Skip to main content
PMC home page
British Journal of Cancer logoLink to British Journal of Cancer
. 2000 Jun 15;83(2):184–187. doi: 10.1054/bjoc.2000.1282

Limitations of the nested reverse transcriptase polymerase chain reaction on tyrosinase for the detection of malignant melanoma micrometastases in lymph nodes

A Calogero 1, H Timmer-Bosscha 1, H Schraffordt Koops 2, A T M G Tiebosch 3, N H Mulder 1, G A P Hospers 1
PMCID: PMC2363472  PMID: 10901368

Abstract

The specificity and sensitivity of the nested reverse transcriptase polymerase chain reaction (RT-PCR) on tyrosinase was studied, for the detection of micrometastases of malignant melanoma. The specificity was assessed in the blood of six healthy donors, four patients with non-melanoma cancers of which one patient was treated with granulocyte-colony stimulating factor. Lymph nodes of nine patients without malignant melanoma were tested and four cell lines of various other tumours. Six of the nine non-melanoma lymph nodes were positive in this assay. The sensitivity was tested in a spike experiment in vitro, using a melanoma cell line. The detection limit was ten melanoma cells per 107peripheral blood lymphocytes. © 2000 Cancer Research Campaign

Keywords: tyrosinase, melanoma, lymph nodes, nested RT-PCR

Full Text

The Full Text of this article is available as a PDF (78.5 KB).

