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CLC number: R735.3; R73-37

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Received: 2006-01-09

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Journal of Zhejiang University SCIENCE B 2006 Vol.7 No.6 P.445-451


Quantitative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood of colorectal cancer patients

Author(s):  XU Dong, LI Xu-fen, ZHENG Shu, JIANG Wen-zhi

Affiliation(s):  Cancer Institute, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China

Corresponding email(s):   honokiol@gmail.com, zhengshu@zju.edu.cn

Key Words:  Colorectal carcinoma, Real-time RT-PCR, CEA mRNA, CK20 mRNA, CK19 mRNA

XU Dong, LI Xu-fen, ZHENG Shu, JIANG Wen-zhi. Quantitative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood of colorectal cancer patients[J]. Journal of Zhejiang University Science B, 2006, 7(6): 445-451.

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author="XU Dong, LI Xu-fen, ZHENG Shu, JIANG Wen-zhi",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Quantitative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood of colorectal cancer patients
%A XU Dong
%A LI Xu-fen
%A JIANG Wen-zhi
%J Journal of Zhejiang University SCIENCE B
%V 7
%N 6
%P 445-451
%@ 1673-1581
%D 2006
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2006.B0445

T1 - Quantitative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood of colorectal cancer patients
A1 - XU Dong
A1 - LI Xu-fen
A1 - ZHENG Shu
A1 - JIANG Wen-zhi
J0 - Journal of Zhejiang University Science B
VL - 7
IS - 6
SP - 445
EP - 451
%@ 1673-1581
Y1 - 2006
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2006.B0445

This study is aimed at establishing a sensitive approach to detect disseminated tumor cells in peripheral blood and evaluate its clinical significance. A total of 198 blood samples including 168 from colorectal carcinoma (CRC) patients and 30 from healthy volunteers were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and cytokeratin 19 (CK19) mRNA. CEA mRNA was detected in 35.8% of patients and 3.3% of controls, CK20 mRNA in 28.3% of patients and 6.7% of controls, and CK19 mRNA in 41.9% of patients and 3.3% of controls. CEA and CK20 mRNA positive ratio increased with the advancing Dukes stages, but there was no significant difference in positive ratio between any two stages (P>0.05). Also, relatively high positive ratio of CEA, CK20 and CK19 mRNA expression was observed in some CRC patients with earlier Dukes stages. A higher positive ratio was obtained when two or three detection markers were combined compared to a single marker. Our study indicates that quantitative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood is a valuable tool for monitoring early stage dissemination of CRC cells in blood circulation.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1] Chen, Y.H., Gao, W., Zhou, T., Zhao, W., Zhao, H., Liu, D., Felber, E., 1999. Detection of bone marrow micrometastasis. Hybridoma, 18(5):465-466.

[2] Dandachi, N., Balic, M., Stanzer, S., Halm, M., Resel, M., Hinterleitner, T.A., Samonigg, H., Bauernhofer, T., 2005. Critical evaluation of real-time reverse transcriptase-polymerase chain reaction for the quantitative detection of cytokeratin 20 mRNA in colorectal cancer patients. Mol. Diagn., 7(5):631-637.

[3] Deans, G.T., Parks, T.G., Rowlands, B.J., Spence, R.A., 1992. Prognostic factors in colorectal cancer. Br. J. Surg., 79(7):608-613.

[4] Fiorella, G., Judith, K., Simona, A., Maria, D.C., Antonella, S., Roberta, D.A., Maria, R.A., Maurizio, C., Franco, G., Fabio, C., et al., 2001. Detection of blood-borne cells in colorectal cancer patients by nested reverse transcription-polymerase chain reaction for carcinoembryonic antigen messenger RNA: longitudinal analyses and demonstration of its potential importance as an adjunct to multiple serum markers. Cancer Res., 61(6):2523-2532.

[5] Funaki, N.O., Tanaka, J., Itami, A., Kasamatsu, T., Ohshio, G., Onodera, H., Monden, K., Okino, T., Imamura, M., 1997. Detection of colorectal carcinoma cells in circulating peripheral blood by reverse transcription-polymerase chain reaction targeting cytokeratin-20 mRNA. Life Sci., 60(9):643-652.

