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Detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR for CEA and CK20 mRNAS

Oberg, ANV; Lindmark, Gudrun LU ; Israelsson, ACE; Hammarstrom, SG and Hammarstrom, MLCK (2004) In International Journal of Cancer 111(1). p.101-110
Abstract
The purpose of our study was to develop specific, sensitive, objective assays for early detection of disseminated tumour cells in patients with colorectal cancer (CRC). Carcinoembryonic antigen (CEA) and cytokeratin 20 (CK20) were chosen as markers because they are selectively expressed in epithelial cells with maintained expression in CRC. Real-time quantitative RT-PCR assays with RNA copy standards were constructed. Regional lymph nodes were collected from patients with CRC (n = 5 1) and benign intestinal disease (n = 10). Results were compared to routine histopathology and anti-CEA immunohistochemistry. Lymph node levels of CEA and CK20 mRNA correlated strongly (p < 0.0001, r = 0.8). Lymph nodes from non-CRC patients had <0.01 CEA... (More)
The purpose of our study was to develop specific, sensitive, objective assays for early detection of disseminated tumour cells in patients with colorectal cancer (CRC). Carcinoembryonic antigen (CEA) and cytokeratin 20 (CK20) were chosen as markers because they are selectively expressed in epithelial cells with maintained expression in CRC. Real-time quantitative RT-PCR assays with RNA copy standards were constructed. Regional lymph nodes were collected from patients with CRC (n = 5 1) and benign intestinal disease (n = 10). Results were compared to routine histopathology and anti-CEA immunohistochemistry. Lymph node levels of CEA and CK20 mRNA correlated strongly (p < 0.0001, r = 0.8). Lymph nodes from non-CRC patients had <0.01 CEA and <0.001 CK20 mRNA copies/18S rRNA unit. Lymph nodes from 3/6 Dukes' A, 17/26 Dukes' B, 10/10 Dukes' C and 7/9 Dukes' D patients had CEA mRNA levels above cut-off. Corresponding figures for CK20 mRNA were 3/6, 10/26, 9/10 and 5/9, respectively. CEA mRNA levels varied from 0.001 to 100 copies/18S rRNA unit in Dukes' A and B, and 50% of the Dukes' B patients had CEA mRNA levels within the range of Dukes' C patients. Three Dukes' B patients have died from CRC or developed distant metastases. All 3 had high CEA and CK20 mRNA levels. Determination of mRNA was superior to immunohistochemistry in showing CEA expression in lymph nodes. The present qRT-PCR assay for CEA mRNA seems to be a superior tool to identify individuals with disseminated tumour cells. Future extended studies will establish the clinically most relevant cut-off level. (C) 2004 Wiley-Liss, Inc. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
minimal residual, cancer, lymph node metastases, real-time quantitative RT-PCR, cytokeratin 20, colorectal neoplasm, carcinoembryonic antigen
in
International Journal of Cancer
volume
111
issue
1
pages
101 - 110
publisher
John Wiley & Sons
external identifiers
  • pmid:15185350
  • wos:000222445500015
  • scopus:3042775478
ISSN
0020-7136
DOI
10.1002/ijc.20231
language
English
LU publication?
yes
id
a6f581eb-5583-4913-a666-06a86bb6ae8e (old id 273530)
date added to LUP
2007-10-29 10:17:45
date last changed
2017-10-01 03:43:51
@article{a6f581eb-5583-4913-a666-06a86bb6ae8e,
  abstract     = {The purpose of our study was to develop specific, sensitive, objective assays for early detection of disseminated tumour cells in patients with colorectal cancer (CRC). Carcinoembryonic antigen (CEA) and cytokeratin 20 (CK20) were chosen as markers because they are selectively expressed in epithelial cells with maintained expression in CRC. Real-time quantitative RT-PCR assays with RNA copy standards were constructed. Regional lymph nodes were collected from patients with CRC (n = 5 1) and benign intestinal disease (n = 10). Results were compared to routine histopathology and anti-CEA immunohistochemistry. Lymph node levels of CEA and CK20 mRNA correlated strongly (p &lt; 0.0001, r = 0.8). Lymph nodes from non-CRC patients had &lt;0.01 CEA and &lt;0.001 CK20 mRNA copies/18S rRNA unit. Lymph nodes from 3/6 Dukes' A, 17/26 Dukes' B, 10/10 Dukes' C and 7/9 Dukes' D patients had CEA mRNA levels above cut-off. Corresponding figures for CK20 mRNA were 3/6, 10/26, 9/10 and 5/9, respectively. CEA mRNA levels varied from 0.001 to 100 copies/18S rRNA unit in Dukes' A and B, and 50% of the Dukes' B patients had CEA mRNA levels within the range of Dukes' C patients. Three Dukes' B patients have died from CRC or developed distant metastases. All 3 had high CEA and CK20 mRNA levels. Determination of mRNA was superior to immunohistochemistry in showing CEA expression in lymph nodes. The present qRT-PCR assay for CEA mRNA seems to be a superior tool to identify individuals with disseminated tumour cells. Future extended studies will establish the clinically most relevant cut-off level. (C) 2004 Wiley-Liss, Inc.},
  author       = {Oberg, ANV and Lindmark, Gudrun and Israelsson, ACE and Hammarstrom, SG and Hammarstrom, MLCK},
  issn         = {0020-7136},
  keyword      = {minimal residual,cancer,lymph node metastases,real-time quantitative RT-PCR,cytokeratin 20,colorectal neoplasm,carcinoembryonic antigen},
  language     = {eng},
  number       = {1},
  pages        = {101--110},
  publisher    = {John Wiley & Sons},
  series       = {International Journal of Cancer},
  title        = {Detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR for CEA and CK20 mRNAS},
  url          = {http://dx.doi.org/10.1002/ijc.20231},
  volume       = {111},
  year         = {2004},
}