Abstract
The detection of minimal amounts of melanoma cells by tyrosinase reverse transcription polymerase chain reaction (RT-PCR) is seriously hampered by false negative reports in blood of melanoma patients with disseminated melanoma. Therefore, additional assays which make use of multiple melanoma markers are needed. It has been shown that introduction of multiple markers increases the sensitivity of detection. Melanoma inhibitory activity (MIA) is one such melanoma-specific candidate gene. To test the specificity of MIA PCR, we performed 30 and 60 cycles of PCR with two different sets of MIA specific primers on 19 melanoma and 16 non-melanoma cell lines. MIA mRNA was detected in 16 out of 19 melanoma cell lines and in seven out of 16 non-melanoma cell lines after 30 cycles of PCR. However, MIA mRNA could be detected in all cell lines after 60 cycles of PCR. Also, in 14 out of 14 blood samples of melanoma patients, five out of six blood samples of non-melanoma patients and in seven out of seven blood samples of healthy volunteers, MIA mRNA was detected after 60 cycles of PCR, whereas no MIA PCR product could be detected in any of the blood samples after 30 cycles of PCR. We conclude that low levels of MIA transcripts are present in various normal and neoplastic cell types. Therefore, MIA is not a suitable marker gene to facilitate the detection of minimal amounts of melanoma cells in blood or in target organs of the metastatic process. © 1999 Cancer Research Campaign
Keywords: MIA, melanoma, circulating cancer cells, RT-PCR
Full Text
The Full Text of this article is available as a PDF (88.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Blesch A., Bosserhoff A. K., Apfel R., Behl C., Hessdoerfer B., Schmitt A., Jachimczak P., Lottspeich F., Buettner R., Bogdahn U. Cloning of a novel malignant melanoma-derived growth-regulatory protein, MIA. Cancer Res. 1994 Nov 1;54(21):5695–5701. [PubMed] [Google Scholar]
- Bosserhoff A. K., Hein R., Bogdahn U., Buettner R. Structure and promoter analysis of the gene encoding the human melanoma-inhibiting protein MIA. J Biol Chem. 1996 Jan 5;271(1):490–495. doi: 10.1074/jbc.271.1.490. [DOI] [PubMed] [Google Scholar]
- Bosserhoff A. K., Kaufmann M., Kaluza B., Bartke I., Zirngibl H., Hein R., Stolz W., Buettner R. Melanoma-inhibiting activity, a novel serum marker for progression of malignant melanoma. Cancer Res. 1997 Aug 1;57(15):3149–3153. [PubMed] [Google Scholar]
- Brossart P., Keilholz U., Willhauck M., Scheibenbogen C., Möhler T., Hunstein W. Hematogenous spread of malignant melanoma cells in different stages of disease. J Invest Dermatol. 1993 Dec;101(6):887–889. doi: 10.1111/1523-1747.ep12371713. [DOI] [PubMed] [Google Scholar]
- 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]
- Brouwenstijn N., Slager E. H., Bakker A. B., Schreurs M. W., Van der Spek C. W., Adema G. J., Schrier P. I., Figdor C. G. Transcription of the gene encoding melanoma-associated antigen gp100 in tissues and cell lines other than those of the melanocytic lineage. Br J Cancer. 1997;76(12):1562–1566. doi: 10.1038/bjc.1997.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Burchill S. A., Bradbury M. F., Pittman K., Southgate J., Smith B., Selby P. Detection of epithelial cancer cells in peripheral blood by reverse transcriptase-polymerase chain reaction. Br J Cancer. 1995 Feb;71(2):278–281. doi: 10.1038/bjc.1995.56. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Keilholz U., Willhauck M., Scheibenbogen C., de Vries T. J., Burchill S. Polymerase chain reaction detection of circulating tumour cells. EORTC Melanoma Cooperative Group, Immunotherapy Subgroup. Melanoma Res. 1997 Aug;7 (Suppl 2):S133–S141. [PubMed] [Google Scholar]
- 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]
- 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]
- Mori M., Mimori K., Tanaka F., Ueo H., Sugimachi K., Akiyoshi T. Molecular diagnosis of circulating cancer cells using MAGE gene assays. JAMA. 1997 Aug 13;278(6):476–477. [PubMed] [Google Scholar]
- 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]
- Wimmer I., Meyer J. C., Seifert B., Dummer R., Flace A., Burg G. Prognostic value of serum 5-S-cysteinyldopa for monitoring human metastatic melanoma during immunochemotherapy. Cancer Res. 1997 Nov 15;57(22):5073–5076. [PubMed] [Google Scholar]
- 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]
- de Vries T. J., Fourkour A., Wobbes T., Verkroost G., Ruiter D. J., van Muijen G. N. Heterogeneous expression of immunotherapy candidate proteins gp100, MART-1, and tyrosinase in human melanoma cell lines and in human melanocytic lesions. Cancer Res. 1997 Aug 1;57(15):3223–3229. [PubMed] [Google Scholar]
- van Groningen J. J., Bloemers H. P., Swart G. W. Identification of melanoma inhibitory activity and other differentially expressed messenger RNAs in human melanoma cell lines with different metastatic capacity by messenger RNA differential display. Cancer Res. 1995 Dec 15;55(24):6237–6243. [PubMed] [Google Scholar]
