The detection of melanoma cells in peripheral blood by reverse transcription-polymerase chain reaction (original) (raw)

Abstract

Both cutaneous and uveal melanoma undergo haematogenous dissemination. Detection of tyrosinase mRNA by reverse transcription-polymerase chain reaction (RT-PCR) has been described as an extremely sensitive way of detecting circulating viable melanoma cells in the peripheral venous blood, and this technique may be of value in the early detection of dissemination. Also, it has been suggested that surgical manipulation of the eye, such as occurs during enucleation, can provoke uveal melanoma dissemination. The purpose of this study was to evaluate whether tyrosinase mRNA is detectable in the peripheral blood of patients with uveal and cutaneous melanoma and in patients with uveal melanoma undergoing surgical procedures on the eye harbouring the tumour. Venous blood samples from 36 patients diagnosed as having active uveal melanoma and from six patients with advanced metastatic cutaneous melanoma were analysed. In addition, blood samples were spiked with known numbers of cells from three cell lines and four primary uveal melanoma cultures. The reported sensitivity of the technique was confirmed, with an ability to detect down to one cell per ml of blood. All 51 blood samples from the 36 patients with uveal melanoma were negative, and this included 20 perioperative blood samples. The test was also negative for the six patients with advanced cutaneous melanoma. There were two positives among 31 control samples analysed. This study demonstrates that there are far fewer circulating viable melanocytes than has been previously supposed in patients with melanoma and that the RT-PCR is of no clinical value in detecting metastatic melanoma disease. There was no evidence for surgery causing a bolus of melanoma cells to enter the peripheral circulation.

