Molekularbiologischer Nachweis von Tumormarkern im peripheren Blut und Plasma/Serum von Patienten mit malignem Melanom
Molecular Detection of Tumor Markers in Peripheral Blood and Plasma/Serum of Patients with Malignant MelanomaG. Rappl1
, S. Ugurel1
, W. Tilgen1
, U. Reinhold1
1Universitäts-Hautklinik und Poliklinik, Universitätskliniken des Saarlandes, Homburg/Saar
Der frühe Nachweis einer Metastasierung beim malignen Melanom ist von entscheidender Bedeutung für die Prognose der Erkrankung. Zum Nachweis zirkulierender Tumorzellen wurden im Laufe der Jahre zunehmend sensitivere Methoden eingesetzt. Durch die Entwicklung der revers-transkribierten Polymerasekettenreaktion (RT-PCR) wurde ein spezifischer und sensitiver Nachweis zirkulierender Tumorzellen möglich. Beim malignen Melanom wurden mittels RT-PCR melanomspezifische Transkripte, insbesondere die Expression des Genes für das Enzym Tyrosinase in umfangreichen Studien untersucht. Weiterführende Projekte untersuchten die Expression weiterer Melanom-assoziierter Transkripte („Multimarker RT-PCR”), sowie den Einsatz der quantitativen „real-time”-RT-PCR zur Optimierung der Sensitivität und Spezifität zur Detektion von Mikrometastasen. Einen weiteren noch in der experimentellen Phase befindlichen Ansatz zum Nachweis von Mikrometastasen stellt die Analyse von frei zirkulierenden tumorassoziierten Nukleinsäuren im Plasma/Serum dar. Erste Daten bestätigen die prinzipielle Möglichkeit dieser Nachweismethodik als neuen Ansatz für den Nachweis einer Mikrometastasierung.
Abstract
The early detection of metastasis in malignant melanoma is important for prognosis of disease. For detection of circulating tumor cells more sensitive methods were developed during past years. Through the development of polymerase-chain-reaction a specific and sensitive detection of circulating tumor cells is possible. The expression of melanoma-associated transcripts in malignant melanoma, especially the expression of the tyrosinase gene, was analyzed in multiple studies. More recent studies analyze expression of different melanoma-associated transcripts (multi-marker RT-PCR) or usage of real-time RT-PCR for optimization of detection sensitivity and specificity of micrometastases. A new method under development is the detection of tumor-associated nucleic acids in plasma/serum. First data confirm the possible usage of this method for detection of micrometastases.
Literatur
1 Garbe C, Schaumburg-Lever G. Klinik und Histologie des malignen Melanoms. In: Garbe C, Dummer R, Kaufmann R, Tilgen W (Hrsg) Dermatologische Onkologie. Berlin Heidelberg New York Tokyo; Springer 1997
2
Heimdal K, Hannisdal E, Gundersen S.
Regression analysis of prognostic factors in metastatic malignant melanoma.
Eur J Cancer Clin Oncol.
1989;
25
1219-1223
3
Henze G, Dummer R, Joller-Jemelka H, Böni R, Burg G.
Serum S100, a marker for disease monitoring in metastatic melanoma.
Dermatology.
1997;
194
208-212
4
Pelkey T J, Frierson H F, Bruns D E.
Molecular and immunological detection of circulating tumor cells and micrometastasis from solid tumors.
Clin Chem.
1996;
42
1369-1381
5
Miettinen M.
Keratin subsets in spindle cell sarcomas. Keratins are widespread but synovial sarcoma contains a distinctive keratin polypeptide pattern and desmoplakins.
Am J Pathol.
1991;
138
505-513
6
Thomas P, Battifora H.
Keratins versus epithelial membrane antigen in tumor diagnosis- an immunohistochemical comparison of five monoclonal antibodies.
Hum Pathol.
1987;
18
728-734
7
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;
338
1227-1229
8
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
887-889
9 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. In: Reinhold U, Tilgen W (Hrsg) Minimal residual disease in melanoma. Recent Results in Cancer Res. Vol. 158. Berlin Heidelberg New York Tokyo; Springer 2000
10
Reinhold U, Berkin C, Bosserhoff A K, Deutschmann A, Garbe C, Gläser R, Hein R, Krahn G, Peter R U, Rappl G, Schittek B, Seiter S, Ugurel S, Volkenandt M, Tilgen W.
Interlaboratory evaluation of a new reverse transcriptase polmerase chain reaction-based enzyme-linked immunosorbent assay for the detection of circulating tumor cells: A multicenter study of the dermatologic cooperative oncology group.
J Clin Oncol.
2001;
19
1723-1727
11
Marincola F M, Jaffeee E M, Hicklin D J, Ferrone S.
Escape of solid human tumors from T-cell recognition: molecular mechanisms and functional significance.
Adv Immunol.
2000;
74
181-273
12
Hoon D SB, 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;
13
2109-2116
13
Sarantou T, Chi D D, Garrison D A, Conrad A J, Schmid P, Morton D L, Hoon D SB.
Melanoma-associated antigens as messenger RNA detection markers for melanoma.
Cancer Res..
1997;
57
1371-1376
14 Taback B, Morton D L, O'Day S J, Nguyen D H, Nakayama T, Hoon D SB. The clinical utility of multimarker RT-PCR in the detection of occult metastasis in patients with melanoma. In: Reinhold U, Tilgen W (Hrsg) Minimal residual disease in melanoma. Recent Results in Cancer Res. Vol. 158. Berlin Heidelberg New York Tokyo; Springer 2000
15
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;
80
883-891
16 Max N, Wolf K, Spike E, Thiel E, Keilholz U. Nested quantitative real time PCR for detection of occult tumor cells. In: Reinhold U, Tilgen W (Hrsg) Minimal residual disease in melanoma. Recent Results in Cancer Res. Vol. 158. Berlin Heidelberg New York Tokyo; Springer 2000
17
Kopreski M S, Benko F A, Kwak L W, Gocke C D.
Detection of tumor messenger mRNA in the serum of patients with malignant melanoma.
Clin Cancer Res.
1999;
5
1961-1965
18
Hasselmann D O, Rappl G, Rößler M, Ugurel S, Tilgen W, Reinhold U.
Detection of tumor-associated circulating mRNA in serum, plasma and blood cell samples from patients with disseminated malignant melanoma.
Oncol Rep.
2001;
8
115-118
20
Wieczorek A J, Rhyner C, Block L H.
Isolation and characterization of an RNA-proteolipid complex associated with the malignant state in humans.
Proc Natl Acad Sci USA.
1985;
82
3455-3459
21
Hasselmann D O, Rappl G, Tilgen W, Reinhold U.
Extracellular tyrosinase mRNA within apoptotic bodies is protected from degradation in human serum.
Clin Chem.
2001;
47
1488-1489
22
Jahr S, Hentze H, Englisch S, Hardt D, Fackelmayer F O, Hesch R D, Knippers R.
DNA fragments in the blood of cancer patients: quantitations and evidence for their origin from apoptotic and necrotic cells.
Cancer Res.
2001;
61
1659-1665
24
Silva J M, Dominguez G, Silva J, Garcia J M, Sanchez A, Rodriguez O, Provencio M, Espana P, Bonilla F.
Detection of epithelial messenger RNA in the plasma of breast cancer patients is associated with poor prognosis tumor characteristics.
Clin Cancer Res.
2001;
7
2821-2825