Selected References

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

  1. Andrews P. W., Damjanov I., Simon D., Banting G. S., Carlin C., Dracopoli N. C., Føgh J. Pluripotent embryonal carcinoma clones derived from the human teratocarcinoma cell line Tera-2. Differentiation in vivo and in vitro. Lab Invest. 1984 Feb;50(2):147–162. [PubMed] [Google Scholar]
  2. Battyani Z., Xerri L., Hassoun J., Bonerandi J. J., Grob J. J. Tyrosinase gene expression in human tissues. Pigment Cell Res. 1993 Dec;6(6):400–405. doi: 10.1111/j.1600-0749.1993.tb00622.x. [DOI] [PubMed] [Google Scholar]
  3. Bautista N. C., Cohen S., Anders K. H. Benign melanocytic nevus cells in axillary lymph nodes. A prospective incidence and immunohistochemical study with literature review. Am J Clin Pathol. 1994 Jul;102(1):102–108. doi: 10.1093/ajcp/102.1.102. [DOI] [PubMed] [Google Scholar]
  4. Berendsen H. H., de Leij L., de Vries E. G., Mesander G., Mulder N. H., de Jong B., Buys C. H., Postmus P. E., Poppema S., Sluiter H. J. Characterization of three small cell lung cancer cell lines established from one patient during longitudinal follow-up. Cancer Res. 1988 Dec 1;48(23):6891–6899. [PubMed] [Google Scholar]
  5. Bieligk S. C., Ghossein R., Bhattacharya S., Coit D. G. Detection of tyrosinase mRNA by reverse transcription-polymerase chain reaction in melanoma sentinel nodes. Ann Surg Oncol. 1999 Apr-May;6(3):232–240. doi: 10.1007/s10434-999-0232-z. [DOI] [PubMed] [Google Scholar]
  6. Blaheta H. J., Schittek B., Breuninger H., Maczey E., Kroeber S., Sotlar K., Ellwanger U., Thelen M. H., Rassner G., Bültmann B. Lymph node micrometastases of cutaneous melanoma: increased sensitivity of molecular diagnosis in comparison to immunohistochemistry. Int J Cancer. 1998 Aug 21;79(4):318–323. doi: 10.1002/(sici)1097-0215(19980821)79:4<318::aid-ijc3>3.0.co;2-y. [DOI] [PubMed] [Google Scholar]
  7. Brossart P., Schmier J. W., Krüger S., Willhauck M., Scheibenbogen C., Möhler T., Keilholz U. A polymerase chain reaction-based semiquantitative assessment of malignant melanoma cells in peripheral blood. Cancer Res. 1995 Sep 15;55(18):4065–4068. [PubMed] [Google Scholar]
  8. Chelly J., Concordet J. P., Kaplan J. C., Kahn A. Illegitimate transcription: transcription of any gene in any cell type. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2617–2621. doi: 10.1073/pnas.86.8.2617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Curry B. J., Myers K., Hersey P. Polymerase chain reaction detection of melanoma cells in the circulation: relation to clinical stage, surgical treatment, and recurrence from melanoma. J Clin Oncol. 1998 May;16(5):1760–1769. doi: 10.1200/JCO.1998.16.5.1760. [DOI] [PubMed] [Google Scholar]
  10. Farthmann B., Eberle J., Krasagakis K., Gstöttner M., Wang N., Bisson S., Orfanos C. E. RT-PCR for tyrosinase-mRNA-positive cells in peripheral blood: evaluation strategy and correlation with known prognostic markers in 123 melanoma patients. J Invest Dermatol. 1998 Mar;110(3):263–267. doi: 10.1046/j.1523-1747.1998.00131.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Ghossein R. A., Coit D., Brennan M., Zhang Z. F., Wang Y., Bhattacharya S., Houghton A., Rosai J. Prognostic significance of peripheral blood and bone marrow tyrosinase messenger RNA in malignant melanoma. Clin Cancer Res. 1998 Feb;4(2):419–428. [PubMed] [Google Scholar]
  13. Gläser R., Rass K., Seiter S., Hauschild A., Christophers E., Tilgen W. Detection of circulating melanoma cells by specific amplification of tyrosinase complementary DNA is not a reliable tumor marker in melanoma patients: a clinical two-center study. J Clin Oncol. 1997 Aug;15(8):2818–2825. doi: 10.1200/JCO.1997.15.8.2818. [DOI] [PubMed] [Google Scholar]
  14. Goydos J. S., Ravikumar T. S., Germino F. J., Yudd A., Bancila E. Minimally invasive staging of patients with melanoma: sentinel lymphadenectomy and detection of the melanoma-specific proteins MART-1 and tyrosinase by reverse transcriptase polymerase chain reaction. J Am Coll Surg. 1998 Aug;187(2):182–190. doi: 10.1016/s1072-7515(98)00138-0. [DOI] [PubMed] [Google Scholar]
  15. Hatta N., Takata M., Takehara K., Ohara K. Polymerase chain reaction and immunohistochemistry frequently detect occult melanoma cells in regional lymph nodes of melanoma patients. J Clin Pathol. 1998 Aug;51(8):597–601. doi: 10.1136/jcp.51.8.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hoon D. S., Wang Y., Dale P. S., Conrad A. J., Schmid P., Garrison D., Kuo C., Foshag L. J., Nizze A. J., Morton D. L. Detection of occult melanoma cells in blood with a multiple-marker polymerase chain reaction assay. J Clin Oncol. 1995 Aug;13(8):2109–2116. doi: 10.1200/JCO.1995.13.8.2109. [DOI] [PubMed] [Google Scholar]
  17. Jung F. A., Buzaid A. C., Ross M. I., Woods K. V., Lee J. J., Albitar M., Grimm E. A. Evaluation of tyrosinase mRNA as a tumor marker in the blood of melanoma patients. J Clin Oncol. 1997 Aug;15(8):2826–2831. doi: 10.1200/JCO.1997.15.8.2826. [DOI] [PubMed] [Google Scholar]
  18. Kunter U., Buer J., Probst M., Duensing S., Dallmann I., Grosse J., Kirchner H., Schluepen E. M., Volkenandt M., Ganser A. Peripheral blood tyrosinase messenger RNA detection and survival in malignant melanoma. J Natl Cancer Inst. 1996 May 1;88(9):590–594. doi: 10.1093/jnci/88.9.590. [DOI] [PubMed] [Google Scholar]
  19. Leibovitz A., Stinson J. C., McCombs W. B., 3rd, McCoy C. E., Mazur K. C., Mabry N. D. Classification of human colorectal adenocarcinoma cell lines. Cancer Res. 1976 Dec;36(12):4562–4569. [PubMed] [Google Scholar]
  20. Mellado B., Colomer D., Castel T., Muñoz M., Carballo E., Galán M., Mascaró J. M., Vives-Corrons J. L., Grau J. J., Estapé J. Detection of circulating neoplastic cells by reverse-transcriptase polymerase chain reaction in malignant melanoma: association with clinical stage and prognosis. J Clin Oncol. 1996 Jul;14(7):2091–2097. doi: 10.1200/JCO.1996.14.7.2091. [DOI] [PubMed] [Google Scholar]
  21. Palmieri G., Strazzullo M., Ascierto P. A., Satriano S. M., Daponte A., Castello G. Polymerase chain reaction-based detection of circulating melanoma cells as an effective marker of tumor progression. Melanoma Cooperative Group. J Clin Oncol. 1999 Jan;17(1):304–311. doi: 10.1200/JCO.1999.17.1.304. [DOI] [PubMed] [Google Scholar]
  22. Rankin E. M. Detection of early micrometastases in malignant melanoma. Eur J Cancer. 1996 Sep;32A(10):1627–1629. doi: 10.1016/0959-8049(96)00182-7. [DOI] [PubMed] [Google Scholar]
  23. Reinhold U., Lüdtke-Handjery H. C., Schnautz S., Kreysel H. W., Abken H. The analysis of tyrosinase-specific mRNA in blood samples of melanoma patients by RT-PCR is not a useful test for metastatic tumor progression. J Invest Dermatol. 1997 Feb;108(2):166–169. doi: 10.1111/1523-1747.ep12333341. [DOI] [PubMed] [Google Scholar]
  24. Rogan A. M., Hamilton T. C., Young R. C., Klecker R. W., Jr, Ozols R. F. Reversal of adriamycin resistance by verapamil in human ovarian cancer. Science. 1984 Jun 1;224(4652):994–996. doi: 10.1126/science.6372095. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Van der Velde-Zimmermann D., Roijers J. F., Bouwens-Rombouts A., De Weger R. A., De Graaf P. W., Tilanus M. G., Van den Tweel J. G. Molecular test for the detection of tumor cells in blood and sentinel nodes of melanoma patients. Am J Pathol. 1996 Sep;149(3):759–764. [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. de Vries T. J., Fourkour A., Punt C. J., van de Locht L. T., Wobbes T., van den Bosch S., de Rooij M. J., Mensink E. J., Ruiter D. J., van Muijen G. N. Reproducibility of detection of tyrosinase and MART-1 transcripts in the peripheral blood of melanoma patients: a quality control study using real-time quantitative RT-PCR. Br J Cancer. 1999 May;80(5-6):883–891. doi: 10.1038/sj.bjc.6690436. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES