title: Detection of isolated disseminated tumor cells in bone marrow and blood samples of patients with hepatocellular carcinoma.
abstract: BACKGROUND
  Patients with hepatocellular carcinoma (HCC) often develop recurrences after curative resection or liver transplantation. Therefore, tumor cell dissemination must have occurred preoperatively or intraoperatively. Current staging methods cannot reliably detect micrometastasis. Reverse transcription-polymerase chain reaction (RT-PCR) for alpha-fetoprotein (AFP) has been used to detect circulating liver cancer cells, but results in blood samples have been contradictory.
  
  
  OBJECTIVE
  AFP-RT-PCR is a specific and sensitive assay for the detection of disseminated tumor cells in central venous blood and bone marrow samples of patients with HCC and has prognostic relevance.
  
  
  METHODS
  Prospective consecutive series.
  
  
  METHODS
  University hospital.
  
  
  METHODS
  We performed preoperative, intraoperative, and postoperative analyses of central venous blood samples and preoperative analysis of bone marrow samples of patients with HCC and patients without malignant disease, using a modified AFP-RT-PCR method. Preoperative serum AFP levels were measured. Clinical follow-up ranged from 4 to 20 months.
  
  
  METHODS
  Sensitivity and specificity of AFP-RT-PCR, correlation of AFP-RT-PCR results to tumor stage and tumor recurrence.
  
  
  RESULTS
  In serial dilution experiments, 50 AFP-expressing HepG2 cells were detected in 10 mL of blood. Peripheral blood samples of 20 healthy volunteers and bone marrow samples of 21 patients with benign diseases consistently tested negative for AFP, whereas 4 of 24 patients with HCC showed AFP expression in bone marrow samples. All these patients had advanced disease; however, correlation of positive RT-PCR results to tumor stage was not significant (P = .07). One of the 4 AFP-positive patients developed an intrahepatic recurrence soon after liver transplantation. Central venous blood of patients with HCC (n = 24) and patients with benign liver diseases (n = 13) equally demonstrated AFP-expressing cells. There was no correlation of RT-PCR results to serum AFP levels.
  
  
  CONCLUSIONS
  Perioperative screening for micrometastasis in bone marrow of patients with HCC is sensitive and specific with AFP-RT-PCR and may have prognostic relevance. Alpha-fetoprotein is not a suitable marker for the detection of tumor cells in central venous blood samples.
authors.affiliation: Department of Surgery, University of Heidelberg, Germany. p21@ix.urz.uni-heidelberg.de
authors.forename: P
authors.initials: P
authors.lastname: Kienle
authors.forename: J
authors.initials: J
authors.lastname: Weitz
authors.forename: R
authors.initials: R
authors.lastname: Klaes
authors.forename: M
authors.initials: M
authors.lastname: Koch
authors.forename: A
authors.initials: A
authors.lastname: Benner
authors.forename: T
authors.initials: T
authors.lastname: Lehnert
authors.forename: C
authors.initials: C
authors.lastname: Herfarth
authors.forename: M
authors.initials: M
authors.lastname: von Knebel Doeberitz
mesh_terms: D001853:Bone Marrow / Q000473:pathology; D019046:Bone Marrow Neoplasms / Q000473:pathology / Q000556:secondary*; D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000556:secondary*; D005260:Female; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D000509:alpha-Fetoproteins / Q000032:analysis
chemical_list: D012333:RNA, Messenger; D000509:alpha-Fetoproteins