and bone marrow tyrosinase messenger RNA in malignant melanoma. Clin Cancer Res 1998;4:419–428.PubMedGoogle Scholar
  12. 12.
    Hoon DSB, Wang Y, Dale PS, et al. Detection of occult melanoma cells in blood with a multiple-marker polymerase chain reaction assay. J Clin Oncol 1995;13:2109–2116.PubMedGoogle Scholar
  13. 13.
    Moreno JG, Croce CM, Fisher R, et al. Detection of hematogenous micrometastases in patients with prostate cancer. Cancer Res 1992;52:6110–6112.PubMedGoogle Scholar
  14. 14.
    Smith B, Selby P, Southgate J, Pittman K, Bradley C, Blair GE. Detection of melanoma cells in peripheral blood by means of reverse transcriptase and polymerase chain reaction. Lancet 1991;338:1227–1229.PubMedGoogle Scholar
  15. 15.
    Kawasaki ES, Clark SS, Coyne MY, et al. Diagnosis of chronic myeloid and acute lymphocytic leukemias by detection of leukemia- specific mRNA sequences amplified in vitro. Proc Natl Acad Sci USA 1988;85:5698–5702.PubMedGoogle Scholar
  16. 16.
    Tada M, Omata M, Kawai S, et al. Detection of ras gene mutations in pancreatic juice and peripheral blood of patients with pancreatic adenocarcinoma. Cancer Res 1993;53:2472–2474.PubMedGoogle Scholar
  17. 17.
    Wang X, Heller R, Van Voorhis N, et al. Detection of submicroscopic lymph node metastases with polymerase chain reaction in patients with malignant melanoma. Ann Surg 1994;220:768–774.PubMedGoogle Scholar
  18. 18.
    Reintgen D, Balch CM, Kirkwood J, Ross MI. Recent advances in the care of the patient with malignant melanoma. Ann Surg 1997;225:1–14.PubMedGoogle Scholar
  19. 19.
    Van der Velde-Zimmermann D, Roijers JFM, Bouwens-Rombouts A, et al. Molecular test for the detection of tumor cells in blood and sentinel nodes of melanoma patients. Am J Pathol 1996;149:759–764.PubMedGoogle Scholar
  20. 20.
    Battyani Z, Xerri L, Hassoun J, Bonerandi J-J, Grob J-J. Tyrosinase gene expression in human tissues. Pigment Cell Res 1993;6:400–408.PubMedGoogle Scholar
  21. 21.
    Naber SP, Smith LL, Wolfe HJ. Role of the frozen tissue bank in molecular pathology. Diagn Mol Pathol 1992;1:73–79.PubMedGoogle Scholar
  22. 22.
    Ben-Ezra J, Johnson DA, Rossi J, Cook N, Wu A. Effect of fixation on the amplification of nucleic acids from paraffin embedded material by the polymerase chain reaction. J Histochem Cytochem 1991;39:351–354.PubMedGoogle Scholar
  23. 23.
    Fleming ID, Cooper JS, Henson DE, et al., eds. American Joint Committee on Cancer cancer staging manual, 5th edn. Philadelphia: JB Lippincott, 1997.Google Scholar
  24. 24.
    McMasters KM, Reintgen DS, Ross MI, Edwards MJ, Cerrito P. Sunbelt Melanoma Trial: A multicenter trial of adjuvant interferon alfa-2b for melanoma patients with early lymph node metastasis detected by lymphatic mapping and sentinel lymph node biopsy. Louisville, KY: 1996.Google Scholar
  25. 25.
    Mattano LA, Moss TJ, Emerson SG. Sensitive detection of rare circulating neuroblastoma cells by the reverse transcriptase-polymerase chain reaction. Cancer Res 1992;52:4701–4705.PubMedGoogle Scholar
  26. 26.
    Kaplan JC, Kahn A, Chelly J. Illegitimate transcription: its use in the study of inherited disease. Hum Mutat 1992;1:357–360.PubMedGoogle Scholar
  27. 27.
    Mellado B, Colomer D, Castel T, et al. Detection of circulating neoplastic cells by reverse-transcriptase polymerase chain reaction in malignant melanoma: association with clinical stage and prognosis. J Clin Oncol 1996;14:2091–2097.PubMedGoogle Scholar
  28. 28.
    Bautista NC, Cohen S, Anders KH. Benign melanocytic nevus cells in axillary lymph nodes: a prospective incidence and immunohistochemical study with literature review. Am J Clin Pathol 1994;102:102–108.PubMedGoogle Scholar
  29. 29.
    Carson KF, Wen D-R, Li P-X, et al. Nodal nevi and cutaneous melanomas. Am J Surg Pathol 1996;20:834–840.PubMedGoogle Scholar
  30. 30.
    Buzaid AC, Balch CM. Polymerase chain reaction for detection of melanoma in peripheral blood: too early to assess clinical value. J Natl Cancer Inst 1996;88:569–570.PubMedGoogle Scholar
  31. 31.
    Reintgen D, Albertini J, Berman C, et al. Accurate nodal staging of malignant melanoma. Cancer Control, J Moffitt Cancer Center 1995;2:405–414.Google Scholar

Copyright information

© The Society of Surgical Oncology, Inc. 1999

Authors and Affiliations

  • Samuel C. Bieligk
    • 1
  • Ronald Ghossein
    • 2
  • Satyajit Bhattacharya
    • 2
  • Daniel G. Coit
    • 3
  1. 1.Department of Surgery, Division of Surgical OncologyUniversity of Texas Southwestern Medical Center at DallasDallas
  2. 2.Department of PathologyMemorial Sloan-Kettering Cancer CenterNew York
  3. 3.Gastric and Mixed Tumor ServiceMemorial Sloan-Kettering Cancer CenterNew York

Personalised recommendations