[6] Giribaldi, G., Procida, S., Ulliers, D., Mannu, F., Volpatto, R., Mandili, G., Fanchini, L., Bertetto, O., Fronda, G., Simula, L., et al., 2006. Specific detection of cytokeratin 20-positive cells in blood of colorectal and breast cancer patients by a high sensitivity real-time reverse transcriptase-polymerase chain reaction method. Mol. Diagn., 8(1):105-112.

[7] Gradilone, A., Gazzaniga, P., Silvestri, I., Gandini, O., Trasatti, L., Lauro, S., Frati, L., Agliano, A.M., 2003. Detection of CK19, CK20 and EGFR mRNAs in peripheral blood of carcinoma patients: correlation with clinical stage of disease. Oncol. Rep., 10(1):217-222.

[8] Hardingham, J.E., Hewett, P.J., Sage, R.E., Finch, J.L., Nuttall, J.D., Kotasek, D., Dobrovic, A., 2000. Molecular detection of blood-borne epithelial cells in colorectal cancer patients and in patients with benign bowel disease. Int. J. Cancer, 89(1):8-13.

[9] Holdenrieder, S., Holubec, L.Jr, Topolcan, O., Finek, J., Stieber, P., 2005. Circulating nucleosomes and cytokeratin 19-fragments in patients with colorectal cancer during chemotherapy. Anticancer Res., 25(3A):1795-1801.

[10] Iinuma, H., Okinaga, K., Egami, H., Mimori, K., Hayashi, N., Nishida, K., Adachi, M., Mori, M., Sasako, M., 2006. Usefulness and clinical significance of quantitative real-time RT-PCR to detect isolated tumor cells in the peripheral blood and tumor drainage blood of patients with colorectal cancer. Int. J. Oncol., 28(2):297-306.

[11] Ikeda, S., Fujimori, M., Shibata, S., Okajima, M., Ishizaki, Y., Kurihara, T., Miyata, Y., Iseki, M., Shimizu, Y., Tokumoto, N., et al., 2006. Combined immunohistochemistry of beta-catenin, cytokeratin 7, and cytokeratin 20 is useful in discriminating primary lung adenocarcinomas from metastatic colorectal cancer. BMC Cancer, 6(1):31.

[12] Ikeguchi, M., Ohro, S., Maeda, Y., Fukuda, K., Yamaguchi, K., Shirai, H., Kondo, A., Tsujitani, S., Kaibara, N., 2003. Detection of cancer cells in the peripheral blood of gastric cancer patients. Int. J. Mol. Med., 11(2):217-221.

[13] Ishida, H., Miwa, H., Tatsuta, M., Masutani, S., Imamura, H., Shimizu, J., Ezumi, K., Kato, H., Kawasaki, T., Furukawa, H., et al., 2004. Ki-67 and CEA expression as prognostic markers in Dukes’ C colorectal cancer. Cancer Lett., 207(1):109-115.

[14] Jung, R., Petersen, K., Kruger, W., 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(5):870-873.

[15] Kijima, M., Togo, S., Ichikawa, Y., Miura, M., Yamagishi, S., Matsuo, K., Tanaka, K., Masui, H., Ishikawa, T., Ike, H., et al., 2005. Clinical significance of serum CEA protein and CEA mRNA after resection of colorectal liver metastases. Anticancer Res., 25(2B):1327-1332.

[16] Ko, Y., Grunewald, E., Totzke, G., Klinz, M., Fronhoffs, S., Gouni-Berthold, I., Sachinidis, A., Vetter, H., 2000. High percentage of false-positive results of cytokeratin 19 RT-PCR in blood: a model for the analysis of illegitimate gene expression. Oncology, 59(1):81-88.

[17] Leather, A.J., Gallegos, N.C., Kocjan, G., Savage, F., Smales, C.S., Hu, W., Boulos, P.B., Northover, J.M., Phillips, R.K., 1993. Detection and enumeration of circulating tumour cells in colorectal cancer. Br. J. Surg., 80(6):777-780.

