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01-09-2007 | Original Article

Detection of disseminated tumor cells in the lymph nodes of colorectal cancer patients using a real-time polymerase chain reaction assay

Authors: Erkki Lotspeich, Markus Schoene, Heinz Gerngroß, Roland Schmidt, Reinhard Steinmann, Marco Ramadani, Susanne Gansauge

Published in: Langenbeck's Archives of Surgery | Issue 5/2007

Abstract

Introduction

Postoperative treatment for colorectal cancer depends on tumor stage as defined by the International Union Against Cancer (UICC). Adjuvant chemotherapy is not recommended in patients without lymph node involvement (UICC stages I and II). As many as 20–30% of these patients, however, will develop recurrence.

Aims and objectives

We conducted this study to determine the presence of disseminated tumor cells in the lymph nodes by quantitative real-time polymerase chain reaction (QRT-PCR) for cytokeratin 20 (CK20) in an attempt to provide supplementary information compared to histopathological findings.

Materials and methods

Using a standard QRT-PCR assay, we examined primary tumors and 391 lymph nodes from 31 patients with completely resected colorectal cancer.

Results

Of the 31 primary tumors, 29 were positive for CK20 by QRT-PCR.

Discussion

An examination of the lymph nodes from the 29 patients with CK20-positive primary tumors revealed that 35 (92.1% sensitivity) of the 38 histopathologically positive lymph nodes and 54 (16.7%) of the 324 histopathologically negative lymph nodes were positive by molecular analysis. CK20 expression was detected in 10 (100%) of 10 patients with a histopathologically positive lymph node status (pN1). In 9 (47.4%) of 19 patients with negative histopathological results (pN0), we detected a CK20 mRNA signal in at least one lymph node. Whereas eight patients with histopathologically negative lymph nodes could be upstaged on the basis of the molecular findings, no patient would be downstaged.

Conclusion

Our results suggest that QRT-PCR for CK20 is a useful tool for the quantitative detection of micrometastases in the regional lymph nodes. We introduce a standardized procedure that integrates a molecular diagnostic technique in the clinical staging.

Parkin DM, Bray FI, Devesa SS (2001) Cancer burden in the year 2000. The global picture. Eur J Cancer 37:4–66CrossRef

Herold G (2004) Innere medizin 2004. Private, Cologne, pp 413–418

Gastinger I, Marusch F (2001) Evidence-based surgery in colon carcinoma. Zentralbl Chir 126:283–288PubMedCrossRef

Wiggers T, Jeekel J, Arends JW, Brinkhorst AP, Kluck HM, Luyk CI, Munting JD, Povel JA, Rutten AP, Volovics A (1988) No-touch isolation technique in colon cancer: a controlled prospective trial. Br J Surg 75:409–415PubMedCrossRef

Lehnert T, Herfarth Ch (1996) Principles and value of lymph node excision in colorectal carcinoma. Chirurg 67:889–899PubMedCrossRef

Ratto C, Sofo L, Ippoliti M, Merico M, Doglietto GB, Crucitti F (1998) Prognostic factors in colorectal cancer. Literature review for clinical application. Dis Colon Rectum 41:1033–1049PubMedCrossRef

Pantel K, von Knebel Doebritz M, Izbicki JR, Riethmueller G (1997) Disseminated tumor cells: diagnosis, prognostic relevance, phenotyping and therapeutic strategies. Chirurg 68:1241–1250PubMedCrossRef

Midgley R, Kerr D (1999) Colorectal cancer. Lancet 353:391–399PubMedCrossRef

Yarbro JW, Page DL, Fielding LP, Partridge EE, Murphy GP (1999) American Joint Committee on Cancer prognostic factors consensus conference. Cancer 86:2436–2446PubMedCrossRef

10.

Takahashi T, Kato T, Kodaira S, Koyama Y, Sakabe T, Tominaga T, Hamano K, Yasutomi M, Ogawa N (1996) Prognostic factors of colorectal cancer. Results of multivariate analysis of curative resection cases with or without adjuvant chemotherapy. Am J Clin Oncol 19:408–415PubMedCrossRef

11.

Futumura M, Takagi Y, Koumura H, Kida H, Tanemura H, Shimokawa K, Saji S (1998) Spread of colorectal cancer micrometastases in regional lymph nodes by reverse transcriptase-polymerase chain reactions for carcinoembryonic antigen and cytokeratin 20. J Surg Oncol 68:34–40CrossRef

12.

Miyake Y, Yamamoto H, Fujiwara Y, Ohue M, Sugita Y, Tomita N, Sekimoto M, Matsuura N, Shiozaki H, Monden M (2001) Extensive micrometastases to lymph nodes as a marker for rapid recurrence of colorectal cancer: a study of lymphatic mapping. Clin Cancer Res 7:1350–1357PubMed

13.

Pantel K, von Knebel Doeberitz M (2000) Detection and clinical relevance of micrometastatic cancer cells. Curr Opin Oncol 12:95–101PubMedCrossRef

14.

Rosenberg R, Hoos A, Mueller J, Baier P, Stricker D, Werner M, Nekarda H, Siewert JRl (2002) Prognostic significance of cytokeratin-20 reverse transcriptase polymerase chain reaction in lymph nodes of node-negative colorectal cancer patients. J Clin Oncol 20:1049–1055PubMedCrossRef

15.

Tsavalles G, Patel H, Allen-Mersh TG (2001) Detection and clinical significance of occult tumor cells in colorectal cancer. Br J Surg 88:1307–1320CrossRef

16.

