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. 1996 Sep;149(3):759–764.

Molecular test for the detection of tumor cells in blood and sentinel nodes of melanoma patients.

D Van der Velde-Zimmermann 1, J F Roijers 1, A Bouwens-Rombouts 1, R A De Weger 1, P W De Graaf 1, M G Tilanus 1, J G Van den Tweel 1
PMCID: PMC1865135  PMID: 8780380

Abstract

Lymph node metastasis dramatically decreases the 5-year survival of melanoma patients. The so-called sentinel node surgery offers a therapeutic approach to resect the first draining lymph node. This technique enables accurate staging of melanoma patients in an early stage of the disease. Detection of a sentinel node metastasis is a strong argument for local lymphadenectomy. To improve the detection of micrometastasis in sentinel nodes of melanoma patients, molecular biological techniques were used. The exclusively melanocyte-specific tyrosinase transcript was amplified by reverse transcription followed by polymerase chain reaction (RT-PCR). In sentinel node examination, the detection of tyrosinase-positive cells by RT-PCR was compared with routine immunohistochemistry. From 16 patients, a total of 28 lymph nodes were tested. The lymph nodes were derived after lymphadenectomy (4 patients) and after sentinel node resection (12 patients with melanoma stage I). By using RT-PCR we could detect 100 tumor cells in a background of 10(8) peripheral blood mononuclear cells. The negative controls were all negative for tyrosinase. Cryostat sections of lymph nodes for mRNA isolation were alternated with sections for immunohistochemistry. By using tyrosinase RT-PCR, we detected 6 additional positive sentinel nodes in patients with melanoma stage I. Furthermore, the tyrosinase RT-PCR enabled us to design a blood test for circulating melanoma cells. Therefore, mRNA was directly isolated from whole blood of 23 blood samples, of which 3 samples were positive for tyrosinase. The present study demonstrates the possibility of a simple and rapid blood test for melanoma patients that has not been available until now. Furthermore, the detection of micrometastasis in sentinel nodes by tyrosinase RT-PCR dramatically increases the accuracy of melanoma staging.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adema G. J., de Boer A. J., Vogel A. M., Loenen W. A., Figdor C. G. Molecular characterization of the melanocyte lineage-specific antigen gp100. J Biol Chem. 1994 Aug 5;269(31):20126–20133. [PubMed] [Google Scholar]
  2. Battayani Z., Grob J. J., Xerri L., Noe C., Zarour H., Houvaeneghel G., Delpero J. R., Birmbaum D., Hassoun J., Bonerandi J. J. Polymerase chain reaction detection of circulating melanocytes as a prognostic marker in patients with melanoma. Arch Dermatol. 1995 Apr;131(4):443–447. [PubMed] [Google Scholar]
  3. Drepper H., Biess B., Hofherr B., Hundeiker M., Lippold A., Otto F., Padberg G., Peters A., Wiebelt H. The prognosis of patients with stage III melanoma. Prospective long-term study of 286 patients of the Fachklinik Hornheide. Cancer. 1993 Feb 15;71(4):1239–1246. doi: 10.1002/1097-0142(19930215)71:4<1239::aid-cncr2820710412>3.0.co;2-q. [DOI] [PubMed] [Google Scholar]
  4. Drepper H., Köhler C. O., Bastian B., Breuninger H., Bröcker E. B., Göhl J., Groth W., Hermanek P., Hohenberger W., Lippold A. Benefit of elective lymph node dissection in subgroups of melanoma patients. Results of a multicenter study of 3616 patients. Cancer. 1993 Aug 1;72(3):741–749. doi: 10.1002/1097-0142(19930801)72:3<741::aid-cncr2820720318>3.0.co;2-w. [DOI] [PubMed] [Google Scholar]
  5. Foss A. J., Guille M. J., Occleston N. L., Hykin P. G., Hungerford J. L., Lightman S. The detection of melanoma cells in peripheral blood by reverse transcription-polymerase chain reaction. Br J Cancer. 1995 Jul;72(1):155–159. doi: 10.1038/bjc.1995.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Godellas C. V., Berman C. G., Lyman G., Cruse C. W., Rapaport D., Heller R., Wang X., Glass F., Fenske N., Messina J. The identification and mapping of melanoma regional nodal metastases: minimally invasive surgery for the diagnosis of nodal metastases. Am Surg. 1995 Feb;61(2):97–101. [PubMed] [Google Scholar]
  7. Heller R., Becker J., Wasselle J., Baekey P., Cruse W., Wells K., Cox C., King B., Reintgen D. S. Detection of submicroscopic lymph node metastases in patients with melanoma. Arch Surg. 1991 Dec;126(12):1455–1460. doi: 10.1001/archsurg.1991.01410360025005. [DOI] [PubMed] [Google Scholar]
  8. Johnson P. W., Burchill S. A., Selby P. J. The molecular detection of circulating tumour cells. Br J Cancer. 1995 Aug;72(2):268–276. doi: 10.1038/bjc.1995.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Morton D. L., Wen D. R., Wong J. H., Economou J. S., Cagle L. A., Storm F. K., Foshag L. J., Cochran A. J. Technical details of intraoperative lymphatic mapping for early stage melanoma. Arch Surg. 1992 Apr;127(4):392–399. doi: 10.1001/archsurg.1992.01420040034005. [DOI] [PubMed] [Google Scholar]
  10. Reintgen D., Cruse C. W., Wells K., Berman C., Fenske N., Glass F., Schroer K., Heller R., Ross M., Lyman G. The orderly progression of melanoma nodal metastases. Ann Surg. 1994 Dec;220(6):759–767. doi: 10.1097/00000658-199412000-00009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ryan L., Kramar A., Borden E. Prognostic factors in metastatic melanoma. Cancer. 1993 May 15;71(10):2995–3005. doi: 10.1002/1097-0142(19930515)71:10<2995::aid-cncr2820711018>3.0.co;2-p. [DOI] [PubMed] [Google Scholar]
  12. Sensi M., Traversari C., Radrizzani M., Salvi S., Maccalli C., Mortarini R., Rivoltini L., Farina C., Nicolini G., Wölfel T. Cytotoxic T-lymphocyte clones from different patients display limited T-cell-receptor variable-region gene usage in HLA-A2-restricted recognition of the melanoma antigen Melan-A/MART-1. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5674–5678. doi: 10.1073/pnas.92.12.5674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Smith B., Selby P., Southgate J., Pittman K., Bradley C., Blair G. E. Detection of melanoma cells in peripheral blood by means of reverse transcriptase and polymerase chain reaction. Lancet. 1991 Nov 16;338(8777):1227–1229. doi: 10.1016/0140-6736(91)92100-g. [DOI] [PubMed] [Google Scholar]
  14. Wang X., Heller R., VanVoorhis N., Cruse C. W., Glass F., Fenske N., Berman C., Leo-Messina J., Rappaport D., Wells K. Detection of submicroscopic lymph node metastases with polymerase chain reaction in patients with malignant melanoma. Ann Surg. 1994 Dec;220(6):768–774. doi: 10.1097/00000658-199412000-00010. [DOI] [PMC free article] [PubMed] [Google Scholar]

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