title: Circulating tumor cell analysis in patients with progressive castration-resistant prostate cancer.
abstract: OBJECTIVE
  To better direct targeted therapies to the patients with tumors that express the target, there is an urgent need for blood-based assays that provide expression information on a consistent basis in real time with minimal patient discomfort. We aimed to use immunomagnetic-capture technology to isolate and analyze circulating tumor cells (CTC) from small volumes of peripheral blood of patients with advanced prostate cancer.
  
  
  METHODS
  Blood was collected from 63 patients with metastatic prostate cancer. CTCs were isolated by the Cell Search system, which uses antibodies to epithelial cell adhesion marker and immunomagnetic capture. CTCs were defined as nucleated cells positive for cytokeratins and negative for CD45. Captured cells were analyzed by immunofluorescence, Papanicolau staining, and fluorescence in situ hybridization.
  
  
  RESULTS
  Most patients (65%) had 5 or more CTCs per 7.5 mL blood sample. Cell counts were consistent between laboratories (c = 0.99) and did not change significantly over 72 or 96 h of storage before processing (c = 0.99). Their identity as prostate cancer cells was confirmed by conventional cytologic analysis. Molecular profiling, including analysis of epidermal growth factor receptor (EGFR) expression, chromosome ploidy, and androgen receptor (AR) gene amplification, was possible for all prostate cancer patients with >or=5 CTCs.
  
  
  CONCLUSIONS
  The analysis of cancer-related alterations at the DNA and protein level from CTCs is feasible in a hospital-based clinical laboratory. The alterations observed in EGFR and AR suggest that the methodology may have a role in clinical decision making.
authors.affiliation: Genitourinary Oncology Service, Immunicon Corporation, Huntingdon Valley, Pennsylvania, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Shaffer
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Leversha
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Danila
authors.forename: Oscar
authors.initials: O
authors.lastname: Lin
authors.forename: Rita
authors.initials: R
authors.lastname: Gonzalez-Espinoza
authors.forename: Bin
authors.initials: B
authors.lastname: Gu
authors.forename: Aseem
authors.initials: A
authors.lastname: Anand
authors.forename: Katherine
authors.initials: K
authors.lastname: Smith
authors.forename: Peter
authors.initials: P
authors.lastname: Maslak
authors.forename: Gerald V
authors.initials: GV
authors.lastname: Doyle
authors.forename: Leon W M M
authors.initials: LW
authors.lastname: Terstappen
authors.forename: Hans
authors.initials: H
authors.lastname: Lilja
authors.forename: Glenn
authors.initials: G
authors.lastname: Heller
authors.forename: Martin
authors.initials: M
authors.lastname: Fleisher
authors.forename: Howard I
authors.initials: HI
authors.lastname: Scher
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002369:Castration; D003584:Cytological Techniques; D066246:ErbB Receptors / Q000096:biosynthesis; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005784:Gene Amplification; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D066246:ErbB Receptors; D018719:Erb-b2 Receptor Tyrosine Kinases; D011944:Receptors, Androgen