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. 1999 Apr;79(11-12):1813–1820. doi: 10.1038/sj.bjc.6990289

Detection of cytokeratins 19/20 and guanylyl cyclase C in peripheral blood of colorectal cancer patients

S A Bustin 1, V G Gyselman 1, N S Williams 1, S Dorudi 1
PMCID: PMC2362804  PMID: 10206298

Abstract

The clinical significance of detecting supposed tumour cell-derived mRNA transcripts in blood using the polymerase chain reaction (PCR) remains unclear. We have used a fully quantitative 5′-nuclease RT-PCR assay to screen for the expression of cytokeratins (ck) 19 and 20 and guanylyl cyclase C (GCC) in the peripheral blood of 21 healthy controls and 27 colorectal cancer patients. Expression of cytokeratin 19 and 20 mRNA was detected in 30% and 100% of samples, respectively, taken from healthy volunteers. There was no apparent difference in ck19 and ck20 mRNA transcription levels between controls and patients, or between patients with different Dukes' stages. While GCC mRNA was detected in only 1/21 control samples, it was expressed in approximately 80% of patients, although again there was no correlation between GCC levels and disease stage. Transcription levels of all three markers varied considerably between samples, even between samples taken from the same person at different times. We conclude that neither ck19 nor ck20 are reliable markers for the detection of colon epithelial cells in peripheral blood and that an evaluation of the usefulness of GCC awaits further longitudinal studies. © 1999 Cancer Research Campaign

Keywords: solid tumours, micrometastases, cytokeratins, guanylyl cyclase C, RT-PCR, quantitation

