title: Multiplex RT-PCR-based detections of CEA, CK20 and EGFR in colorectal cancer patients.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  These data suggest that multiplex RT-PCR assay can provide useful information concerning disease stage and overall survival of CRC patients.
authors.affiliation: Department of Experimental Physiology, Medical School, National and Kapodistrian University of Athens, Micras Asias 75, Goudi-Athens, 11527 Athens, Greece.
authors.forename: Aikaterini
authors.initials: A
authors.lastname: Tsouma
authors.forename: Chrysanthi
authors.initials: C
authors.lastname: Aggeli
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Lembessis
authors.forename: George-N
authors.initials: GN
authors.lastname: Zografos
authors.forename: Dimitris-P
authors.initials: DP
authors.lastname: Korkolis
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Pectasides
authors.forename: Maria
authors.initials: M
authors.lastname: Skondra
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Pissimissis
authors.forename: Anastasia
authors.initials: A
authors.lastname: Tzonou
authors.forename: Michael
authors.initials: M
authors.lastname: Koutsilieris
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002272:Carcinoembryonic Antigen / Q000097:blood* / Q000235:genetics; D015179:Colorectal Neoplasms / Q000097:blood* / Q000235:genetics / Q000473:pathology; D066246:ErbB Receptors / Q000097:blood* / Q000235:genetics; D005260:Female; D006801:Humans; D053208:Kaplan-Meier Estimate; D053554:Keratin-20 / Q000097:blood* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D002272:Carcinoembryonic Antigen; D053554:Keratin-20; D066246:ErbB Receptors