Startseite Medizin Molecular detection of tyrosinase transcripts in peripheral blood from patients with malignant melanoma: correlation of PCR sensitivity threshold with clinical and pathologic disease characteristics
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Molecular detection of tyrosinase transcripts in peripheral blood from patients with malignant melanoma: correlation of PCR sensitivity threshold with clinical and pathologic disease characteristics

  • Georgios Mitropapas , Andrianos Nezos , Antonis Halapas , Nikolaos Pissimissis , Peter Lembessis , Antigone Sourla , Pericles Vassilopoulos und Michael Koutsilieris
Veröffentlicht/Copyright: 13. Dezember 2006

Abstract

Background: Positive molecular detection of tyrosinase transcripts (TYR mRNA) in RNA extracts of peripheral blood (PB) samples from patients with malignant melanoma provides evidence of disease dissemination.

Methods: Total RNA extracted from PB was quantified and subjected to RT-PCR under ultra-sensitive and reduced-sensitivity PCR conditions using SSRT-II. Positive TYR mRNA detection in 78 melanoma patients and 40 healthy volunteers was correlated with clinical stage, Breslow's evaluation of tumor thickness, Clark's assessment of tumor invasion, the location of the primary tumor site, and tumor histology. The assay sensitivity was evaluated by spiking PB with the melanoma cell line SK-MEL-28.

Results: Using ultra-sensitive PCR conditions, eight out of 40 RNA (20%) samples from healthy volunteers and 50 out of 78 RNA (64.1%) samples from melanoma patients tested positive. Using reduced-sensitivity PCR conditions, we found only two positives in 40 RNA samples from healthy subjects and 20 positives in 78 RNA samples (25.6%) from melanoma patients. Only positive PCR samples for the reduced-sensitivity PCR assay correlated significantly with stage IV (metastatic) disease (p=0.0395). There was no significant correlation between positive TYR mRNA samples for either PCR condition (ultra-sensitive and reduced-sensitivity) with Breslow's classification of tumor thickness, Clark's assessment of tumor invasion, location of the primary tumor site, and type of tumor histology.

Conclusions: We conclude that reduced-sensitivity rather than ultra-sensitive PCR conditions correlate with clinical stage in melanoma patients.

Clin Chem Lab Med 2006;44:1403–9.


Corresponding author: Michael Koutsilieris, MD, PhD, Professor of Experimental Physiology and Chairman of the Departments of Basic Sciences, Medical School, University of Athens, Micras Asias 75, Goudi, 11527, Athens, Greece Phone: +30-210-7462597, Fax: +30-210-74625761,

References

1. Farthmann B, Eberle J, Krasagakis K, Gstottner M, Wang N, Bisson S, et al. 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; 110:263–7.10.1046/j.1523-1747.1998.00131.xSuche in Google Scholar PubMed

2. Palmieri G, Strazzullo M, Ascierto PA, Satriano SM, 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; 17:304–11.10.1200/JCO.1999.17.1.304Suche in Google Scholar PubMed

3. Blaheta HJ, Paul T, Sotlar K, Maczey E, Schittek B, Paul A, et al. Detection of melanoma cells in sentinel lymph nodes, bone marrow and peripheral blood by a reverse transcription-polymerase chain reaction assay in patients with primary cutaneous melanoma: association with Breslow's tumour thickness. Br J Dermatol 2001; 145:195–202.10.1046/j.1365-2133.2001.04334.xSuche in Google Scholar PubMed

4. Battayani Z, Grob JJ, Xerri L, Noe C, Zarour H, Houvaeneghel G, et al. Polymerase chain reaction detection of circulating melanocytes as a prognostic marker in patients with melanoma. Arch Dermatol 1995; 131:443–7.10.1001/archderm.1995.01690160071011Suche in Google Scholar

5. Seiter S, Rappl G, Tilgen W, Ugurel S, Reinhold U. Facts and pitfalls in the detection of tyrosinase mRNA in the blood of melanoma patients by RT-PCR. Recent Results Cancer Res 2001; 158:105–12.10.1007/978-3-642-59537-0_10Suche in Google Scholar PubMed