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Selected References

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  1. Berg L. P., Wieland K., Millar D. S., Schlösser M., Wagner M., Kakkar V. V., Reiss J., Cooper D. N. Detection of a novel point mutation causing haemophilia A by PCR/direct sequencing of ectopically-transcribed factor VIII mRNA. Hum Genet. 1990 Oct;85(6):655–658. doi: 10.1007/BF00193593. [DOI] [PubMed] [Google Scholar]
  2. Boland C. R. Setting microsatellites free. Nat Med. 1996 Sep;2(9):972–974. doi: 10.1038/nm0996-972. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Bustin S. A., Dorudi S. Molecular assessment of tumour stage and disease recurrence using PCR-based assays. Mol Med Today. 1998 Sep;4(9):389–396. doi: 10.1016/s1357-4310(98)01324-0. [DOI] [PubMed] [Google Scholar]
  5. Carrithers S. L., Barber M. T., Biswas S., Parkinson S. J., Park P. K., Goldstein S. D., Waldman S. A. Guanylyl cyclase C is a selective marker for metastatic colorectal tumors in human extraintestinal tissues. Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14827–14832. doi: 10.1073/pnas.93.25.14827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Chelly J., Hugnot J. P., Concordet J. P., Kaplan J. C., Kahn A. Illegitimate (or ectopic) transcription proceeds through the usual promoters. Biochem Biophys Res Commun. 1991 Jul 31;178(2):553–557. doi: 10.1016/0006-291x(91)90143-u. [DOI] [PubMed] [Google Scholar]
  8. Cooper D. N., Berg L. P., Kakkar V. V., Reiss J. Ectopic (illegitimate) transcription: new possibilities for the analysis and diagnosis of human genetic disease. Ann Med. 1994 Feb;26(1):9–14. doi: 10.3109/07853899409147321. [DOI] [PubMed] [Google Scholar]
  9. Deans G. T., Parks T. G., Rowlands B. J., Spence R. A. Prognostic factors in colorectal cancer. Br J Surg. 1992 Jul;79(7):608–613. doi: 10.1002/bjs.1800790706. [DOI] [PubMed] [Google Scholar]
  10. Denis M. G., Lipart C., Leborgne J., LeHur P. A., Galmiche J. P., Denis M., Ruud E., Truchaud A., Lustenberger P. Detection of disseminated tumor cells in peripheral blood of colorectal cancer patients. Int J Cancer. 1997 Oct 21;74(5):540–544. doi: 10.1002/(sici)1097-0215(19971021)74:5<540::aid-ijc11>3.0.co;2-a. [DOI] [PubMed] [Google Scholar]
  11. Dingemans A. M., Brakenhoff R. H., Postmus P. E., Giaccone G. Detection of cytokeratin-19 transcripts by reverse transcriptase-polymerase chain reaction in lung cancer cell lines and blood of lung cancer patients. Lab Invest. 1997 Sep;77(3):213–220. [PubMed] [Google Scholar]
  12. Dorudi S., Kinrade E., Marshall N. C., Feakins R., Williams N. S., Bustin S. A. Genetic detection of lymph node micrometastases in patients with colorectal cancer. Br J Surg. 1998 Jan;85(1):98–100. doi: 10.1046/j.1365-2168.1998.02872.x. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Fukuhara T., Hooper W. C., Baylin S. B., Benson J., Pruckler J., Olson A. C., Evatt B. L., Vogler W. R. Use of the polymerase chain reaction to detect hypermethylation in the calcitonin gene. A new, sensitive approach to monitor tumor cells in acute myelogenous leukemia. Leuk Res. 1992 Oct;16(10):1031–1040. doi: 10.1016/0145-2126(92)90083-j. [DOI] [PubMed] [Google Scholar]
  15. Funaki N. O., Tanaka J., Itami A., Kasamatsu T., Ohshio G., Onodera H., Monden K., Okino T., Imamura M. Detection of colorectal carcinoma cells in circulating peripheral blood by reverse transcription-polymerase chain reaction targeting cytokeratin-20 mRNA. Life Sci. 1997;60(9):643–652. doi: 10.1016/s0024-3205(96)00700-x. [DOI] [PubMed] [Google Scholar]
  16. Gerhard M., Juhl H., Kalthoff H., Schreiber H. W., Wagener C., Neumaier M. Specific detection of carcinoembryonic antigen-expressing tumor cells in bone marrow aspirates by polymerase chain reaction. J Clin Oncol. 1994 Apr;12(4):725–729. doi: 10.1200/JCO.1994.12.4.725. [DOI] [PubMed] [Google Scholar]
  17. Glaves D., Huben R. P., Weiss L. Haematogenous dissemination of cells from human renal adenocarcinomas. Br J Cancer. 1988 Jan;57(1):32–35. doi: 10.1038/bjc.1988.4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gunn J., McCall J. L., Yun K., Wright P. A. Detection of micrometastases in colorectal cancer patients by K19 and K20 reverse-transcription polymerase chain reaction. Lab Invest. 1996 Oct;75(4):611–616. [PubMed] [Google Scholar]
  19. Hanekom G. S., Johnson C. A., Kidson S. H. An improved and combined reverse transcription-polymerase chain reaction assay for reliable detection of metastatic melanoma cells in peripheral blood. Melanoma Res. 1997 Apr;7(2):111–116. doi: 10.1097/00008390-199704000-00004. [DOI] [PubMed] [Google Scholar]
  20. Hayashi N., Ito I., Yanagisawa A., Kato Y., Nakamori S., Imaoka S., Watanabe H., Ogawa M., Nakamura Y. Genetic diagnosis of lymph-node metastasis in colorectal cancer. Lancet. 1995 May 20;345(8960):1257–1259. doi: 10.1016/s0140-6736(95)90922-2. [DOI] [PubMed] [Google Scholar]