[18] Miura, M., Ichikawa, Y., Tanaka, K., Kamiyama, M., Hamaguchi, Y., Ishikawa, T., Yamaguchi, S., Togo, S., Ike, H., Ooki, S., et al., 2003. Real-time PCR (TaqMan PCR) quantification of carcinoembryonic antigen (CEA) mRNA in the peripheral blood of colorectal cancer patients. Anticancer Res., 23(2B):1271-1276.

[19] Molnar, B., Sipos, F., Galamb, O., Tulassay, Z., 2003. Molecular detection of circulating cancer cells. Role in diagnosis, prognosis and follow-up of colon cancer patients. Dig. Dis., 21(4):320-325.

[20] Öberg, A.N., Lindmark, G.E., Israelsson, A.C., Hammarstrom, S.G., Hammarstrom, M.L., 2004. Detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR for CEA and CK20 mRNAS. Int. J. Cancer, 111(1):101-110.

[21] Oki, E., Maehara, Y., Tokunaga, E., Shibahara, K., Hasuda, S., Kakeji, Y., Sugimachi, K., 2002. Detection of disseminated cancer cells in bone marrow of gastric cancer using real time quantitative reverse transcriptase polymerase chain reaction. Cancer Lett., 188(1-2):191-198.

[22] Piva, M.G., Navaglia, F., Basso, D., Fogar, P., Roveroni, G., Gallo, N., Zambon, C.F., Pedrazzoli, S., Plebani, M., 2000. CEA mRNA identification in peripheral blood is feasible for colorectal, but not for gastric or pancreatic cancer staging. Oncology, 59(4):323-328.

[23] Rosenberg, R., Hoos, A., Mueller, J., Nekarda, H., 2000. Impact of cytokeratin-20 and carcinoembryonic antigen mRNA detection by RT-PCR in regional lymph nodes of patients with colorectal cancer. Br. J. Cancer, 83(10):1323-1329.

[24] Schuster, R., Max, N., Mann, B., Heufelder, K., Thilo, F., Grone, J., Rokos, F., Buhr, H.J., Thiel, E., Keilholz, U., 2004. Quantitative real-time RT-PCR for detection of disseminated tumor cells in peripheral blood of patients with colorectal cancer using different mRNA markers. Int. J. Cancer, 108(2):219-227.

[25] Stathopoulou, A., Gizi, A., Perraki, M., Apostolaki, S., Malamos, N., Mavroudis, D., Georgoulias, V., Lianidou, E.S., 2003. Real-time quantification of CK-19 mRNA-positive cells in peripheral blood of breast cancer patients using the lightcycler system. Clin. Cancer Res., 9(14):5145-5151.

[26] Vlems, F.A., Diepstra, J.H.S., Cornelissen, I.M.H.A., Ruers, T.J.M.M., Ligtenberg, J.L., Punt, C.J.A., Krieken, J.H.J.M., Wobbes, T., Muijen, G.N.P., 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. Molecular Pathology, 55(3):156-163.

[27] Wong, I.H., Yeo, W., Chan, A.T., Johnson, P.J., 2001. Quantitative relationship of the circulating tumor burden assessed by reverse transcription-polymerase chain reaction for cytokeratin 19 mRNA in peripheral blood of colorectal cancer patients with Dukes’ stage, serum carcinoembryonic antigen level and tumor progression. Cancer Lett., 162(1):65-73.

[28] Zhang, X.W., Fan, P., Yang, H.Y., Yang, L., Chen, G.Y., 2003. Significance of detecting disseminated tumor cells in peripheral blood of gastric and colorectal cancer patients. Zhonghua Zhong Liu Za Zhi, 25(1):66-69 (in Chinese).

[29] Zippelius, A., Kufer, P., Honold, G., Kollermann, M.W., Oberneder, R., Schlimok, G., Riethmuller, G., Pantel, K., 1997. Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow. J. Clin. Oncol., 15(7):2701-2708.

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