6. Reinhold U, Berkin C, Bosserhoff AK, Deutschmann A, Garbe C, Glaser R, et al. Interlaboratory evaluation of a new reverse transcriptase polymerase chain reaction-based enzyme-linked immunosorbent assay for the detection of circulating melanoma cells: a multicenter study of the Dermatologic Cooperative Oncology Group. J Clin Oncol 2001; 19:1723–7.10.1200/JCO.2001.19.6.1723Suche in Google Scholar PubMed

7. Pittman K, Burchill S, Smith B, Southgate J, Joffe J, Gore M, et al. Reverse transcriptase-polymerase chain reaction for expression of tyrosinase to identify malignant melanoma cells in peripheral blood. Ann Oncol 1996; 7:297–301.10.1093/oxfordjournals.annonc.a010575Suche in Google Scholar PubMed

8. Ghossein RA, Coit D, Brennan M, Zhang ZF, Wang Y, Bhattacharya S, et al. Prognostic significance of peripheral blood and bone marrow tyrosinase messenger RNA in malignant melanoma. Clin Cancer Res 1998; 4:419–28.Suche in Google Scholar

9. Kwon BS, Halaban R, Kim GS, Usack L, Pomerantz S, Haq AK. A melanocyte-specific complementary DNA clone whose expression is inducible by melanotropin and isobutylmethyl xanthine. Mol Biol Med 1987; 4:339–55.Suche in Google Scholar

10. Kunter U, Buer J, Probst M, Duensing S, Dallmann I, Grosse J, et al. Peripheral blood tyrosinase messenger RNA detection and survival in malignant melanoma. J Natl Cancer Inst 1996; 88:590–4.10.1093/jnci/88.9.590Suche in Google Scholar PubMed

11. Zimmermann H, Eggers HJ, Nelsen-Salz B. Molecular cloning and sequence determination of the complete genome of the virulent echovirus 9 strain barty. Virus Genes 1996; 12:149–54.10.1007/BF00572953Suche in Google Scholar PubMed

12. Calogero A, Hospers GA, Timmer-Bosscha H, Koops HS, Mulder NH. Effect of specific or random c-DNA priming on sensitivity of tyrosinase nested RT-PCR: potential clinical relevance. Anticancer Res 2000; 20:3545–8.Suche in Google Scholar

13. Proebstle TM, Jiang W, Hogel J, Keilholz U, Weber L, Voit C. Correlation of positive RT-PCR for tyrosinase in peripheral blood of malignant melanoma patients with clinical stage, survival and other risk factors. Br J Cancer 2000; 82:118–23.10.1054/bjoc.1998.0887Suche in Google Scholar PubMed PubMed Central

14. Sardi I, Moretti S, Ponchietti R, Arrigucci S, Guazzelli R, Montali E. The role of the detection of hematogenous micrometastasis in prostate adenocarcinoma and malignant melanoma by RT-PCR. Int J Mol Med 1999; 3:417–9.10.3892/ijmm.3.4.417Suche in Google Scholar

15. Hasselmann DO, Rappl G, Rossler M, Ugurel S, Tilgen W, Reinhold U. Detection of tumor-associated circulating mRNA in serum, plasma and blood cells from patients with disseminated malignant melanoma. Oncol Rep 2001; 8:115–8.10.3892/or.8.1.115Suche in Google Scholar

16. Reinhold U, Ludtke-Handjery HC, Schnautz S, Kreysel HW, 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; 108:166–9.10.1111/1523-1747.ep12333341Suche in Google Scholar

17. Brossart P, Keilholz U, Willhauck M, Scheibenbogen C, Mohler T, Hunstein W. Hematogenous spread of malignant melanoma cells in different stages of disease. J Invest Dermatol 1993; 101:887–9.10.1111/1523-1747.ep12371713Suche in Google Scholar

18. Schittek B, Blaheta HJ, Florchinger G, Sauer B, Garbe C. Increased sensitivity for the detection of malignant melanoma cells in peripheral blood using an improved protocol for reverse transcription-polymerase chain reaction. Br J Dermatol 1999; 141:37–43.10.1046/j.1365-2133.1999.02918.xSuche in Google Scholar