Yamamoto N, Kato Y, Yanagisawa A, Ohta H, Takahashi T, Kitagawa T (1997) Predictive value of genetic diagnosis for cancer micrometastasis: histologic and experimental appraisal. Cancer 80:1393–1398PubMedCrossRef

17.

Moll R, Zimbelmann R, Goldschmidt MD, Keith M, Laufer J, Kasper M, Koch PJ, Franke WW (1993) The human gene encoding cytokeratin 20 and its expression during fetal development and in gastrointestinal carcinomas. Differentiation 53:75–93PubMedCrossRef

18.

Nagle RB (1988) Intermediate filaments: a review of the basic biology. Am J Surg Pathol 12:4–16PubMed

19.

Nagle RB (1989) Intermediate filaments. Efficacy in surgical pathologic diagnosis. Am J Clin Pathol 91:14–18

20.

Osborn M, Weber K (1983) Tumor diagnosis by intermediate filament typing: a novel tool for surgical pathology. Lab Invest 48:372–394PubMed

21.

Bustin SA, Gyselmann VG, Williams NS, Dorudi S (1999) Detection of cytokeratins 19/20 and guanyl cyclase C in peripheral blood of colorectal cancer patients. Br J Cancer 79:1813–1820PubMedCrossRef

22.

Funaki NO, Tamaka J, Ohshio G, Onodera H, Maetani S, Imamura M (1998) Cytokeratin 20 mRNA in peripheral venous blood of colorectal carcinoma patients. Br J Cancer 77:1327–1332PubMed

23.

Wharton RQ, Jonas SK, Glover C, Khan ZA, Klokouzas A, Quinn H, Henry M, Allen-Mersh TG (1999) Increased detection of circulating tumor cells in the blood of colorectal carcinoma patients using two reverse transcription-PCR assays and multiple blood samples. Clin Cancer Res 5:4158–4163PubMed

24.

Soeth E, Vogel I, Roder C, Juhl H, Marxsen J, Kruger U, Henne-Bruns D, Kremer B, Kalthoff H (1997) Comparative analysis of bone marrow and venous blood isolates from gastrointestinal cancer patients for the detection of disseminated tumor cells using reverse transcription PCR. Cancer Res 57:3106–3110PubMed

25.

Lassmann S, Bauer M, Rosenberg R, Nekarda H, Soong R, Ruger R, Hofler H, Werner M (2004) Identification of occult tumor cells in node negative lymph nodes of colorectal cancer patients by cytokeratin 20 gene and protein expression. Int J Colorectal Dis 19:87–94PubMedCrossRef

26.

Merrie AE, van Rij AM, Dennett ER, Phillips LV, Yun K, McCall JL (2003) Prognostic significance of occult metastases in colon cancer. Dis Colon Rectum 46:221–231PubMedCrossRef

27.

Jung R, Petersen K, Kruger W, Wolf M, Wagener C, Zander A, Neumaier M (1999) Detection of micrometastasis by cytokeratin 20 RT-PCR is limited due to stable background transcription in granulocytes. Br J Cancer 81:870–873PubMedCrossRef

28.

Soong R, Beyser K, Basten O, Kalbe A, Rueschoff J, Tabiti K (2001) Quantitative reverse transcription-polymerase chain reaction detection of cytokeratin 20 in noncolorectal lymph nodes. Clin Cancer Res 7:3423–3429PubMed

29.

Vlems FA, Diepstra JH, Cornelissen IM, Ruers TJ, Ligtenberg MJ, Punt CJ, van Krieken JH, Wobbes T, van Muijen GN (2002) Limitations of cytokeratin 20 RT-PCR to detect disseminated tumour cells in blood and bone marrow of patients with colorectal cancer: expression in controls and downregulation in tumour tissue. Mol Pathol 55:156–163PubMedCrossRef

30.

Weitz J, Kienle P, Magener A, Koch M, Schrodel A, Willeke F, Autschbach F, Lacroix J, Lehnert T, Herfarth C, von Knebel Doeberitz M (1999) Detection of disseminated colorectal cancer cells in lymph nodes, blood and bone marrow. Clin Cancer Res 5:1830–1836PubMed

31.

Liefers GJ, Cleton-Jansen AM, van de Velde CJ, Hermans J, van Krieken JH, Cornelisse CJ, Tollenaar RA (1998) Micrometastases and survival in stage II colorectal cancer. N Engl J Med 339:223–228PubMedCrossRef

32.

Lassmann S, Bauer M, Soong R, Schreglmann J, Tabiti K, Nahrig J, Ruger R, Hofler H, Werner M (2002) Quantification of CK20 gene and protein expression in colorectal cancer by RT-PCR and immunohistochemistry reveals inter- and intratumour heterogeneity. J Pathol 198:198–206PubMedCrossRef

Title: Detection of disseminated tumor cells in the lymph nodes of colorectal cancer patients using a real-time polymerase chain reaction assay
Authors: Erkki Lotspeich
Markus Schoene
Heinz Gerngroß
Roland Schmidt
Reinhard Steinmann
Marco Ramadani
Susanne Gansauge
Publication date: 01-09-2007
Publisher: Springer-Verlag
Published in: Langenbeck's Archives of Surgery / Issue 5/2007
Print ISSN: 1435-2443
Electronic ISSN: 1435-2451
DOI: https://doi.org/10.1007/s00423-007-0184-3

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Detection of disseminated tumor cells in the lymph nodes of colorectal cancer patients using a real-time polymerase chain reaction assay

Abstract

Introduction

Aims and objectives

Materials and methods

Results

Discussion

Conclusion

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Introduction

Aims and objectives

Materials and methods

Results

Discussion

Conclusion

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