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Multiplex RT-PCR-based detections of CEA, CK20 and EGFR in colorectal cancer patients
Aikaterini Tsouma, Chrysanthi Aggeli, Panagiotis Lembessis, George N Zografos, Dimitris P Korkolis, Dimitrios Pectasides, Maria Skondra, Nikolaos Pissimissis, Anastasia Tzonou, Michael Koutsilieris
Aikaterini Tsouma, Chrysanthi Aggeli, Maria Skondra, Nikolaos Pissimissis, Michael Koutsilieris, Department of Experimental Physiology, Medical School, National and Kapodistrian University of Athens, 11527, Athens, Greece
Chrysanthi Aggeli, George N Zografos, Third Department of Surgery, Athens General Hospital “G. Gennimatas”, 11527, Athens, Greece
Chrysanthi Aggeli, Dimitris P Korkolis, Department of Surgical Oncology, Hellenic Anticancer Institute, “Saint Savvas” Hospital, 11522, Athens, Greece
Panagiotis Lembessis, Endo/OncoResearch Medical Center, Platia Mavilis, Ampelokipi, 11521, Athens, Greece
Dimitrios Pectasides, Maria Skondra, Second Department of Internal Medicine, Propaedeutic Oncology Section, University of Athens, Attikon University Hospital, 12462, Athens, Greece
Anastasia Tzonou, Department of Hygiene, Epidemiology and Medical Statistics, Medical School, National and Kapodistrian University of Athens, 11527, Athens, Greece
Author contributions: Tsouma A, Aggeli C, Lembessis P and Koutsilieris M developed the concept of the multiplex PCR-based technique, performed data analysis and wrote the present manuscript; Aggeli C, Zografos GN, Korkolis DP, Pectasides D, Skondra M and Pissimissis N collected blood samples and clinical data, evaluated the clinical data, performed data analysis and participated in the writing of the manuscript; Tzonou A performed statistical analysis and participated in the writing of the manuscript.
Supported by The Ministry of Development of the Greek Government (GGET-AKMON)
Correspondence to: Michael Koutsilieris, MD, PhD, Professor and Chairman, Department of Experimental Physiology, Medical School, National and Kapodistrian University of Athens, Micras Asias 75, Goudi-Athens, 11527, Athens, Greece. mkoutsil@med.uoa.gr
Telephone: +30-210-7462597 Fax: +30-210-7462571
Received: May 17, 2010
Revised: July 12, 2010
Accepted: July 19, 2010
Published online: December 21, 2010
AIM: To develop a multiplex reverse transcription polymerase chain reaction (RT-PCR) method detecting circulating tumor cells in the peripheral blood of colorectal cancer (CRC) patients.
METHODS: Peripheral blood samples were collected from 88 CRC patients and 40 healthy individuals from the blood donors’ clinic and subsequently analyzed by multiplex RT-RCR for the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and epidermal growth factor receptor (EGFR) mRNA. The analysis involved determining the detection rates of CEA, CK20 and EGFR transcripts vs disease stage and overall survival. Median follow-up period was 19 mo (range 8-28 mo).
RESULTS: Rates of CEA, CK20 and EGFR detection in CRC patients were 95.5%, 78.4% and 19.3%, respectively. CEA transcripts were detected in 3 healthy volunteer samples (7.5%), whereas all control samples were tested negative for CK20 and EGFR transcripts. The increasing number of positive detections for CEA, CK20 and EGFR transcripts in each blood sample was positively correlated with Astler-Coller disease stage (P < 0.001) and preoperative serum levels of CEA (P = 0.029) in CRC patients. Data analysis using Kaplan-Meier estimator documented significant differences in the overall survival of the different CRC patient groups as formed according to the increasing number of positivity for CEA, CK20 and EGFR transcripts.
CONCLUSION: These data suggest that multiplex RT-PCR assay can provide useful information concerning disease stage and overall survival of CRC patients.