19. Foss AJ, Guille MJ, Occleston NL, Hykin PG, Hungerford JL, Lightman S. The detection of melanoma cells in peripheral blood by reverse transcription-polymerase chain reaction. Br J Cancer 1995; 72:155–9.10.1038/bjc.1995.293Suche in Google Scholar

20. Mellado B, Colomer D, Castel T, Munoz M, Carballo E, Galan M, 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; 14:2091–7.10.1200/JCO.1996.14.7.2091Suche in Google Scholar

21. Keilholz U. Diagnostic PCR in melanoma, methods and quality assurance. Epalinges, Switzerland, 26/27 January 1996. Eur J Cancer 1996; 32A:1661–3.10.1016/0959-8049(96)00158-XSuche in Google Scholar

22. Schittek B, Blaheta HJ, Ellwanger U, Garbe C. Polymerase chain reaction in the detection of circulating tumour cells in peripheral blood of melanoma patients. Recent Results Cancer Res 2001; 158:93–104.10.1007/978-3-642-59537-0_9Suche in Google Scholar

23. Tsao H, Nadiminti U, Sober AJ, Bigby M. A meta-analysis of reverse transcriptase-polymerase chain reaction for tyrosinase mRNA as a marker for circulating tumor cells in cutaneous melanoma. Arch Dermatol 2001; 137:325–30.Suche in Google Scholar

24. Keilholz U, Willhauck M, Rimoldi D, Brasseur F, Dummer W, Rass K, et al. Reliability of reverse transcription-polymerase chain reaction (RT-PCR)-based assays for the detection of circulating tumour cells: a quality-assurance initiative of the EORTC melanoma cooperative group. Eur J Cancer 1998; 34:750–3.10.1016/S0959-8049(97)10105-8Suche in Google Scholar

25. Santonocito C, Concolino P, Lavieri MM, Ameglio F, Gentileschi S, Capizzi R, et al. Comparison between three molecular methods for detection of blood melanoma tyrosinase mRNA. Correlation with melanoma stages and S100B, LDH, NSE biochemical markers. Clin Chim Acta 2005; 362:85–93.10.1016/j.cccn.2005.05.031Suche in Google Scholar

26. Szenajch J, Jasinski B, Synowiec A, Kulik J, Chomicka M, Struzyna J, et al. Prognostic value of multiple reverse transcription-PCR tyrosinase testing for circulating neoplastic cells in malignant melanoma. Clin Chem 2003; 49:1450–7.10.1373/49.9.1450Suche in Google Scholar

27. Warr RP, Zebedeeb Z, Kenealya J, Rigbyc H, Kemsheadb JT. The detection of tyrosinase mRNA in peripheral blood samples is unlikely to aid in the management of patients with localised malignant melanoma. Br Assoc Plast Surg 2003; 56:540–5.10.1016/S0007-1226(03)00189-9Suche in Google Scholar

28. Prichard RS, Dijkstra B, McDermott EW, Hill AD, O'Higgins NJ. The role of molecular staging in malignant melanoma. EJSO 2003; 29:306–14.10.1053/ejso.2002.1366Suche in Google Scholar

29. 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; 88:569–70.10.1093/jnci/88.9.569Suche in Google Scholar

30. Jin H-Y, Yamashita T, Minamitsuji Y, Omori F, Jimbow K. Detection of tyrosinase and tyrosinase-related protein 1 sequences from peripheral blood of melanoma patients using reverse transcription polymerase chain reaction. J Dermatol Sci 2003; 33:169–76.10.1016/j.jdermsci.2003.09.003Suche in Google Scholar

31. Fidler IJ. Critical factors in the biology of human cancer metastasis: 28th GHA Clowes memorial award lecture. Cancer Res 1990; 50:6130–8.Suche in Google Scholar

32. Liotta LA, Kleinerman J, Saidel GM. Quantitative relationships of intravascular tumor cells, tumor vessels, and pulmonary metastases following tumor implantation. Cancer Res 1974; 34:997–1004.Suche in Google Scholar

33. Gomella LG, Raj GV, Moreno JG. Reverse transcriptase polymerase chain reaction for prostate specific antigen in the management of prostate cancer. J Urol 1997; 158:326–37.10.1016/S0022-5347(01)64472-0Suche in Google Scholar

