title: Disseminated single tumor cells as detected by real-time quantitative polymerase chain reaction represent a prognostic factor in patients undergoing surgery for colorectal cancer.
abstract: OBJECTIVE
  To evaluate the clinical relevance of real-time quantitative polymerase chain reaction (qPCR) detection of CEA and CK20 transcripts, as potentially related to tumor cell dissemination, in blood and peritoneal lavage from patients undergoing surgery for colorectal cancer.
  
  
  BACKGROUND
  Dissemination of single colorectal cancer cells in the peritoneal cavity, as well as in tumor drainage and peripheral blood vessels, might play a role in the metastasis process, thus affecting the clinical course. However, this phenomenon needs further elucidation.
  
  
  METHODS
  In a prospective study the authors evaluated the potential of qPCR in the detection of CEA and/or CK20 transcripts in the peritoneal lavage fluid and in the peripheral and mesenteric venous blood of 39 patients undergoing curative resection for colorectal cancer. Peritoneal lavage and peripheral blood was sampled before and after tumor resection; mesenteric venous blood was sampled from the major tumor-draining vein immediately before clamping. After RNA extraction and reverse transcription, qPCR was performed using specific cDNA primers and probes for CEA and CK20. The dichotomous results from the qPCR were used as a predictor along with other covariates in Cox proportional hazard regression models of long-term outcome (disease-free survival and overall survival).
  
  
  RESULTS
  Of 39 patients, 11 were positive. The median follow-up at analysis was 31 months for all patients. The dichotomous qPCR covariate was significant, with P =.001 and.0035 for disease-free survival and overall survival, respectively, in the proportional hazard regression models with only qPCR. In seven patients, disseminated colorectal cancer cells were found in the peritoneal lavage fluid but not in blood specimens; five of these patients (71%) had recurrence.
  
  
  CONCLUSIONS
  These data suggest that detection of mRNA coding for CEA and/or CK20 using qPCR has potential clinical utility as a prognostic marker and should be evaluated in larger clinical studies. Identification of patients at high risk for metastatic disease after curative resection of colorectal cancer might be improved by analyzing peritoneal lavage specimens in addition to blood samples. This is based on the observation that in more than half of qPCR-positive patients, disseminated colorectal cancer cells were detected in peritoneal lavage specimens but not in blood samples, and that 71% of them had recurrence.
authors.affiliation: Surgical Research Unit, Deparment of Surgery, University of Basel, Switzerland.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Guller
authors.forename: Paul
authors.initials: P
authors.lastname: Zajac
authors.forename: Annelies
authors.initials: A
authors.lastname: Schnider
authors.forename: Beatrix
authors.initials: B
authors.lastname: Bösch
authors.forename: Stefan
authors.initials: S
authors.lastname: Vorburger
authors.forename: Markus
authors.initials: M
authors.lastname: Zuber
authors.forename: Giulio Cesare
authors.initials: GC
authors.lastname: Spagnoli
authors.forename: Daniel
authors.initials: D
authors.lastname: Oertli
authors.forename: Robert
authors.initials: R
authors.lastname: Maurer
authors.forename: Urs
authors.initials: U
authors.lastname: Metzger
authors.forename: Felix
authors.initials: F
authors.lastname: Harder
authors.forename: Michael
authors.initials: M
authors.lastname: Heberer
authors.forename: Walter Richard
authors.initials: WR
authors.lastname: Marti
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001202:Ascitic Fluid / Q000166:cytology; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000032:analysis*; D001707:Biopsy, Needle; D016022:Case-Control Studies; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000401:mortality / Q000601:surgery*; D046508:Culture Techniques; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009366:Neoplasm Seeding; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D010533:Peritoneal Lavage; D010865:Pilot Projects; D011379:Prognosis; D016016:Proportional Hazards Models; D011446:Prospective Studies; D012333:RNA, Messenger / Q000032:analysis; D012016:Reference Values; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D018570:Risk Assessment; D012680:Sensitivity and Specificity; D016019:Survival Analysis
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger