title: Preliminary study of immunomagnetic quantification of circulating tumor cells in patients with advanced disease.
abstract: OBJECTIVE
  To enumerate the amount of circulating tumor cells (CTCs) in patients with advanced prostate cancer and to investigate the relationship between these numbers, prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSM) expression, and clinical parameters.
  
  
  METHODS
  Whole blood was collected in proprietary CellSave tubes. Mononuclear cell fractions were isolated using epithelial cell antibody-coated magnetic nanoparticles. On one half of each immunomagnetically enriched cell fraction, automated fluorescent microscopy was used to identify the epithelial tumor cells. From the remainder of each sample, RNA extraction, cDNA synthesis, and polymerase chain reaction amplification of PSA and PSM were performed.
  
  
  RESULTS
  Eighty-four patients with advanced prostate cancer submitted 130 samples for analysis. Intact CTCs were identified in 62% of samples; 83.3% of CTC-positive and 0% of CTC-negative samples were reverse transcriptase-polymerase chain reaction positive for PSA and PSM (P = 0.001). A significant positive correlation was found between the CTC number and PSA (r = 0.49), alkaline phosphatase (r = 0.47), and lactate dehydrogenase (r = 0.55) levels, and a significant negative correlation with hemoglobin (r = -0.35). The initial Gleason grade, prior therapy, current therapy, and type of metastasis (bone, soft tissue) did not correlate significantly with the CTC number.
  
  
  CONCLUSIONS
  The presence of intact CTCs and the expression of PSA and PSM demonstrated robust agreement. The tumor cell numbers reflected current disease status and correlated significantly with the clinical disease indicators of PSA, hemoglobin, and liver function tests. These findings warrant further investigation of the diagnostic and prognostic value of enumerating intact CTCs.
authors.affiliation: Department of Urology, University of Michigan School of Medicine, Ann Arbor, Michigan, USA. btchen@umich.edu
authors.forename: B T
authors.initials: BT
authors.lastname: Chen
authors.forename: R D
authors.initials: RD
authors.lastname: Loberg
authors.forename: C K
authors.initials: CK
authors.lastname: Neeley
authors.forename: S M
authors.initials: SM
authors.lastname: O'Hara
authors.forename: S
authors.initials: S
authors.lastname: Gross
authors.forename: G
authors.initials: G
authors.lastname: Doyle
authors.forename: R L
authors.initials: RL
authors.lastname: Dunn
authors.forename: L M
authors.initials: LM
authors.lastname: Kalikin
authors.forename: K J
authors.initials: KJ
authors.lastname: Pienta
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000954:Antigens, Surface / Q000032:analysis; D018450:Disease Progression; D043425:Glutamate Carboxypeptidase II / Q000032:analysis; D006801:Humans; D018189:Immunomagnetic Separation; D008297:Male; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D017430:Prostate-Specific Antigen / Q000032:analysis; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000473:pathology*
chemical_list: D000954:Antigens, Surface; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II; D017430:Prostate-Specific Antigen