155

Images in this article

Selected References

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

  1. Augsburger J. J., Gamel J. W., Lauritzen K., Brady L. W. Cobalt-60 plaque radiotherapy vs enucleation for posterior uveal melanoma. Am J Ophthalmol. 1990 May 15;109(5):585–592. doi: 10.1016/s0002-9394(14)70691-9. [DOI] [PubMed] [Google Scholar]
  2. Burchill S. A., Bradbury F. M., Smith B., Lewis I. J., Selby P. Neuroblastoma cell detection by reverse transcriptase-polymerase chain reaction (RT-PCR) for tyrosine hydroxylase mRNA. Int J Cancer. 1994 Jun 1;57(5):671–675. doi: 10.1002/ijc.2910570510. [DOI] [PubMed] [Google Scholar]
  3. Char D. H. Metastatic choroidal melanoma. Am J Ophthalmol. 1978 Jul;86(1):76–80. doi: 10.1016/0002-9394(78)90018-1. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Foulds W. S., Damato B. E., Burton R. L. Local resection versus enucleation in the management of choroidal melanoma. Eye (Lond) 1987;1(Pt 6):676–679. doi: 10.1038/eye.1987.110. [DOI] [PubMed] [Google Scholar]
  7. Fraunfelder F. T., Boozman F. W., Wilson R. S., Thomas A. H. No-touch technique for intraocular malignant melanomas. Arch Ophthalmol. 1977 Sep;95(9):1616–1620. doi: 10.1001/archopht.1977.04450090138012. [DOI] [PubMed] [Google Scholar]
  8. GOLDBLATT S. A., NADEL E. M. CANCER CELLS IN THE CIRCULATING BLOOD: A CRITICAL REVIEW II. Acta Cytol. 1965 Jan-Feb;9:6–20. [PubMed] [Google Scholar]
  9. Kwon B. S., Haq A. K., Pomerantz S. H., Halaban R. Isolation and sequence of a cDNA clone for human tyrosinase that maps at the mouse c-albino locus. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7473–7477. doi: 10.1073/pnas.84.21.7473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lo Y. M., Patel P., Wainscoat J. S., Sampietro M., Gillmer M. D., Fleming K. A. Prenatal sex determination by DNA amplification from maternal peripheral blood. Lancet. 1989 Dec 9;2(8676):1363–1365. doi: 10.1016/s0140-6736(89)91969-7. [DOI] [PubMed] [Google Scholar]
  11. MCGREW E. A. CRITERIA FOR THE RECOGNITION OF MALIGNANT CELLS IN CIRCULATING BLOOD. Acta Cytol. 1965 Jan-Feb;9:58–60. [PubMed] [Google Scholar]
  12. McLean I. W., Foster W. D., Zimmerman L. E., Martin D. G. Inferred natural history of uveal melanoma. Invest Ophthalmol Vis Sci. 1980 Jul;19(7):760–770. [PubMed] [Google Scholar]
  13. McLean I. W., Foster W. D., Zimmerman L. E. Uveal melanoma: location, size, cell type, and enucleation as risk factors in metastasis. Hum Pathol. 1982 Feb;13(2):123–132. doi: 10.1016/s0046-8177(82)80116-0. [DOI] [PubMed] [Google Scholar]
  14. McLean M. J., Foster W. D., Zimmerman L. E. Prognostic factors in small malignant melanomas of choroid and ciliary body. Arch Ophthalmol. 1977 Jan;95(1):48–58. doi: 10.1001/archopht.1977.04450010050004. [DOI] [PubMed] [Google Scholar]
  15. Moss T. J., Sanders D. G. Detection of neuroblastoma cells in blood. J Clin Oncol. 1990 Apr;8(4):736–740. doi: 10.1200/JCO.1990.8.4.736. [DOI] [PubMed] [Google Scholar]
  16. Redding W. H., Coombes R. C., Monaghan P., Clink H. M., Imrie S. F., Dearnaley D. P., Ormerod M. G., Sloane J. P., Gazet J. C., Powles T. J. Detection of micrometastases in patients with primary breast cancer. Lancet. 1983 Dec 3;2(8362):1271–1274. doi: 10.1016/s0140-6736(83)91150-9. [DOI] [PubMed] [Google Scholar]
  17. SEAL S. H. A SIEVE FOR THE ISOLATION OF CANCER CELLS AND OTHER LARGE CELLS FROM THE BLOOD. Cancer. 1964 May;17:637–642. doi: 10.1002/1097-0142(196405)17:5<637::aid-cncr2820170512>3.0.co;2-i. [DOI] [PubMed] [Google Scholar]
  18. Sarkar G., Sommer S. S. Shedding light on PCR contamination. Nature. 1990 Jan 4;343(6253):27–27. doi: 10.1038/343027a0. [DOI] [PubMed] [Google Scholar]
  19. Seddon J. M., Gragoudas E. S., Egan K. M., Glynn R. J., Howard S., Fante R. G., Albert D. M. Relative survival rates after alternative therapies for uveal melanoma. Ophthalmology. 1990 Jun;97(6):769–777. doi: 10.1016/s0161-6420(90)32512-5. [DOI] [PubMed] [Google Scholar]
  20. Shammas H. F., Blodi F. C. Prognostic factors in choroidal and ciliary body melanomas. Arch Ophthalmol. 1977 Jan;95(1):63–69. doi: 10.1001/archopht.1977.04450010065005. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Stanford G. B., Reese A. B. Malignant cells in the blood of eye patients. Trans Am Acad Ophthalmol Otolaryngol. 1971 Jan-Feb;75(1):102–109. [PubMed] [Google Scholar]
  23. TOBIN C. E., ZARIQUIEY M. O. Arteriovenous shunts in the human lung. Proc Soc Exp Biol Med. 1950 Dec;75(3):827–829. doi: 10.3181/00379727-75-18360. [DOI] [PubMed] [Google Scholar]
  24. Tobal K., Sherman L. S., Foss A. J., Lightman S. L. Detection of melanocytes from uveal melanoma in peripheral blood using the polymerase chain reaction. Invest Ophthalmol Vis Sci. 1993 Aug;34(9):2622–2625. [PubMed] [Google Scholar]
  25. Zimmerman L. E., McLean I. W. An evaluation of enucleation in the management of uveal melanomas. Am J Ophthalmol. 1979 Jun;87(6):741–760. doi: 10.1016/0002-9394(79)90348-9. [DOI] [PubMed] [Google Scholar]
  26. Zimmerman L. E., McLean I. W., Foster W. D. Does enucleation of the eye containing a malignant melanoma prevent or accelerate the dissemination of tumour cells. Br J Ophthalmol. 1978 Jun;62(6):420–425. doi: 10.1136/bjo.62.6.420. [DOI] [PMC free article] [PubMed] [Google Scholar]