  21. Holland P. M., Abramson R. D., Watson R., Gelfand D. H. Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7276–7280. doi: 10.1073/pnas.88.16.7276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Johnson P. W., Burchill S. A., Selby P. J. The molecular detection of circulating tumour cells. Br J Cancer. 1995 Aug;72(2):268–276. doi: 10.1038/bjc.1995.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jonas S., Windeatt S., O-Boateng A., Fordy C., Allen-Mersh T. G. Identification of carcinoembryonic antigen-producing cells circulating in the blood of patients with colorectal carcinoma by reverse transcriptase polymerase chain reaction. Gut. 1996 Nov;39(5):717–721. doi: 10.1136/gut.39.5.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Leygue E., Murphy L., Kuttenn F., Watson P. Triple primer polymerase chain reaction. A new way to quantify truncated mRNA expression. Am J Pathol. 1996 Apr;148(4):1097–1103. [PMC free article] [PubMed] [Google Scholar]
  25. Liefers G. J., Cleton-Jansen A. M., van de Velde C. J., Hermans J., van Krieken J. H., Cornelisse C. J., Tollenaar R. A. Micrometastases and survival in stage II colorectal cancer. N Engl J Med. 1998 Jul 23;339(4):223–228. doi: 10.1056/NEJM199807233390403. [DOI] [PubMed] [Google Scholar]
  26. Litle V. R., Warren R. S., Moore D., 2nd, Pallavicini M. G. Molecular cytogenetic analysis of cytokeratin 20-labeled cells in primary tumors and bone marrow aspirates from colorectal carcinoma patients. Cancer. 1997 May 1;79(9):1664–1670. [PubMed] [Google Scholar]
  27. Moll R., Löwe A., Laufer J., Franke W. W. Cytokeratin 20 in human carcinomas. A new histodiagnostic marker detected by monoclonal antibodies. Am J Pathol. 1992 Feb;140(2):427–447. [PMC free article] [PubMed] [Google Scholar]
  28. Moll R., Zimbelmann R., Goldschmidt M. D., Keith M., Laufer J., Kasper M., Koch P. J., Franke W. W. The human gene encoding cytokeratin 20 and its expression during fetal development and in gastrointestinal carcinomas. Differentiation. 1993 Jun;53(2):75–93. doi: 10.1111/j.1432-0436.1993.tb00648.x. [DOI] [PubMed] [Google Scholar]
  29. Mori M., Mimori K., Inoue H., Barnard G. F., Tsuji K., Nanbara S., Ueo H., Akiyoshi T. Detection of cancer micrometastases in lymph nodes by reverse transcriptase-polymerase chain reaction. Cancer Res. 1995 Aug 1;55(15):3417–3420. [PubMed] [Google Scholar]
  30. Nawroz H., Koch W., Anker P., Stroun M., Sidransky D. Microsatellite alterations in serum DNA of head and neck cancer patients. Nat Med. 1996 Sep;2(9):1035–1037. doi: 10.1038/nm0996-1035. [DOI] [PubMed] [Google Scholar]
  31. Quinlan R. A., Schiller D. L., Hatzfeld M., Achtstätter T., Moll R., Jorcano J. L., Magin T. M., Franke W. W. Patterns of expression and organization of cytokeratin intermediate filaments. Ann N Y Acad Sci. 1985;455:282–306. doi: 10.1111/j.1749-6632.1985.tb50418.x. [DOI] [PubMed] [Google Scholar]
  32. Sorg R., Enczmann J., Sorg U., Heermeier K., Schneider E. M., Wernet P. Rapid and sensitive mRNA phenotyping for interleukins (IL-1 to IL-6) and colony-stimulating factors (G-CSF, M-CSF, and GM-CSF) by reverse transcription and subsequent polymerase chain reaction. Exp Hematol. 1991 Oct;19(9):882–887. [PubMed] [Google Scholar]
  33. Tarin D., Price J. E., Kettlewell M. G., Souter R. G., Vass A. C., Crossley B. Mechanisms of human tumor metastasis studied in patients with peritoneovenous shunts. Cancer Res. 1984 Aug;44(8):3584–3592. [PubMed] [Google Scholar]
  34. Traweek S. T., Liu J., Battifora H. Keratin gene expression in non-epithelial tissues. Detection with polymerase chain reaction. Am J Pathol. 1993 Apr;142(4):1111–1118. [PMC free article] [PubMed] [Google Scholar]
  35. Waldman S. A., Cagir B., Rakinic J., Fry R. D., Goldstein S. D., Isenberg G., Barber M., Biswas S., Minimo C., Palazzo J. Use of guanylyl cyclase C for detecting micrometastases in lymph nodes of patients with colon cancer. Dis Colon Rectum. 1998 Mar;41(3):310–315. doi: 10.1007/BF02237484. [DOI] [PubMed] [Google Scholar]
  36. Wong L. S., Cantrill J. E., Odogwu S., Morris A. G., Fraser I. A. Detection of circulating tumour cells and nodal metastasis by reverse transcriptase-polymerase chain reaction technique. Br J Surg. 1997 Jun;84(6):834–839. [PubMed] [Google Scholar]
  37. Wyld D. K., Selby P., Perren T. J., Jonas S. K., Allen-Mersh T. G., Wheeldon J., Burchill S. A. Detection of colorectal cancer cells in peripheral blood by reverse-transcriptase polymerase chain reaction for cytokeratin 20. Int J Cancer. 1998 Jun 19;79(3):288–293. doi: 10.1002/(sici)1097-0215(19980619)79:3<288::aid-ijc14>3.0.co;2-4. [DOI] [PubMed] [Google Scholar]
  38. Zippelius A., Kufer P., Honold G., Köllermann M. W., Oberneder R., Schlimok G., Riethmüller G., Pantel K. Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow. J Clin Oncol. 1997 Jul;15(7):2701–2708. doi: 10.1200/JCO.1997.15.7.2701. [DOI] [PubMed] [Google Scholar]

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