34. Baldi A, Dragonetti E, Battista T, Groeger AM, Esposito V, Baldi G, et al. Detection of circulating malignant cells by RT-PCR in long-term clinically disease-free I stage melanoma patients. Anticancer Res 2000; 20:3923–8.Suche in Google Scholar

35. Russell-Jones R. The value of reverse transcription-polymerase chain reaction in malignant melanoma. Br J Dermatol 2001; 145:193–4.10.1046/j.1365-2133.2001.04332.xSuche in Google Scholar

36. Brownbridge GG, Gold J, Edward M, MacKie RM. Evaluation of the use of tyrosinase-specific and melanA/MART-1-specific reverse transcriptase-coupled--polymerase chain reaction to detect melanoma cells in peripheral blood samples from 299 patients with malignant melanoma. Br J Dermatol 2001; 144:279–87.10.1046/j.1365-2133.2001.04015.xSuche in Google Scholar

37. Le Bricon T, Stoitchkov K, Letellier S, Guibal F, Spy J, Garnier JP, et al. Simultaneous analysis of tyrosinase mRNA and markers of tyrosinase activity in the blood of patients with metastatic melanoma. Clin Chim Acta 1999; 282:101–13.10.1016/S0009-8981(99)00022-4Suche in Google Scholar

38. Mellado B, Gutierrez L, Castel T, Colomer D, Fontanillas M, Castro J, et al. Prognostic significance of the detection of circulating malignant cells by reverse transcriptase-polymerase chain reaction in long-term clinically disease-free melanoma patients. Clin Cancer Res 1999; 5:1843–8.Suche in Google Scholar

39. Quaglino P, Savoia P, Fierro MT, Osella-Abate S, Bernengo MG. Clinical significance of sequential tyrosinase expression in the peripheral blood of disease-free melanoma patients: a review of literature data. Melanoma Res 2004; 14(Suppl 1):S17–9.10.1097/00008390-200404000-00018Suche in Google Scholar PubMed

40. Palmieri G, Ascierto PA, Perrone F, Satriano SM, Ottaiano A, Daponte A, et al. Prognostic value of circulating melanoma cells detected by reverse transcriptase-polymerase chain reaction. J Clin Oncol 2003; 21:767–73.10.1200/JCO.2003.01.128Suche in Google Scholar PubMed

41. Šamija I, Lukaè J, Mariæ-Broziæ J, Kusiæ Z. Microphthalmia-associated transcription factor and tyrosinase as markers of melanoma cells in blood of patients with melanoma. Croat Med J 2004; 45:142–8.Suche in Google Scholar

42. Taback B, Morton DL, O'Day SJ, Nguyen DH, Nakayama T, Hoon DS. The clinical utility of multimarker RT-PCR in the detection of occult metastasis in patients with melanoma [review]. Recent Results Cancer Res 2001; 158:78–92.10.1007/978-3-642-59537-0_8Suche in Google Scholar PubMed

43. Hoon DS, Bostick P, Kuo C, Okamoto T, Wang HJ, Elashoff R, Morton DL. Molecular markers in blood as surrogate prognostic indicators of melanoma recurrence. Cancer Res 2000; 60:2253–7.Suche in Google Scholar

44. Enk Claes D, Lotem M, Gimon Z, Hochberg M. Molecular detection of MART-1, tyrosinase and MIA in peripheral blood, lymph nodes and metastatic sites of stage III/IV melanoma patients. Melanoma Res 2004; 14:361–5.10.1097/00008390-200410000-00005Suche in Google Scholar PubMed

45. Davids V, Kidson SH, Hanekom GS. Accurate molecular detection of melanoma nodal metastases: an assessment of multimarker assay specificity, sensitivity, and detection rate. Clin Path Mol Pathol 2004; 56:43–51.10.1136/mp.56.1.43Suche in Google Scholar PubMed PubMed Central

Received: 2006-6-9
Accepted: 2006-9-12
Published Online: 2006-12-13
Published in Print: 2006-12-1

©2006 by Walter de Gruyter Berlin New York

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